Banner
Current IssueEditorial BoardOnline First ArticlesArchive
Current IssueEditorial BoardOnline First ArticlesArchive

Full Research Article

Quercetin Verses Metformin, Comparative Efficacy on the Lipid Metabolism Gene Expression in Different Organs of Diabetic Wistar Rats

N
Nayab Khan1,*
0000-0002-3965-6168
A
Asmatullah Kakar1
I
Imtiaz Rabbani2
M
Mahrukh Naseem1
I
Irfan Shahzad Sheikh3
T
Tahir Yaqub4
M
Muhammad Imran5
1Department of Zoology, University of Balochistan Quetta, Pakistan.
2Department of Physiology, University of Veterinary and Animal Sciences, Lahore, Pakistan.
3Center for Advanced Studies in Vaccinology and Biotechnology, University of Balochistan, Quetta, Pakistan.
4Institute of Microbiology, University of Veterinary and Animal Sciences, Lahore, Pakistan.
5Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore, Pakistan.

ABSTRACT

Background: Type 2 Diabetes mellitus, is a metabolic disorder responsible for disturbance in carbohydrate, protein and lipid metabolism. The objective of current study was to evaluate the effect of Quercetin and Metformin on serum lipid profile and gene expression involved in lipid metabolism.

Methods: Sixty-four (64) Wistar rats in a range of live body weight 196.75-198.19 grams, were divided in eight (8) groups (n=8 each); CONTROL, DIABETIC, DIAB+MET, DIAB+25Q, DIAB+50Q, NONDIAB+MET, NONDIAB+25Q and NONDIAB+50Q. A single dose of Streptozotocin drug @ 35 mg per kg was used for the induction of diabetes. The particular doses of Metformin and Quercetin were given by gavage on daily basis in their respective groups till the end of 14-week trial.

Result: In case of FAS gene expression in liver, DIAB+MET (1.26±0.03), DIAB+25Q (1.31±0.02) and DIAB+50Q (1.19±0.04), suppresses the elevated expression compared to Diabetic group, while in case of SREBP-1c, both DIAB+MET (1.31±0.05) and DIAB+50Q (1.38±0.04) showed predominant results compared to DIAB+25Q. Moreover, for PPAR-α, DIAB+MET (0.87±0.06) exhibited considerable upregulation in comparison with DIAB+25Q (0.65±0.03) and DIAB+50Q (0.76±0.04). In adipose, FAS gene was significantly downregulated in DIAB+50Q (1.28±0.03) group as compared to other two diabetic treatment groups, however, for PPAR-γ gene, in term of significance both DIAB+MET (0.84±0.05) and DIAB+50Q (0.88±0.06) displayed statistically similar level of upregulation. In muscles, PPAR-α expression was best observed in DIAB+50Q (0.80±0.03) and DIAB+MET (0.76±0.02) compared to DIAB+25Q (0.59±0.03). Moreover, serum lipid profile (TC, HDL, TG, LDL and VLDL) was also significantly improved in Metformin and Quercetin treatment groups. In non-diabetic treatment groups, a non-significant difference was observed when compared to CONTROL group.

INTRODUCTION

Diabetes, a heterogenous disorder, occurring as a result of environmental and genetic factors, is currently a chronic health issue worldwide (Riddle et al., 2022). Diabetes exists in two major forms: Type 1 Diabetes and Type 2 Diabetes. The type 1 diabetes is basically an autoimmune disorder, involving destruction of pancreatic beta cells due to immune system attacks. This immune response makes the body devoid of insulin, the beta cells hormone accountable for blood glucose regulation. The disease mainly prevails in children but also has a chance of developing in young adult and other age group people. Type 2 diabetes accounting for more than 90% of total diabetes cases (Guo et al., 2023), has a tendency to develop in later stages of life unlike type 1, although now a days owing to the increasing rate of obesity, the disease is also prevailing in young individuals (Olimjonovna, 2024). In this disease type, either the body produce insufficient insulin or becomes insulin resistant (Khajuria et al., 2018; Manikandan et al., 2018; Olimjonovna, 2024), thus failing to manage blood glucose level, which ultimately results in further complications (Devi et al., 2023; Olimjonovna, 2024). The disease is mainly classified by disturbance in fat, carbohydrate and protein metabolism, attributed to dysregulation in insulin action or secretion (Naseem et al., 2020).
       
According to 2017 IDF atlas of diabetes, Pakistan is ranked 2nd among 21 countries of Middle East and North Africa for the prevalence of Diabetes, having nearly 7.5 million individuals affected with the disease aged between 20-79, additionally Pakistan is categorized among 21 countries, to be ranked at position 18 based on 6.9% diabetes prevalence (Adnan and Aasim, 2020). The number is expected to reach 16.7 million in 2045, if proper interventional measured were not adopted by the concerned (Bukhsh et al., 2019).
       
Diabetic dyslipidemia is caused mainly as a result of metabolic disruption and insulin resistance that disturb the lipid metabolism and its related vital enzymes. Diabetes associated dyslipidemia is usually attributed to high serum triglyceride and cholesterol levels, moreover, low levels of HDL-cholesterol. Improving lipid metabolism signifies an important approach towards diabetes treatment. (Rahmani et al., 2023). As plant products are having fewer side effects, interest in anti-hyperglycemic medicines derived from plants has steadily increased in recent years. Now a days, many potential health benefits of polyphenolic compounds, are considered for the treatment of diabetic complications owing to their anti-inflammatory and anti-oxidant properties. Flavonoids (polyphenolic compounds subclass), are known for their beneficial role in lipid metabolism (Umar and Utari, 2019), which is often affected due to prolong untreated diabetes (Kane et al., 2021). Flavonoid also exhibit varied beneficial health effects for diabetes management, i.e., improving insulin sensitivity and secretion along with slowing carbohydrates absorption in intestine. One of the vital flavonoids considered for treating diabetic complications (including dyslipidemia) is Quercetin. A considerable number of research studies are available, focusing on antidiabetic properties of Quercetin and highlighting their importance in relieving diabetic complications (Yan et al., 2023). Quercetin ameliorate lipid associated metabolism and liver injury, moreover its protective effect is also evident from its role in mitigating diabetes related cardiovascular injury (Hosseini et al., 2021), for which the major risk factor being considered is dyslipidemia (Arvanitis and Lowenstein, 2023). The vital regulators of lipid metabolism include genes namely FAS, PPAR-α, SREBP-1c and PPAR-γ, therefore they are used as molecular marker for various metabolic syndrome especially dyslipidemia (Karimi-Sales et al., 2019).
               
To our best knowledge, there is not even a single study pertaining to comparative anti-dyslipidemia effect of organic product Quercetin and synthetic drug Metformin. The main objective of current research was to compare the anti-dyslipidemia effect of Quercetin and Metformin, in term of gene expression and lipid profile.

MATERIALS AND METHODS

Animal acclimatation
 
For this particular research, Wistar rats were kept in animal shed of University of Balochistan (Quetta, Pakistan), providing appropriate acclimatization period. To obtain homogenized live body weights, all the rats had free access to food and water, controlled environment with optimal temperature 24±5oC and 12-12 hour of light dark cycle each. Prior trial, experimental rats were allowed to feed on high fat diet (6% cellulose, 12.8% maize starch, 6.6% dextrose, 28.6% casein, 31.1% beef tallow, 9.7% minerals, 1.3% vitamins and 3.9% sun flower oil) for 21 days. The ethical approval for conducting this research was obtained from ethical committee, University of Balochistan.
 
Animal grouping
 
• At week 0, sixty-four (64) Wistar rats in a range of live body weight 196.75-198.19 grams, were divided in eight (8) groups (n=8 each). A single dose of Streptozotocin drug (MS07936, Biosynth, Switzerland) @ 35 mg per kg was used for the induction of diabetes. The particular doses of Metformin (Glucophage, Martindow, Pakistan) and Quercetin (Quercetin hydrate, 95%, Thermo Scientific Chemicals, US.) were given by gavage on daily basis in their respective groups till the end of 14-week trial (Naseem et al., 2020).
 
♦ CONTROL group
 
This group comprised of normal rats (negative control) and were fed on normal diet.
 
♦ DIABETIC group
 
This group was positive control and consisted of diabetic rats that were fed on normal diet.
 
♦ DIAB+MET (Diabetic plus 100 mg/kg metformin)
 
In this group, rats were diabetic and were kept on 100 mg antidiabetic metformin drug.
 
♦ DIAB+25Q (Diabetic plus 25 mg Quercetin)
 
Here, the rats were also diabetic and were provided with 25 mg Quercetin per kg per day.
 
♦ DIAB+50Q (Diabetic plus 50 mg Quercetin)
 
This group comprised of diabetic rats, kept on 50 mg Quercetin.
 
♦NONDIAB+MET (Non-Diabetic plus 100 mg metformin)
 
This experimental group included rats that were non- diabetic and were provided with 100 mg metformin drug.
 
♦ NONDIAB+25Q (Non-Diabetic plus 25 mg Quercetin)
 
This group comprised of non-diabetic rats, kept on 25 mg Quercetin per kg per day.
 
♦ NONDIAB+50Q (Non-Diabetic plus 50 mg Quercetin)
 
This non-diabetic group was given 50 mg Quercetin.
 
Extraction of serum
 
After 14 weeks trial period, blood was collected in vacutainer serum gel collection tubes. Following clotting of blood, these tubes were centrifuged at 2500 rpm for 10 minutes to collect serum, that was stored at -80oC for lipid profiling through micro lab-300 using Innoline commercial kits.

mRNA extraction, cDNA synthesis, real time PCR
 
The skeletal muscle, hepatic and adipose tissues were isolated from rats, washed with saline water and were stored at -80oC. Following the manufacturer’s instructions, Trizol reagent (Bioshop, Canada) was used for the extraction of RNA from the frozen samples of all the three tissues. The commercial RNA extraction kit (CO2203403, Ascend.bio, UK) was used. The extracted RNA was quantified using nanodrop (absorbance at 260 nm). RNA (1 µg) was changed into cDNA by the process of reverse transcription in a reaction volume of total 20 µl using cDNA kit (01317155, Thermo-scientific, Lithuania). The conventional  PCR was used for the conversion of RNA into cDNA, involving annealing at 25oC for 5 min, elongation step at 42oC for 60 min and enzyme inactivation at 70oC for 5 min. Quantitative PCR with the aid of SYBR Green master mix (Thermos-scientific) was operated on a Qiagen (Germany) following the instructions given by the manufacturer. The PCR was performed for a total of 45 cycles. The PCR product was monitored continuously by reading the fluorescence throughout the reaction. The gene expression results were analyzed on Rotor gene Q series software. Primer3 website was used for designing the primers. The GAPDH (housekeeping gene) was used for normalizing the mRNA levels. The delta Ct method was used for the relative quantification. The primers used in real time PCR are mentioned in Table 1.

Table 1: Lipid metabolism genes and their primer sequences.


 
Statistical analysis
 
Results of the current study are represented as mean ± SEM. One way ANOVA was used for the analysis followed by Duncan multiple range test. Results were analyzed at level of significance (p<0.05), which was considered as significant for the particular study.

RESULTS AND DISCUSSION

Diabetes is regarded as one of the major health concerns worldwide. More than 90% of diabetes cases are attributed to type 2 diabetes mellitus and this particular disease is often ascribed as multifactorial origin chronic metabolic disorder. Diabetes is responsible for disturbing the lipid metabolism and causing inflammation in body. Polyphenols are phytochemicals that are present in vegetables and fruits, moreover they are further categorized into flavonoids, phenolic acids, stilbenes and lignans. Foods rich in Polyphenol and dietary supplements both are linked with a lower occurrence of lipid dependent cardiovascular disease (Costa et al., 2022). A common flavonoid, Quercetin, readily available in vegetables and fruits, is a potent antioxidant. Quercetin exhibit varied beneficial features, namely anti-diabetic, anti-dyslipidemia, anti-inflammatory, anti-cancer and antioxidant properties (Wang et al., 2022). Metformin hydrochloride with its anti-hyperglycemic and anti-dyslipidemia ability is also being in practice for diabetes treatment (Pournaghi et al., 2012; Szymczak-Pajor et al., 2022). Quercetin and Metformin play a crucial role in lipid metabolism through lipid metabolism gene expression. In our study, lipid profiling was done at the end of trial. Gene expression data was also recorded in liver, adipose tissue and skeletal muscles individually at the end of 14 weeks trial after slaughtering the rats.
 
Ameliorative effect of quercetin and metformin on lipid profile in wistar rats
 
In blood serum, lipid levels are good indicators of cardiovascular health. It is a well-documented fact that untreated type 2 diabetes mellitus results in elevated levels of LDL, triglycerides, VLDL and also decreased level of HDL, promoting progression to coronary artery diseases. Moreover, the deficiency of insulin in diabetes leads to a variety of irrationalities in regulatory and metabolic pathways, which ultimately results in accumulation of lipids namely TC and triglycerides in diabetic individuals. At the end of 14-week trial, serum was isolated from all the 64 albino rats for biochemical analysis. The results were revealed for serum TC, HDL, TG, LDL-C and VLDL-C (Table 2).

Table 2: Lipid profile of Wistar rats in different treatment groups.


       
In our research study, streptozotocin induced diabetic group displayed a considerable increase in LDL, VLDL, triglyceride, total cholesterol while a significant decrease in HDL levels as compared to CONTROL group rats. Following 14 week treatment, both Quercetin treated and Metformin treated diabetic rat groups showed a substantial reduction in LDL, VLDL, triglyceride, total cholesterol and elevated levels of HDL (Table 2). Earlier researches have also found improvement in lipid profile following Quercetin and Metformin treatment (Albasher et al., 2020; Ali et al., 2020; Abdelkader et al., 2020; Lai et al., 2021; Hacioglu et al., 2021; Chellian et al., 2022; Hu et al., 2022; Jiang et al., 2022; Rahmani et al., 2023).

Gene expression of FAS, SREBP-1c and PPAR-α in liver
 
In liver, FAS gene expression showed a significant difference (p<0.05) between CONTROL (1.03±0.09) and DIABETIC group (1.55±0.03), While other groups i.e. DIAB+MET (1.26±0.03), DIAB+25Q (1.31±0.02) and DIAB+50Q (1.19±0.04) showed a considerable difference (p<0.05) compared to DIABETIC group. In term of significance, all the three treatment groups (DIAB+MET, DIAB+25Q and DIAB+50Q) displayed statistically similar downregulation of FAS gene. There was a significant difference (p<0.05) between CONTROL and treatment groups (DIAB+MET, DIAB+25Q and DIAB+50Q). A non-significant difference was observed between CONTROL (1.03±0.09) and non-diabetic treatment groups; NONDIAB +MET (1.01±0.03), NONDIAB+25Q (1.03±0.05) and NONDIAB+50Q (0.98±0.04) (Fig 1).

Fig 1: Gene expression of FAS, SREBP-1c and PPARa in liver.


       
SREBP1-c gene expression in liver, also displayed a substantial difference (p<0.05) between CONTROL (1.05±0.12) and DIABETIC group (2.33±0.03), Whereas the treatment groups with diabetes i.e. DIAB+MET (1.31±0.05), DIAB+25Q (1.70±0.02) and DIAB+50Q (1.38±0.04) showed a considerable difference (p<0.05) compared to DIABETIC group. The DIAB+MET and DIAB+50Q groups exhibited major reduction in expression of SREBP1-c gene compared to DIAB+25Q. There was a significant difference (p<0.05) between CONTROL and treatment groups (DIAB+MET, DIAB+25Q and DIAB+50Q). A non-significant difference was observed between CONTROL (1.05±0.12) and non-diabetic treatment groups; NONDIAB+MET (0.96±0.02), NONDIAB+25Q (1.01±0.04) and NONDIAB+50Q  (0.98±0.04) (Fig 1).
       
PPAR-α gene expression in liver, revealed a significant difference (p<0.05) between CONTROL (1.00±0.02) and DIABETIC group (0.54±0.04). The diabetic treatment groups i.e. DIAB+MET (0.87±0.06), DIAB+25Q (0.65±0.03) and DIAB+50Q (0.76±0.04) showed a considerable difference (p<0.05) compared to DIABETIC group, showing an improvement in liver PPAR-α gene expression compared to DIABETIC. The DIAB+MET group showed relatively better expression of PPAR-α gene compared to the other two diabetic treatment groups. A non-significant difference was observed between CONTROL (1.00±0.02) and non-diabetic treatment groups; NONDIAB+MET (1.00±0.04), NONDIAB+25Q (1.00±0.03) and NONDIAB+50Q (1.02±0.03) (Fig 1).
 
Gene expression of FAS and PPAR-γ in adipose tissue
 
In adipose tissue, FAS gene expression showed a significant variation (p<0.05) between CONTROL (1.01±0.06) and DIABETIC group (2.13±0.05), While other groups i.e. DIAB+MET (1.40±0.04), DIAB+25Q (1.72±0.04) and DIAB+50Q (1.28±0.03) showed a significant difference (p<0.05) compared to DIABETIC group. The most considerable downregulation of FAS gene was observed in DIAB+50Q group. There was a significant difference (p<0.05) between CONTROL and treatment groups (DIAB+MET, DIAB+25Q and DIAB+50Q). A non-significant difference was observed between CONTROL (1.01±0.06) and non-diabetic treatment groups; NONDIAB+MET (0.99±0.02), NONDIAB+25Q (1.01±0.03) and NONDIAB+50Q (0.97±0.03) (Fig 2).

Fig 2: Gene expression of FAS and PPAR g in Adipose tissue.


       
PPAR-γ gene expression in adipose, displayed a substantial difference (p<0.05) between CONTROL (1.11±0.18) and DIABETIC group (0.33±0.05), Whereas the treatment groups with diabetes i.e. DIAB+MET (0.84±0.05), DIAB+25Q (0.61±0.03) and DIAB+50Q (0.88±0.06) showed a significant difference (p<0.05) compared to DIABETIC group. A non-significant difference was observed between CONTROL (1.11±0.18) and non-diabetic treatment groups; NONDIAB+MET (1.15±0.04), NONDIAB+25Q (1.12±0.05) and NONDIAB+50Q (1.17±0.06) (Fig 2).
 
Gene expression of PPAR-α in skeletal muscles
 
PPAR-α gene expression in skeletal muscle, showed a significant variance (p<0.05) between CONTROL (1.00±0.03) and DIABETIC group (0.39±0.02). The diabetic treatment groups i.e. DIAB+MET (0.76±0.02), DIAB+25Q (0.59±0.03) and DIAB+50Q (0.80±0.03) showed a substantial difference (p<0.05) compared to DIABETIC group, indicating an improvement in skeletal muscle PPAR-α gene expression compared to DIABETIC. The DIAB+MET and DIAB+50Q groups showed similar upregulation of PPAR-α gene, with no significant difference between the two groups. A non-significant difference was observed between CONTROL (1.00±0.03) and non-diabetic treatment groups; NONDIAB +MET (1.01±0.02), NONDIAB+25Q (0.98±0.03) and NONDIAB+50Q (1.03±0.03) (Fig 3).

Fig 3: Gene expression of PPAR-a in skeletal muscles.


       
The expression data of PPAR-α in our study is similar with previously reported studies (Karimi-Sales et al., 2019; Malinska et al., 2019; Ting et al., 2018). PPAR-α is disseminated in several body tissues but is mainly present in liver, adipose and skeletal tissues. PPAR-α is involved in gene regulation specifically genes concerned with lipoprotein metabolism, lipid transport and fatty acid oxidation (Małodobra-Mazur et al., 2024).
       
In term of PPAR-γ expression, the findings for Quercetin (Sun et al., 2015) are in line with previous studies. The experimental outcome of our study for PPAR-γ are also in accordance with previous study reporting an upregulation mediated by flavonoid  (Singh et al., 2018). Matsukawa et al., 2015, also found a significant upregulation of PPAR-γ in diabetic mice after treatment of mice with flavonoid, which is in accordance with our study results. PPAR-γ is regarded as the adipogenesis master regulator, as it increases the storage of lipid. PPAR-δ is also responsible for regulation of the change from glycolytic state to oxidative muscle fibres (Montaigne et al., 2021).
       
SREBPs role in lipid metabolism is multifunctional and thereby numerous metabolic issues are related to dysregulation of SREBP for example dyslipidemia, type 2 diabetes, atherosclerosis and fatty liver disease (Li et al., 2023). SREBP-1c is a vital protein responsible for synthesizing fat and when SREBP-1c is activated then it further enhances adipogenic genes transcription such as fatty acid synthase and acetyl-CoA carboxylase, thus accelerating hepatic fat production (Gan et al., 2024). In case of gene expression involving SREBP-1c, Lee and Kim (2022) also found results relevant with out findings, reporting a down regulation of SREBP-1c in Quercetin treated groups in high fat diet model. Pengnet et al. (2022), also presented a relatable result, showing a downregulation of SREBP-1c in rats after flavonoid treatment. Karimi-Sales et al. (2019), presented a similar result, a downregulation of SREBP-1c in wistar rats following flavonoid treatment which is a similar trend as in our study.
       
Our study outcome for FAS expression is in accordance with previously conducted studies (Afarin et al., 2024; Lee and Kim 2022). Fatty acid is synthesized by FAS in liver (Sahin et al., 2021) and other tissues namely adipose and skeletal muscle. In diabetes, there is an evident rise in FAS expression and thus consequently leading to dyslipidemia, as FAS is involved in lipid synthesis (Fang et al., 2024). In our study, diabetes induction also causes upregulation of FAS at a considerable level thereby affecting lipid metabolism while treatment with Quercetin and Metformin downregulated the FAS gene.

CONCLUSION

Our study outcome clearly showed that Quercetin flavonoid is equally effective in ameliorating lipid profile and its related gene expression as Metformin which is a synthetic drug. Moreover, the higher dose of 50 mg Quercetin showed comparatively better results in term of lipid profile and gene expression as compared to 25 mg dose.

CONFLICT OF INTEREST

All authors declared that there is no conflict of interest.

REFERENCES


  1. Abdelkader, N.F., Eitah, H.E., Maklad, Y.A., Gamaleldin, A.A., Badawi, M.A. and Kenawy, S.A. (2020). New combination therapy of gliclazide and quercetin for protection against STZ- induced diabetic rats. Life Sciences. 247: 117458.

  2. Adnan, M. and Aasim, M. (2020). Prevalence of type 2 diabetes mellitus in adult population of Pakistan: A meta-analysis of prospective cross-sectional surveys. Annals of Global Health. 86(1).

  3. Afarin, R., Hatami, M., Monjezi, S., Bineshfar, F. and Ahangarpour, A. (2024). Suppression of TGF-β/Smad3 signaling pathway by Capparis spinosa and quercetin in a rat model of nonalcoholic steatohepatitis. Iranian Journal of Basic Medical Sciences. 27(9): 1096.

  4. Albasher, G., Alwahaibi, M., Abdel-Daim, M.M., Alkahtani, S. and Almeer, R. (2020). Protective effects of Artemisia judaica extract compared to metformin against hepatorenal injury in high-fat diet/streptozotocine-induced diabetic rats. Environmental Science and Pollution Research. 27: 40525-40536.

  5. Ali, A.M., Gabbar, M.A., Abdel-Twab, S.M., Fahmy, E.M., Ebaid, H., Alhazza, I.M. and Ahmed, O.M. (2020). Antidiabetic potency, antioxidant effects and mode of actions of Citrus reticulata fruit Peel hydroethanolic extract, hesperidin and quercetin in nicotinamide/streptozotocin induced wistar diabetic rats. Oxidative Medicine and Cellular Longevity. 2020(1): 1730492.

  6. Arvanitis, M. and Lowenstein, C.J. (2023). Dyslipidemia. Annals of Internal Medicine. 176(6): ITC81-ITC96.

  7. Bukhsh, A., Khan, T.M., Sarfraz Nawaz, M., Sajjad Ahmed, H., Chan, K.G. and Goh, B.H. (2019). Association of diabetes knowledge with glycemic control and self-care practices among Pakistani people with type 2 diabetes mellitus. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy. pp: 1409-1417.

  8. Chellian, J., Mak, K.K., Chellappan, D.K., Krishnappa, P. and Pichika, M.R. (2022). Quercetin and metformin synergistically reverse endothelial dysfunction in the isolated aorta of streptozotocin-nicotinamide-induceddiabetic rats. Scientific Reports. 12(1): 21393.

  9. Costa, P.C.T.D., de Souza, E.L., Lacerda, D.C., Cruz Neto, J.P.R., Sales, L.C.S.D., Silva Luis, C.C. and de Brito Alves, J.L. (2022). Evidence for quercetin as a dietary supplement for the treatment of cardio-metabolic diseases in pregnancy: A review in rodent models. Foods. 11(18): 2772.

  10. Devi, G., Dimri, U., Ajith, Y., Deepa, P.M., Yatoo, M.I., Gopalakrishnan, A. and Madhesh, E. (2023). The anti-oxidant and the anti-diabetic effects of terminalia chebula and withania somnifera in subclinically diabetic dogs. Indian Journal of Animal Research. 57(8): 1042-1050. doi: 10.18805/ IJAR.B-4355.

  11. Fang, Z., Liu, R., Xie, J. and He, J.C. (2024). Molecular mechanism of renal lipid accumulation in diabetic kidney disease. Journal of Cellular and Molecular Medicine. 28(11): e18364.

  12. Gan, L., Guo, H., Yang, Q., Zhou, X., Xie, Y., Ma, X. and Zuo, Z. (2024). Alkaline mineral complex water attenuates transportation-induced hepatic lipid metabolism dysregulation by AMPKα-SREBP-1c/PPARα pathways. International Journal of Molecular Sciences. 25(21): 11373.

  13. Guo, H., Wu, H. and Li, Z. (2023). The pathogenesis of diabetes. International Journal of Molecular Sciences. 24(8): 6978.

  14. Hacioglu, C., Kar, F., Kara, Y., Yucel, E., Donmez, D.B., Sentürk, H. and Kanbak, G. (2021). Comparative effects of metformin and Cistus laurifolius L. extract in streptozotocin-induced diabetic rat model: Oxidative, inflammatory, apoptotic and histopathological analyzes. Environmental Science and Pollution Research. 28(41): 57888-57901.

  15. Hosseini, A., Razavi, B.M., Banach, M. and Hosseinzadeh, H. (2021). Quercetin and metabolic syndrome: A review. Phytotherapy  Research. 35(10): 5352-5364.

  16. Hu, N., Zhang, Q., Wang, H., Yang, X., Jiang, Y., Chen, R. and Wang,  L. (2022). Comparative evaluation of the effect of metformin and insulin on gut microbiota and metabolome profiles of type 2 diabetic rats induced by the combination of streptozotocin and high-fat diet. Frontiers in Pharmacology. 12: 794103.

  17. Jiang, J.J., Zhang, G.F., Zheng, J.Y., Sun, J.H. and Ding, S.B. (2022). Targeting mitochondrial ROS-mediated ferroptosis by quercetin alleviates high-fat diet-induced hepatic lipotoxicity. Frontiers in Pharmacology. 13: 876550.

  18. Kane, J.P., Pullinger, C.R., Goldfine, I.D. and Malloy, M.J. (2021). Dyslipidemia and diabetes mellitus: Role of lipoprotein species and interrelated pathways of lipid metabolism in diabetes mellitus. Current Opinion in Pharmacology. 61: 21-27.

  19. Karimi-Sales, E., Ebrahimi-Kalan, A. and Alipour, M.R. (2019). Preventive effect of trans-chalcone on non-alcoholic steatohepatitis: Improvement of hepatic lipid metabolism. Biomedicine and Pharmacotherapy. 109: 1306-1312.

  20. Khajuria, P., Raghuwanshi, P., Rastogi, A., Koul, A.L., Zargar, R.  and Kour, S. (2018). Hepatoprotective effect of Seabuckthorn leaf extract in streptozotocin induced diabetes mellitus in Wistar rats. Indian Journal of Animal Research. 52(12): 1745-1750. doi: 10.18805/ijar.B-3439.

  21. Lai, L.L., Lu, H.Q., Li, W.N., Huang, H.P., Zhou, H.Y., Leng, E.N. and Zhang, Y.Y. (2021). Protective effects of quercetin and crocin in the kidneys and liver of obese sprague-dawley rats with Type 2 diabetes: Effects of quercetin and crocin on T2DM rats. Human and Experimental Toxicology. 40(4): 661-672.

  22. Lee, M.S. and Kim, Y. (2022). Effects of quercetin on the improvement of lipid metabolism through regulating hepatic AMPK and microRNA-21 in high cholesterol diet-fed mice. Journal of Nutrition and Health. 55(1): 36-46.

  23. Li, N., Li, X., Ding, Y., Liu, X., Diggle, K., Kisseleva, T. and Brenner, D.A. (2023). SREBP regulation of lipid metabolism in liver disease and therapeutic strategies. Biomedicines. 11(12): 3280.

  24. Malinska, H., Hüttl, M., Oliyarnyk, O., Markova, I., Poruba, M., Racova, Z. and Vecera, R. (2019). Beneficial effects of troxerutin on metabolic disorders in non-obese model of metabolic syndrome. Plos One. 14(8): e0220377.

  25. Małodobra-Mazur, M., O³dakowska, M. and Dobosz, T. (2024). Exploring PPAR gamma and PPAR alpha’s regulation role in metabolism via epigenetics mechanism. Biomolecules. 14(11): 1445.

  26. Manikandan, R., Anand, A.V., Sampathkumar, P. and Manoharan, N. (2018). Protective effect of Psidium guajava leaf ethanolic extract against streptozotocin-induced diabetes and lipidosis in rats. Indian Journal of Animal Research. 52(8): 1198-1205. doi: 10.18805/ijar.B-3337.

  27. Matsukawa, T., Inaguma, T., Han, J., Villareal, M.O. and Isoda, H. (2015). Cyanidin-3-glucoside derived from black soybeans ameliorate type 2 diabetes through the induction of differentiation of preadipocytes into smaller and insulin- sensitive adipocytes. The Journal of Nutritional Biochemistry. 26(8): 860-867.

  28. Montaigne, D., Butruille, L. and Staels, B. (2021). PPAR control of metabolism and cardiovascular functions. Nature Reviews Cardiology. 18(12): 809-823.

  29. Naseem, M., Ouguerram, K., Nazih, H., Rabbani, I., Zaneb, H., Rehman, H.U. and Tahir, S.K. (2020). Synergistic effect of extracts of Ginkgo biloba leaf and Panax ginseng root on carbohydrate and lipid metabolism gene expression in alloxan induced diabetic rats. JAPS: Journal of Animal and Plant Sciences. 30(6).

  30. Olimjonovna, K.O. (2024). Understanding the causes and risk factors of diabetes. Biologiya Va Kimyo Fanlari Ilmiy Jurnali. 2(5): 1-7.

  31. Pengnet, S., Sumarithum, P., Phongnu, N., Prommaouan, S., Kantip, N., Phoungpetchara, I. and Malakul, W. (2022). Naringin attenuates fructose-induced NAFLD progression in rats through reducing endogenous triglyceride synthesis and activating the Nrf2/HO-1 pathway. Frontiers in Pharma- cology. 13: 1049818.

  32. Pournaghi, P., Sadrkhanlou, R.A., Hasanzadeh, S. and Foroughi, A. (2012). An investigation on body weights, blood glucose levels and pituitary-gonadal axis hormones in diabetic and metformin-treated diabetic female rats. In Veterinary Research Forum. 3(2): 79. 

  33. Rahmani, A.H., Alsahli, M.A., Khan, A.A. and Almatroodi, S.A. (2023). Quercetin, a plant flavonol attenuates diabetic complications, renal tissue damage, renal oxidative stress and inflammation in streptozotocin-induced diabetic rats. Metabolites. 13(1): 130.

  34. Riddle, M.C., Cefalu, W.T., Evans, P.H., Gerstein, H.C., Nauck, M.A., Oh, W.K. and Twenefour, D. (2022). Consensus report: definition and interpretation of remission in type 2 diabetes. The Journal of Clinical Endocrinology and Metabolism. 107(1): 1-9.

  35. Sahin, K., Orhan, C., Kucuk, O., Erten, F., Tuzcu, M., Sahin, N. and Komorowski, J.R. (2021). Effects of magnesium biotinate supplementation on serum insulin, glucose and lipid parameters along with liver protein levels of lipid metabolism in rats. Magnesium Research. 34(1): 9-19.

  36. Singh, A.K., Raj, V., Keshari, A.K., Rai, A., Kumar, P., Rawat, A. and Saha, S. (2018). Isolated mangiferin and naringenin exert antidiabetic effect via PPARγ/GLUT4 dual agonistic action with strong metabolic regulation. Chemico-Biological Interactions. 280: 33-44.

  37. Sun, L., Li, E., Wang, F., Wang, T., Qin, Z., Niu, S. and Qiu, C. (2015). Quercetin increases macrophage cholesterol efflux to inhibit foam cell formation through activating PPARγ- ABCA1 pathway. International Journal of Clinical and Experimental Pathology. 8(9): 10854.

  38. Szymczak-Pajor, I., Wenclewska, S. and Śliwińska, A. (2022). Metabolic action of metformin. Pharmaceuticals. 15(7): 810.

  39. Ting, Y., Chang, W.T., Shiau, D.K., Chou, P.H., Wu, M.F. and Hsu, C.L. (2018). Antiobesity efficacy of quercetin-rich supplement on diet-induced obese rats: Effects on body composition, serum lipid profile and gene expression. Journal of Agricultural and Food Chemistry. 66(1): 70-80.

  40. Umar, H.B. and Utari, D.M. (2019). Differences in consumption of flavonoid phytochemicals toward total cholesterol and LDL-cholesterol levels in dyslipidemia and non-dyslipidemia groups of minangkabau ethnic women. Indian Journal of Public Health Research and Development. 10(12).

  41. Wang, D., Li, Y., Zhai, Q.Q., Zhu, Y.F., Liu, B.Y. and Xu, Y. (2022). Quercetin ameliorates testosterone secretion disorder by inhibiting endoplasmic reticulum stress through the miR-1306-5p/HSD17B7 axis in diabetic rats. Bosnian Journal of Basic Medical Sciences. 22(2): 191. 

  42. Yan, L., Vaghari-Tabari, M., Malakoti, F., Moein, S., Qujeq, D., Yousefi, B. and Asemi, Z. (2023). Quercetin: An effective polyphenol in alleviating diabetes and diabetic complications. Critical Reviews in Food Science and Nutrition. 63(28): 9163- 9186. 
Published In
Indian Journal of Animal Research
Article Metrics
Metrics Icon
0
Views
0
Citations
Reviewed By
Share Article
Download Article
In this Article
    Contact us
    Follow us

    Full Research Article

    Quercetin Verses Metformin, Comparative Efficacy on the Lipid Metabolism Gene Expression in Different Organs of Diabetic Wistar Rats

    N
    Nayab Khan1,*
    0000-0002-3965-6168
    A
    Asmatullah Kakar1
    I
    Imtiaz Rabbani2
    M
    Mahrukh Naseem1
    I
    Irfan Shahzad Sheikh3
    T
    Tahir Yaqub4
    M
    Muhammad Imran5
    1Department of Zoology, University of Balochistan Quetta, Pakistan.
    2Department of Physiology, University of Veterinary and Animal Sciences, Lahore, Pakistan.
    3Center for Advanced Studies in Vaccinology and Biotechnology, University of Balochistan, Quetta, Pakistan.
    4Institute of Microbiology, University of Veterinary and Animal Sciences, Lahore, Pakistan.
    5Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore, Pakistan.

    ABSTRACT

    Background: Type 2 Diabetes mellitus, is a metabolic disorder responsible for disturbance in carbohydrate, protein and lipid metabolism. The objective of current study was to evaluate the effect of Quercetin and Metformin on serum lipid profile and gene expression involved in lipid metabolism.

    Methods: Sixty-four (64) Wistar rats in a range of live body weight 196.75-198.19 grams, were divided in eight (8) groups (n=8 each); CONTROL, DIABETIC, DIAB+MET, DIAB+25Q, DIAB+50Q, NONDIAB+MET, NONDIAB+25Q and NONDIAB+50Q. A single dose of Streptozotocin drug @ 35 mg per kg was used for the induction of diabetes. The particular doses of Metformin and Quercetin were given by gavage on daily basis in their respective groups till the end of 14-week trial.

    Result: In case of FAS gene expression in liver, DIAB+MET (1.26±0.03), DIAB+25Q (1.31±0.02) and DIAB+50Q (1.19±0.04), suppresses the elevated expression compared to Diabetic group, while in case of SREBP-1c, both DIAB+MET (1.31±0.05) and DIAB+50Q (1.38±0.04) showed predominant results compared to DIAB+25Q. Moreover, for PPAR-α, DIAB+MET (0.87±0.06) exhibited considerable upregulation in comparison with DIAB+25Q (0.65±0.03) and DIAB+50Q (0.76±0.04). In adipose, FAS gene was significantly downregulated in DIAB+50Q (1.28±0.03) group as compared to other two diabetic treatment groups, however, for PPAR-γ gene, in term of significance both DIAB+MET (0.84±0.05) and DIAB+50Q (0.88±0.06) displayed statistically similar level of upregulation. In muscles, PPAR-α expression was best observed in DIAB+50Q (0.80±0.03) and DIAB+MET (0.76±0.02) compared to DIAB+25Q (0.59±0.03). Moreover, serum lipid profile (TC, HDL, TG, LDL and VLDL) was also significantly improved in Metformin and Quercetin treatment groups. In non-diabetic treatment groups, a non-significant difference was observed when compared to CONTROL group.

    INTRODUCTION

    Diabetes, a heterogenous disorder, occurring as a result of environmental and genetic factors, is currently a chronic health issue worldwide (Riddle et al., 2022). Diabetes exists in two major forms: Type 1 Diabetes and Type 2 Diabetes. The type 1 diabetes is basically an autoimmune disorder, involving destruction of pancreatic beta cells due to immune system attacks. This immune response makes the body devoid of insulin, the beta cells hormone accountable for blood glucose regulation. The disease mainly prevails in children but also has a chance of developing in young adult and other age group people. Type 2 diabetes accounting for more than 90% of total diabetes cases (Guo et al., 2023), has a tendency to develop in later stages of life unlike type 1, although now a days owing to the increasing rate of obesity, the disease is also prevailing in young individuals (Olimjonovna, 2024). In this disease type, either the body produce insufficient insulin or becomes insulin resistant (Khajuria et al., 2018; Manikandan et al., 2018; Olimjonovna, 2024), thus failing to manage blood glucose level, which ultimately results in further complications (Devi et al., 2023; Olimjonovna, 2024). The disease is mainly classified by disturbance in fat, carbohydrate and protein metabolism, attributed to dysregulation in insulin action or secretion (Naseem et al., 2020).
           
    According to 2017 IDF atlas of diabetes, Pakistan is ranked 2nd among 21 countries of Middle East and North Africa for the prevalence of Diabetes, having nearly 7.5 million individuals affected with the disease aged between 20-79, additionally Pakistan is categorized among 21 countries, to be ranked at position 18 based on 6.9% diabetes prevalence (Adnan and Aasim, 2020). The number is expected to reach 16.7 million in 2045, if proper interventional measured were not adopted by the concerned (Bukhsh et al., 2019).
           
    Diabetic dyslipidemia is caused mainly as a result of metabolic disruption and insulin resistance that disturb the lipid metabolism and its related vital enzymes. Diabetes associated dyslipidemia is usually attributed to high serum triglyceride and cholesterol levels, moreover, low levels of HDL-cholesterol. Improving lipid metabolism signifies an important approach towards diabetes treatment. (Rahmani et al., 2023). As plant products are having fewer side effects, interest in anti-hyperglycemic medicines derived from plants has steadily increased in recent years. Now a days, many potential health benefits of polyphenolic compounds, are considered for the treatment of diabetic complications owing to their anti-inflammatory and anti-oxidant properties. Flavonoids (polyphenolic compounds subclass), are known for their beneficial role in lipid metabolism (Umar and Utari, 2019), which is often affected due to prolong untreated diabetes (Kane et al., 2021). Flavonoid also exhibit varied beneficial health effects for diabetes management, i.e., improving insulin sensitivity and secretion along with slowing carbohydrates absorption in intestine. One of the vital flavonoids considered for treating diabetic complications (including dyslipidemia) is Quercetin. A considerable number of research studies are available, focusing on antidiabetic properties of Quercetin and highlighting their importance in relieving diabetic complications (Yan et al., 2023). Quercetin ameliorate lipid associated metabolism and liver injury, moreover its protective effect is also evident from its role in mitigating diabetes related cardiovascular injury (Hosseini et al., 2021), for which the major risk factor being considered is dyslipidemia (Arvanitis and Lowenstein, 2023). The vital regulators of lipid metabolism include genes namely FAS, PPAR-α, SREBP-1c and PPAR-γ, therefore they are used as molecular marker for various metabolic syndrome especially dyslipidemia (Karimi-Sales et al., 2019).
                   
    To our best knowledge, there is not even a single study pertaining to comparative anti-dyslipidemia effect of organic product Quercetin and synthetic drug Metformin. The main objective of current research was to compare the anti-dyslipidemia effect of Quercetin and Metformin, in term of gene expression and lipid profile.

    MATERIALS AND METHODS

    Animal acclimatation
     
    For this particular research, Wistar rats were kept in animal shed of University of Balochistan (Quetta, Pakistan), providing appropriate acclimatization period. To obtain homogenized live body weights, all the rats had free access to food and water, controlled environment with optimal temperature 24±5oC and 12-12 hour of light dark cycle each. Prior trial, experimental rats were allowed to feed on high fat diet (6% cellulose, 12.8% maize starch, 6.6% dextrose, 28.6% casein, 31.1% beef tallow, 9.7% minerals, 1.3% vitamins and 3.9% sun flower oil) for 21 days. The ethical approval for conducting this research was obtained from ethical committee, University of Balochistan.
     
    Animal grouping
     
    • At week 0, sixty-four (64) Wistar rats in a range of live body weight 196.75-198.19 grams, were divided in eight (8) groups (n=8 each). A single dose of Streptozotocin drug (MS07936, Biosynth, Switzerland) @ 35 mg per kg was used for the induction of diabetes. The particular doses of Metformin (Glucophage, Martindow, Pakistan) and Quercetin (Quercetin hydrate, 95%, Thermo Scientific Chemicals, US.) were given by gavage on daily basis in their respective groups till the end of 14-week trial (Naseem et al., 2020).
     
    ♦ CONTROL group
     
    This group comprised of normal rats (negative control) and were fed on normal diet.
     
    ♦ DIABETIC group
     
    This group was positive control and consisted of diabetic rats that were fed on normal diet.
     
    ♦ DIAB+MET (Diabetic plus 100 mg/kg metformin)
     
    In this group, rats were diabetic and were kept on 100 mg antidiabetic metformin drug.
     
    ♦ DIAB+25Q (Diabetic plus 25 mg Quercetin)
     
    Here, the rats were also diabetic and were provided with 25 mg Quercetin per kg per day.
     
    ♦ DIAB+50Q (Diabetic plus 50 mg Quercetin)
     
    This group comprised of diabetic rats, kept on 50 mg Quercetin.
     
    ♦NONDIAB+MET (Non-Diabetic plus 100 mg metformin)
     
    This experimental group included rats that were non- diabetic and were provided with 100 mg metformin drug.
     
    ♦ NONDIAB+25Q (Non-Diabetic plus 25 mg Quercetin)
     
    This group comprised of non-diabetic rats, kept on 25 mg Quercetin per kg per day.
     
    ♦ NONDIAB+50Q (Non-Diabetic plus 50 mg Quercetin)
     
    This non-diabetic group was given 50 mg Quercetin.
     
    Extraction of serum
     
    After 14 weeks trial period, blood was collected in vacutainer serum gel collection tubes. Following clotting of blood, these tubes were centrifuged at 2500 rpm for 10 minutes to collect serum, that was stored at -80oC for lipid profiling through micro lab-300 using Innoline commercial kits.

    mRNA extraction, cDNA synthesis, real time PCR
     
    The skeletal muscle, hepatic and adipose tissues were isolated from rats, washed with saline water and were stored at -80oC. Following the manufacturer’s instructions, Trizol reagent (Bioshop, Canada) was used for the extraction of RNA from the frozen samples of all the three tissues. The commercial RNA extraction kit (CO2203403, Ascend.bio, UK) was used. The extracted RNA was quantified using nanodrop (absorbance at 260 nm). RNA (1 µg) was changed into cDNA by the process of reverse transcription in a reaction volume of total 20 µl using cDNA kit (01317155, Thermo-scientific, Lithuania). The conventional  PCR was used for the conversion of RNA into cDNA, involving annealing at 25oC for 5 min, elongation step at 42oC for 60 min and enzyme inactivation at 70oC for 5 min. Quantitative PCR with the aid of SYBR Green master mix (Thermos-scientific) was operated on a Qiagen (Germany) following the instructions given by the manufacturer. The PCR was performed for a total of 45 cycles. The PCR product was monitored continuously by reading the fluorescence throughout the reaction. The gene expression results were analyzed on Rotor gene Q series software. Primer3 website was used for designing the primers. The GAPDH (housekeeping gene) was used for normalizing the mRNA levels. The delta Ct method was used for the relative quantification. The primers used in real time PCR are mentioned in Table 1.

    Table 1: Lipid metabolism genes and their primer sequences.


     
    Statistical analysis
     
    Results of the current study are represented as mean ± SEM. One way ANOVA was used for the analysis followed by Duncan multiple range test. Results were analyzed at level of significance (p<0.05), which was considered as significant for the particular study.

    RESULTS AND DISCUSSION

    Diabetes is regarded as one of the major health concerns worldwide. More than 90% of diabetes cases are attributed to type 2 diabetes mellitus and this particular disease is often ascribed as multifactorial origin chronic metabolic disorder. Diabetes is responsible for disturbing the lipid metabolism and causing inflammation in body. Polyphenols are phytochemicals that are present in vegetables and fruits, moreover they are further categorized into flavonoids, phenolic acids, stilbenes and lignans. Foods rich in Polyphenol and dietary supplements both are linked with a lower occurrence of lipid dependent cardiovascular disease (Costa et al., 2022). A common flavonoid, Quercetin, readily available in vegetables and fruits, is a potent antioxidant. Quercetin exhibit varied beneficial features, namely anti-diabetic, anti-dyslipidemia, anti-inflammatory, anti-cancer and antioxidant properties (Wang et al., 2022). Metformin hydrochloride with its anti-hyperglycemic and anti-dyslipidemia ability is also being in practice for diabetes treatment (Pournaghi et al., 2012; Szymczak-Pajor et al., 2022). Quercetin and Metformin play a crucial role in lipid metabolism through lipid metabolism gene expression. In our study, lipid profiling was done at the end of trial. Gene expression data was also recorded in liver, adipose tissue and skeletal muscles individually at the end of 14 weeks trial after slaughtering the rats.
     
    Ameliorative effect of quercetin and metformin on lipid profile in wistar rats
     
    In blood serum, lipid levels are good indicators of cardiovascular health. It is a well-documented fact that untreated type 2 diabetes mellitus results in elevated levels of LDL, triglycerides, VLDL and also decreased level of HDL, promoting progression to coronary artery diseases. Moreover, the deficiency of insulin in diabetes leads to a variety of irrationalities in regulatory and metabolic pathways, which ultimately results in accumulation of lipids namely TC and triglycerides in diabetic individuals. At the end of 14-week trial, serum was isolated from all the 64 albino rats for biochemical analysis. The results were revealed for serum TC, HDL, TG, LDL-C and VLDL-C (Table 2).

    Table 2: Lipid profile of Wistar rats in different treatment groups.


           
    In our research study, streptozotocin induced diabetic group displayed a considerable increase in LDL, VLDL, triglyceride, total cholesterol while a significant decrease in HDL levels as compared to CONTROL group rats. Following 14 week treatment, both Quercetin treated and Metformin treated diabetic rat groups showed a substantial reduction in LDL, VLDL, triglyceride, total cholesterol and elevated levels of HDL (Table 2). Earlier researches have also found improvement in lipid profile following Quercetin and Metformin treatment (Albasher et al., 2020; Ali et al., 2020; Abdelkader et al., 2020; Lai et al., 2021; Hacioglu et al., 2021; Chellian et al., 2022; Hu et al., 2022; Jiang et al., 2022; Rahmani et al., 2023).

    Gene expression of FAS, SREBP-1c and PPAR-α in liver
     
    In liver, FAS gene expression showed a significant difference (p<0.05) between CONTROL (1.03±0.09) and DIABETIC group (1.55±0.03), While other groups i.e. DIAB+MET (1.26±0.03), DIAB+25Q (1.31±0.02) and DIAB+50Q (1.19±0.04) showed a considerable difference (p<0.05) compared to DIABETIC group. In term of significance, all the three treatment groups (DIAB+MET, DIAB+25Q and DIAB+50Q) displayed statistically similar downregulation of FAS gene. There was a significant difference (p<0.05) between CONTROL and treatment groups (DIAB+MET, DIAB+25Q and DIAB+50Q). A non-significant difference was observed between CONTROL (1.03±0.09) and non-diabetic treatment groups; NONDIAB +MET (1.01±0.03), NONDIAB+25Q (1.03±0.05) and NONDIAB+50Q (0.98±0.04) (Fig 1).

    Fig 1: Gene expression of FAS, SREBP-1c and PPARa in liver.


           
    SREBP1-c gene expression in liver, also displayed a substantial difference (p<0.05) between CONTROL (1.05±0.12) and DIABETIC group (2.33±0.03), Whereas the treatment groups with diabetes i.e. DIAB+MET (1.31±0.05), DIAB+25Q (1.70±0.02) and DIAB+50Q (1.38±0.04) showed a considerable difference (p<0.05) compared to DIABETIC group. The DIAB+MET and DIAB+50Q groups exhibited major reduction in expression of SREBP1-c gene compared to DIAB+25Q. There was a significant difference (p<0.05) between CONTROL and treatment groups (DIAB+MET, DIAB+25Q and DIAB+50Q). A non-significant difference was observed between CONTROL (1.05±0.12) and non-diabetic treatment groups; NONDIAB+MET (0.96±0.02), NONDIAB+25Q (1.01±0.04) and NONDIAB+50Q  (0.98±0.04) (Fig 1).
           
    PPAR-α gene expression in liver, revealed a significant difference (p<0.05) between CONTROL (1.00±0.02) and DIABETIC group (0.54±0.04). The diabetic treatment groups i.e. DIAB+MET (0.87±0.06), DIAB+25Q (0.65±0.03) and DIAB+50Q (0.76±0.04) showed a considerable difference (p<0.05) compared to DIABETIC group, showing an improvement in liver PPAR-α gene expression compared to DIABETIC. The DIAB+MET group showed relatively better expression of PPAR-α gene compared to the other two diabetic treatment groups. A non-significant difference was observed between CONTROL (1.00±0.02) and non-diabetic treatment groups; NONDIAB+MET (1.00±0.04), NONDIAB+25Q (1.00±0.03) and NONDIAB+50Q (1.02±0.03) (Fig 1).
     
    Gene expression of FAS and PPAR-γ in adipose tissue
     
    In adipose tissue, FAS gene expression showed a significant variation (p<0.05) between CONTROL (1.01±0.06) and DIABETIC group (2.13±0.05), While other groups i.e. DIAB+MET (1.40±0.04), DIAB+25Q (1.72±0.04) and DIAB+50Q (1.28±0.03) showed a significant difference (p<0.05) compared to DIABETIC group. The most considerable downregulation of FAS gene was observed in DIAB+50Q group. There was a significant difference (p<0.05) between CONTROL and treatment groups (DIAB+MET, DIAB+25Q and DIAB+50Q). A non-significant difference was observed between CONTROL (1.01±0.06) and non-diabetic treatment groups; NONDIAB+MET (0.99±0.02), NONDIAB+25Q (1.01±0.03) and NONDIAB+50Q (0.97±0.03) (Fig 2).

    Fig 2: Gene expression of FAS and PPAR g in Adipose tissue.


           
    PPAR-γ gene expression in adipose, displayed a substantial difference (p<0.05) between CONTROL (1.11±0.18) and DIABETIC group (0.33±0.05), Whereas the treatment groups with diabetes i.e. DIAB+MET (0.84±0.05), DIAB+25Q (0.61±0.03) and DIAB+50Q (0.88±0.06) showed a significant difference (p<0.05) compared to DIABETIC group. A non-significant difference was observed between CONTROL (1.11±0.18) and non-diabetic treatment groups; NONDIAB+MET (1.15±0.04), NONDIAB+25Q (1.12±0.05) and NONDIAB+50Q (1.17±0.06) (Fig 2).
     
    Gene expression of PPAR-α in skeletal muscles
     
    PPAR-α gene expression in skeletal muscle, showed a significant variance (p<0.05) between CONTROL (1.00±0.03) and DIABETIC group (0.39±0.02). The diabetic treatment groups i.e. DIAB+MET (0.76±0.02), DIAB+25Q (0.59±0.03) and DIAB+50Q (0.80±0.03) showed a substantial difference (p<0.05) compared to DIABETIC group, indicating an improvement in skeletal muscle PPAR-α gene expression compared to DIABETIC. The DIAB+MET and DIAB+50Q groups showed similar upregulation of PPAR-α gene, with no significant difference between the two groups. A non-significant difference was observed between CONTROL (1.00±0.03) and non-diabetic treatment groups; NONDIAB +MET (1.01±0.02), NONDIAB+25Q (0.98±0.03) and NONDIAB+50Q (1.03±0.03) (Fig 3).

    Fig 3: Gene expression of PPAR-a in skeletal muscles.


           
    The expression data of PPAR-α in our study is similar with previously reported studies (Karimi-Sales et al., 2019; Malinska et al., 2019; Ting et al., 2018). PPAR-α is disseminated in several body tissues but is mainly present in liver, adipose and skeletal tissues. PPAR-α is involved in gene regulation specifically genes concerned with lipoprotein metabolism, lipid transport and fatty acid oxidation (Małodobra-Mazur et al., 2024).
           
    In term of PPAR-γ expression, the findings for Quercetin (Sun et al., 2015) are in line with previous studies. The experimental outcome of our study for PPAR-γ are also in accordance with previous study reporting an upregulation mediated by flavonoid  (Singh et al., 2018). Matsukawa et al., 2015, also found a significant upregulation of PPAR-γ in diabetic mice after treatment of mice with flavonoid, which is in accordance with our study results. PPAR-γ is regarded as the adipogenesis master regulator, as it increases the storage of lipid. PPAR-δ is also responsible for regulation of the change from glycolytic state to oxidative muscle fibres (Montaigne et al., 2021).
           
    SREBPs role in lipid metabolism is multifunctional and thereby numerous metabolic issues are related to dysregulation of SREBP for example dyslipidemia, type 2 diabetes, atherosclerosis and fatty liver disease (Li et al., 2023). SREBP-1c is a vital protein responsible for synthesizing fat and when SREBP-1c is activated then it further enhances adipogenic genes transcription such as fatty acid synthase and acetyl-CoA carboxylase, thus accelerating hepatic fat production (Gan et al., 2024). In case of gene expression involving SREBP-1c, Lee and Kim (2022) also found results relevant with out findings, reporting a down regulation of SREBP-1c in Quercetin treated groups in high fat diet model. Pengnet et al. (2022), also presented a relatable result, showing a downregulation of SREBP-1c in rats after flavonoid treatment. Karimi-Sales et al. (2019), presented a similar result, a downregulation of SREBP-1c in wistar rats following flavonoid treatment which is a similar trend as in our study.
           
    Our study outcome for FAS expression is in accordance with previously conducted studies (Afarin et al., 2024; Lee and Kim 2022). Fatty acid is synthesized by FAS in liver (Sahin et al., 2021) and other tissues namely adipose and skeletal muscle. In diabetes, there is an evident rise in FAS expression and thus consequently leading to dyslipidemia, as FAS is involved in lipid synthesis (Fang et al., 2024). In our study, diabetes induction also causes upregulation of FAS at a considerable level thereby affecting lipid metabolism while treatment with Quercetin and Metformin downregulated the FAS gene.

    CONCLUSION

    Our study outcome clearly showed that Quercetin flavonoid is equally effective in ameliorating lipid profile and its related gene expression as Metformin which is a synthetic drug. Moreover, the higher dose of 50 mg Quercetin showed comparatively better results in term of lipid profile and gene expression as compared to 25 mg dose.

    CONFLICT OF INTEREST

    All authors declared that there is no conflict of interest.

    REFERENCES


    1. Abdelkader, N.F., Eitah, H.E., Maklad, Y.A., Gamaleldin, A.A., Badawi, M.A. and Kenawy, S.A. (2020). New combination therapy of gliclazide and quercetin for protection against STZ- induced diabetic rats. Life Sciences. 247: 117458.

    2. Adnan, M. and Aasim, M. (2020). Prevalence of type 2 diabetes mellitus in adult population of Pakistan: A meta-analysis of prospective cross-sectional surveys. Annals of Global Health. 86(1).

    3. Afarin, R., Hatami, M., Monjezi, S., Bineshfar, F. and Ahangarpour, A. (2024). Suppression of TGF-β/Smad3 signaling pathway by Capparis spinosa and quercetin in a rat model of nonalcoholic steatohepatitis. Iranian Journal of Basic Medical Sciences. 27(9): 1096.

    4. Albasher, G., Alwahaibi, M., Abdel-Daim, M.M., Alkahtani, S. and Almeer, R. (2020). Protective effects of Artemisia judaica extract compared to metformin against hepatorenal injury in high-fat diet/streptozotocine-induced diabetic rats. Environmental Science and Pollution Research. 27: 40525-40536.

    5. Ali, A.M., Gabbar, M.A., Abdel-Twab, S.M., Fahmy, E.M., Ebaid, H., Alhazza, I.M. and Ahmed, O.M. (2020). Antidiabetic potency, antioxidant effects and mode of actions of Citrus reticulata fruit Peel hydroethanolic extract, hesperidin and quercetin in nicotinamide/streptozotocin induced wistar diabetic rats. Oxidative Medicine and Cellular Longevity. 2020(1): 1730492.

    6. Arvanitis, M. and Lowenstein, C.J. (2023). Dyslipidemia. Annals of Internal Medicine. 176(6): ITC81-ITC96.

    7. Bukhsh, A., Khan, T.M., Sarfraz Nawaz, M., Sajjad Ahmed, H., Chan, K.G. and Goh, B.H. (2019). Association of diabetes knowledge with glycemic control and self-care practices among Pakistani people with type 2 diabetes mellitus. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy. pp: 1409-1417.

    8. Chellian, J., Mak, K.K., Chellappan, D.K., Krishnappa, P. and Pichika, M.R. (2022). Quercetin and metformin synergistically reverse endothelial dysfunction in the isolated aorta of streptozotocin-nicotinamide-induceddiabetic rats. Scientific Reports. 12(1): 21393.

    9. Costa, P.C.T.D., de Souza, E.L., Lacerda, D.C., Cruz Neto, J.P.R., Sales, L.C.S.D., Silva Luis, C.C. and de Brito Alves, J.L. (2022). Evidence for quercetin as a dietary supplement for the treatment of cardio-metabolic diseases in pregnancy: A review in rodent models. Foods. 11(18): 2772.

    10. Devi, G., Dimri, U., Ajith, Y., Deepa, P.M., Yatoo, M.I., Gopalakrishnan, A. and Madhesh, E. (2023). The anti-oxidant and the anti-diabetic effects of terminalia chebula and withania somnifera in subclinically diabetic dogs. Indian Journal of Animal Research. 57(8): 1042-1050. doi: 10.18805/ IJAR.B-4355.

    11. Fang, Z., Liu, R., Xie, J. and He, J.C. (2024). Molecular mechanism of renal lipid accumulation in diabetic kidney disease. Journal of Cellular and Molecular Medicine. 28(11): e18364.

    12. Gan, L., Guo, H., Yang, Q., Zhou, X., Xie, Y., Ma, X. and Zuo, Z. (2024). Alkaline mineral complex water attenuates transportation-induced hepatic lipid metabolism dysregulation by AMPKα-SREBP-1c/PPARα pathways. International Journal of Molecular Sciences. 25(21): 11373.

    13. Guo, H., Wu, H. and Li, Z. (2023). The pathogenesis of diabetes. International Journal of Molecular Sciences. 24(8): 6978.

    14. Hacioglu, C., Kar, F., Kara, Y., Yucel, E., Donmez, D.B., Sentürk, H. and Kanbak, G. (2021). Comparative effects of metformin and Cistus laurifolius L. extract in streptozotocin-induced diabetic rat model: Oxidative, inflammatory, apoptotic and histopathological analyzes. Environmental Science and Pollution Research. 28(41): 57888-57901.

    15. Hosseini, A., Razavi, B.M., Banach, M. and Hosseinzadeh, H. (2021). Quercetin and metabolic syndrome: A review. Phytotherapy  Research. 35(10): 5352-5364.

    16. Hu, N., Zhang, Q., Wang, H., Yang, X., Jiang, Y., Chen, R. and Wang,  L. (2022). Comparative evaluation of the effect of metformin and insulin on gut microbiota and metabolome profiles of type 2 diabetic rats induced by the combination of streptozotocin and high-fat diet. Frontiers in Pharmacology. 12: 794103.

    17. Jiang, J.J., Zhang, G.F., Zheng, J.Y., Sun, J.H. and Ding, S.B. (2022). Targeting mitochondrial ROS-mediated ferroptosis by quercetin alleviates high-fat diet-induced hepatic lipotoxicity. Frontiers in Pharmacology. 13: 876550.

    18. Kane, J.P., Pullinger, C.R., Goldfine, I.D. and Malloy, M.J. (2021). Dyslipidemia and diabetes mellitus: Role of lipoprotein species and interrelated pathways of lipid metabolism in diabetes mellitus. Current Opinion in Pharmacology. 61: 21-27.

    19. Karimi-Sales, E., Ebrahimi-Kalan, A. and Alipour, M.R. (2019). Preventive effect of trans-chalcone on non-alcoholic steatohepatitis: Improvement of hepatic lipid metabolism. Biomedicine and Pharmacotherapy. 109: 1306-1312.

    20. Khajuria, P., Raghuwanshi, P., Rastogi, A., Koul, A.L., Zargar, R.  and Kour, S. (2018). Hepatoprotective effect of Seabuckthorn leaf extract in streptozotocin induced diabetes mellitus in Wistar rats. Indian Journal of Animal Research. 52(12): 1745-1750. doi: 10.18805/ijar.B-3439.

    21. Lai, L.L., Lu, H.Q., Li, W.N., Huang, H.P., Zhou, H.Y., Leng, E.N. and Zhang, Y.Y. (2021). Protective effects of quercetin and crocin in the kidneys and liver of obese sprague-dawley rats with Type 2 diabetes: Effects of quercetin and crocin on T2DM rats. Human and Experimental Toxicology. 40(4): 661-672.

    22. Lee, M.S. and Kim, Y. (2022). Effects of quercetin on the improvement of lipid metabolism through regulating hepatic AMPK and microRNA-21 in high cholesterol diet-fed mice. Journal of Nutrition and Health. 55(1): 36-46.

    23. Li, N., Li, X., Ding, Y., Liu, X., Diggle, K., Kisseleva, T. and Brenner, D.A. (2023). SREBP regulation of lipid metabolism in liver disease and therapeutic strategies. Biomedicines. 11(12): 3280.

    24. Malinska, H., Hüttl, M., Oliyarnyk, O., Markova, I., Poruba, M., Racova, Z. and Vecera, R. (2019). Beneficial effects of troxerutin on metabolic disorders in non-obese model of metabolic syndrome. Plos One. 14(8): e0220377.

    25. Małodobra-Mazur, M., O³dakowska, M. and Dobosz, T. (2024). Exploring PPAR gamma and PPAR alpha’s regulation role in metabolism via epigenetics mechanism. Biomolecules. 14(11): 1445.

    26. Manikandan, R., Anand, A.V., Sampathkumar, P. and Manoharan, N. (2018). Protective effect of Psidium guajava leaf ethanolic extract against streptozotocin-induced diabetes and lipidosis in rats. Indian Journal of Animal Research. 52(8): 1198-1205. doi: 10.18805/ijar.B-3337.

    27. Matsukawa, T., Inaguma, T., Han, J., Villareal, M.O. and Isoda, H. (2015). Cyanidin-3-glucoside derived from black soybeans ameliorate type 2 diabetes through the induction of differentiation of preadipocytes into smaller and insulin- sensitive adipocytes. The Journal of Nutritional Biochemistry. 26(8): 860-867.

    28. Montaigne, D., Butruille, L. and Staels, B. (2021). PPAR control of metabolism and cardiovascular functions. Nature Reviews Cardiology. 18(12): 809-823.

    29. Naseem, M., Ouguerram, K., Nazih, H., Rabbani, I., Zaneb, H., Rehman, H.U. and Tahir, S.K. (2020). Synergistic effect of extracts of Ginkgo biloba leaf and Panax ginseng root on carbohydrate and lipid metabolism gene expression in alloxan induced diabetic rats. JAPS: Journal of Animal and Plant Sciences. 30(6).

    30. Olimjonovna, K.O. (2024). Understanding the causes and risk factors of diabetes. Biologiya Va Kimyo Fanlari Ilmiy Jurnali. 2(5): 1-7.

    31. Pengnet, S., Sumarithum, P., Phongnu, N., Prommaouan, S., Kantip, N., Phoungpetchara, I. and Malakul, W. (2022). Naringin attenuates fructose-induced NAFLD progression in rats through reducing endogenous triglyceride synthesis and activating the Nrf2/HO-1 pathway. Frontiers in Pharma- cology. 13: 1049818.

    32. Pournaghi, P., Sadrkhanlou, R.A., Hasanzadeh, S. and Foroughi, A. (2012). An investigation on body weights, blood glucose levels and pituitary-gonadal axis hormones in diabetic and metformin-treated diabetic female rats. In Veterinary Research Forum. 3(2): 79. 

    33. Rahmani, A.H., Alsahli, M.A., Khan, A.A. and Almatroodi, S.A. (2023). Quercetin, a plant flavonol attenuates diabetic complications, renal tissue damage, renal oxidative stress and inflammation in streptozotocin-induced diabetic rats. Metabolites. 13(1): 130.

    34. Riddle, M.C., Cefalu, W.T., Evans, P.H., Gerstein, H.C., Nauck, M.A., Oh, W.K. and Twenefour, D. (2022). Consensus report: definition and interpretation of remission in type 2 diabetes. The Journal of Clinical Endocrinology and Metabolism. 107(1): 1-9.

    35. Sahin, K., Orhan, C., Kucuk, O., Erten, F., Tuzcu, M., Sahin, N. and Komorowski, J.R. (2021). Effects of magnesium biotinate supplementation on serum insulin, glucose and lipid parameters along with liver protein levels of lipid metabolism in rats. Magnesium Research. 34(1): 9-19.

    36. Singh, A.K., Raj, V., Keshari, A.K., Rai, A., Kumar, P., Rawat, A. and Saha, S. (2018). Isolated mangiferin and naringenin exert antidiabetic effect via PPARγ/GLUT4 dual agonistic action with strong metabolic regulation. Chemico-Biological Interactions. 280: 33-44.

    37. Sun, L., Li, E., Wang, F., Wang, T., Qin, Z., Niu, S. and Qiu, C. (2015). Quercetin increases macrophage cholesterol efflux to inhibit foam cell formation through activating PPARγ- ABCA1 pathway. International Journal of Clinical and Experimental Pathology. 8(9): 10854.

    38. Szymczak-Pajor, I., Wenclewska, S. and Śliwińska, A. (2022). Metabolic action of metformin. Pharmaceuticals. 15(7): 810.

    39. Ting, Y., Chang, W.T., Shiau, D.K., Chou, P.H., Wu, M.F. and Hsu, C.L. (2018). Antiobesity efficacy of quercetin-rich supplement on diet-induced obese rats: Effects on body composition, serum lipid profile and gene expression. Journal of Agricultural and Food Chemistry. 66(1): 70-80.

    40. Umar, H.B. and Utari, D.M. (2019). Differences in consumption of flavonoid phytochemicals toward total cholesterol and LDL-cholesterol levels in dyslipidemia and non-dyslipidemia groups of minangkabau ethnic women. Indian Journal of Public Health Research and Development. 10(12).

    41. Wang, D., Li, Y., Zhai, Q.Q., Zhu, Y.F., Liu, B.Y. and Xu, Y. (2022). Quercetin ameliorates testosterone secretion disorder by inhibiting endoplasmic reticulum stress through the miR-1306-5p/HSD17B7 axis in diabetic rats. Bosnian Journal of Basic Medical Sciences. 22(2): 191. 

    42. Yan, L., Vaghari-Tabari, M., Malakoti, F., Moein, S., Qujeq, D., Yousefi, B. and Asemi, Z. (2023). Quercetin: An effective polyphenol in alleviating diabetes and diabetic complications. Critical Reviews in Food Science and Nutrition. 63(28): 9163- 9186. 
    In this Article
      Contact us
      Follow us
      Published In
      Indian Journal of Animal Research

      Editorial Board

      View all (0)