Quercetin mitigates hepatic injury by inhibiting TGF-β/p-Smad3 signaling pathway activation and enhancing AMPK activity in type 2 diabetic rats

Abstract

Objective: To investigate the effects of quercetin on inflammatory signaling pathways and hepatic oxidative injury using a streptozotocin-induced liver injury model.

Methods: Four groups of 32 rats were used in this study: three groups were given streptozotocin to induce diabetes, and one group was given a normal control. For treatment groups, each group received either metformin (200 mg/kg body weight) or quercetin (50 mg/kg body weight) for a month. The expression of SREBP1c was detected by quantitative RT-PCR and fibrosis-related proteins (TGF-β and p-Smad3) was evaluated by Western blot. Furthermore, MCP and IL-1β were determined by ELISA.

Results: Quercetin significantly reduced insulin and glucose levels. Besides, it reduced the serum levels of ALT, AST, and ALP, improved lipid profile, lowered the MDA level, increased SOD activity, decreased the rise in MCP-1 and IL-1β, inhibited the TGF-β/Smad3 signaling pathway, decreased α-SMA and SREBP1c expression, and increased AMPK (P<0.05).

Conclusions: Quercetin could significantly mitigate hepatic damage by modulating the expression of pro-inflammatory cytokines and fibrosis markers.

Keywords

Quercetin / Metformin / Streptozotocin / Liver injury / TGF-β/Smad signaling / AMPK / SREBPlc

Conflict of interest statement

The authors declare that there is no conflict of interest.

Acknowledgments

Sincerely the authors acknowledge Dr. Ahmed Osman (Professor of histopathology, Faculty of Veterinary Medicine, Cairo University) for accomplishing the histopathological examination in the present study.

Funding

The authors received no extramural funding for the study.

Data availability statement

The data supporting the findings of this study are available from the corresponding author upon request.

Authors’ contributions

HFBG, MKM, FSMM, ESAA, and OARAZ contributed to the design and implementation of the research, the analysis of the results, and the writing of the manuscript. All authors discussed the results and commented on the manuscript.

Publisher’s note

The Publisher of the Journal remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

[1] Forouhi NG, Wareham NJ. Epidemiology of diabetes. Medicine 2014; 42(12): 698-702.

[2] Lee CH, Lui DT, Lam KS. Non-alcoholic fatty liver disease and type 2 diabetes: An update. J Diabetes Investig 2022; 13(6): 930-940.

[3] Kim KS, Hong S, Han K, Park CY. Association of non-alcoholic fatty liver disease with cardiovascular disease and all cause death in patients with type 2 diabetes mellitus: Nationwide population based study. BMJ 2024; 384: e076388.

[4] Erario M, Younossi ZM. Nonalcoholic fatty liver disease in adults:The impact of diet and other therapeutic interventions on clinical and histological outcomes. In:Preedy V, Hunter LA, Patel V (eds.) Diet quality. New York, NY,USA: Nutrition and Health, Humana Press; 2013.

[5] Feng G, Valenti L, Wong VW, Fouad YM, Yilmaz Y, Kim W, et al. Recompensation in cirrhosis: Unravelling the evolving natural history of nonalcoholic fatty liver disease. Nat Rev Gastroenterol Hepatol 2024; 21(1): 46-56.

[6] Machado M, Cortez-Pinto H. Non-alcoholic fatty liver disease: What the clinician needs to know. World J Gastroenterol 2014; 20(36): 12956-12980.

[7] Elpek GÖ. Cellular and molecular mechanisms in the pathogenesis of liver fibrosis: An update. World J Gastroenterol 2014; 20(23): 7260-7276.

[8] Schwenger KJ, Allard JP. Clinical approaches to non-alcoholic fatty liver disease. World J Gastroenterol 2014; 20(7): 1712-1723.

[9] Van Wagner LB, Rinella ME. The role of insulin-sensitizing agents in the treatment of nonalcoholic steatohepatitis. Therap Adv Gastroenterol 2011; 4(4): 249-263.

[10] Zhang X, Ruan J, He Y, Xu A, Fang Y, Zhang Q, et al. Dietary inflammatory index and the risks of non-alcoholic fatty liver disease: A systematic review and meta-analysis. Front Nutr 2024; 11: 1388557.

[11] Sodum N, Kumar G, Bojja SL, Kumar N, Rao CM. Epigenetics in NAFLD/NASH: Targets and therapy. Pharmacol Res 2021; 167: 105484.

[12] Rahmani AH, Alsahli MA, Khan AA, Almatroodi SA. Quercetin, a plant flavonol attenuates diabetic complications, renal tissue damage, renal oxidative stress and inflammation in streptozotocin-induced diabetic rats. Metabolites 2023; 13(1). doi: 10.3390/metabo13010130.

[13] Wu Q, Chen Z, Ding Y, Tang Y, Cheng Y. Protective effect of traditional Chinese medicine on non-alcoholic fatty liver disease and liver cancer by targeting ferroptosis. Front Nutr 2022; 9: 1033129.

[14] Dhasarathan P, Theriappan P. Evaluation of anti-diabetic activity of Strychonous potatorum in alloxan induced diabetic rats. J Med Sci 2011; 2: 670-674.

[15] Lin Z, Cheng X, Zheng H. Umbelliferon: A review of its pharmacology, toxicity and pharmacokinetics. Inflammopharmacology 2023; 31(4): 1731-1750.

[16] Dabeek WM, Marra MV. Dietary quercetin and kaempferol: Bioavailability and potential cardiovascular-related bioactivity in humans. Nutrients 2019; 11: 2288.

[17] Saeedi-Boroujeni A, Mahmoudian-Sani MR. Anti-inflammatory potential of quercetin in COVID-19 treatment. J Inflamm 2021; 18(1): 3.

[18] Alzohairy MA, Khan AA, Ansari MA, Babiker AY, Alsahli MA, Almatroodi SA, et al. Protective effect of quercetin, a flavonol against benzo(a)pyrene-induced lung injury via inflammation, oxidative stress, angiogenesis and cyclooxygenase-2 signalling molecule. Appl Sci 2021; 11: 8675.

[19] Qiang G, Yang X, Xuan Q, Shi L, Zhang H, Chen B, et al. Salvianolic acid a prevents the pathological progression of hepatic fibrosis in high-fat diet-fed and streptozotocin-induced diabetic rats. Am J Chin Med 2014; 42(5): 1183-1198.

[20] Ola MS, Ahmed MM, Shams S, Al-Rejaie SS. Neuroprotective effects of quercetin in diabetic rat retina. Saudi J Biol Sci 2017; 24(6): 1186-1194.

[21] Al Za’abi M, Ali BH, Al Suleimani Y, Adham SA, Ali H, Manoj P, et al. The effect of metformin in diabetic and non-diabetic rats with experimentally-induced chronic kidney disease. Biomolecules 2021; 11(6): 814.

[22] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 2001; 25(4): 402-408.

[23] Bancroft JD, Stevens A, Turner DR. Theory and practice of histological techniques. 4th Ed. London: Churchill Livingston; 2013.

[24] Plaa G, Charbonneau M. Detection and evaluation of chemically induced liver injury. In: Hayes AW (ed.) Principles and methods of toxicology. New York: Raven Press; 1994; p. 841-846.

[25] Park HJ, Kim HG, Wang JH, Choi MK, Han JM, Lee JS, et al. Comparison of TGF-β PDGF, and CTGF in hepatic fibrosis models using DMN, Ccl₄, and TAA. Drug Chem Toxicol 2016; 39(1): 111-118.

[26] En Li, Cho E, Ang CZ, Quek J, Fu CE, Lim LKE, et al. Global prevalence of non-alcoholic fatty liver disease in type 2 diabetes mellitus: An updated systematic review and meta-analysis. Gut 2023; 72(11): 2138-2148.

[27] Mohamed J, Nazratun Nafizah AH, Zariyantey AH, Budin SB. Mechanisms of diabetes-induced liver damage: The role of oxidative stress and inflammation. Sultan Qaboos Univ Med J 2016; 16(2): e132-e141.

[28] Abdelbassir AF, Abdelmageed AD, Ali HA, Ali HA, Alharthy H. Biochemical and molecular effects of the Laurel plant extract and olive leaves extract in diabetic rats induced by streptozotocin. Benha Vet Med J 2024; 47: 33-39.

[29] Rateb MO, Abdelmagid AD, Abdelhamid OM, Said AM. Evaluation of the antidiabetic and hepatoprotective effect of Origanum majorana leaf extract and its nanoparticles in streptozotocin-induced diabetes in rats. Benha Vet Med J 2024; 46: 23-27.

[30] Day CP, James OF. Steatohepatitis: A tale of two “hits”? Gastroenterology 1998; 114(4): 842-845.

[31] Zhang XQ, Zhao D, Ma YD, Wang YC, Zhang LX, Guo WJ, et al. Impact of disturbed glucose homeostasis regulated by AMPK in endometrium on embryo implantation in diabetes mice. Reprod Sci 2020; 27(9): 1752-1757.

[32] Behrouz V, Dastkhosh A, Hedayati M, Sedaghat M, Sharafkhah M, Sohrab G. The effect of crocin supplementation on glycemic control, insulin resistance and active AMPK levels in patients with type 2 diabetes: a pilot study. Diabetol Metab Syndr 2020; 12: 59.

[33] Watanabe S, Li YS, Kawasaki Y, Ootsuyama Y, Kawai K. Health examination results and work environment factors affecting urinary 8-hydroxy-2’-deoxyguanosine levels. J Occup Health 2021; 63(1): e12210.

[34] Oryan A, Bemani E, Bahrami S. Anti-leishmanial, immunomodulatory and anti-oxidative activity of quercetin against cutaneous leishmaniasis caused by Leishmania major. Asian Pac J Trop Biomed 2023; 13(1): 26-34.

[35] Chenna H, Khelef Y, Halimi I, Yilmaz MA, Çakir O, Djouder C, et al. Potential hepatoprotective effect of matricaria pubescens on high-fat diet- induced non-alcoholic fatty liver disease in rats. Chem Biodivers 2024; 21(4): e202302005.

[36] Tateya S, Kim F, Tamori Y. Recent advances in obesity-induced inflammation and insulin resistance. Front Endocrinol (Lausanne) 2013; 4: 93.

[37] Xiao C, Li J, Dong X, He X, Niu X, Liu C, et al. Anti-oxidative and TNF-α suppressive activities of puerarin derivative (4AC) in RAW264.7 cells and collagen-induced arthritic rats. Eur J Pharmacol 2011; 666(1- 3): 242-250.

[38] Hernandez-Gea V, Friedman SL. Pathogenesis of liver fibrosis. Annu Rev Pathol 2011; 6: 425-456.

[39] Qian K, Lei X, Liu G, Fang Y, Xie C, Wu X, et al. Transient receptor potential vanilloid-1 (TRPV1) alleviates hepatic fibrosis via TGF-β signaling. Dis Markers 2022; 2022: 3100943.

[40] Zheng W, Song J, Zhang Y, Chen S, Ruan H, Fan C. Metformin prevents peritendinous fibrosis by inhibiting transforming growth factor-β signaling. Oncotarget 2017; 8(60): 101784-101794.