Beneficial Effect of Metformin, Quercetin, and Resveratrol Combination on High Glucose-Induced lipogenesis in HepG2 Cells
Abstract
Objectives: It has been reported that Metformin (MET), Resveratrol (RSV), and Quercetin (QRS) possess anti-lipogenic effects. This study aimed to investigate the combined effects of these compounds on lipid accumulation in HepG2 cells treated with high glucose (HG).
Methods: HepG2 cells were treated with HG (33 mM), and different concentrations of MET, QRS, and RSV. The cytotoxic effects of these compounds were determined by an MTT assay. Changes in total lipid content and triglyceride (TG) levels were measured using Oil Red O staining and a triglyceride assay kit, respectively. The expression of fatty acid synthetase (FAS) and sterol regulatory element-binding protein 1c (SREBP-1c) was evaluated by quantitative real-time PCR.
Results: MET at doses 1, 2, and 5 mM, QRS at doses 5, 10, and 20 µM, and RSV at doses 25 and 50 µM could decrease total lipid content and triglyceride levels in HepG2 cells. The EC50 (half maximal effective concentration) from Oil red O staining results were MET: 1.786 mM, QRS: 8.132 μM, and RSV: 10.9 μM. The combination of MET (mM), QRS (µM), and RSV (µM) at ratios of (2:20:50), (1:10:25), and (0.5:5:10), could reduce lipogenesis greater than that observed with each of the individual compounds of MET, QRS, or RSV or the double combinations of MET+QRS or MET+RSV. In addition, combined treatment of MET (0.5mM), QRS (5 µM), and RSV (10 µM) was able to decrease SREBP-1c and FAS genes expression in HG-treated cells.
Conclusion: The combination of MET, QRS, and RSV could inhibit lipid accumulation in HepG2 cells by reducing total lipid content, triglyceride levels, and the expression of the genes involved in lipogenesis.
1. Lin, X., Y. Xu, X. Pan, J. Xu, Y. Ding, X. Sun, et al., Global, regional, and national burden and trend of diabetes in 195 countries and territories: an analysis from 1990 to 2025. Scientific reports, 2020. 10(1): p. 14790.
2. Meshkani, R., M. Taghikhani, B. Larijani, S. Khatami, E. Khoshbin, and K. Adeli, The relationship between homeostasis model assessment and cardiovascular risk factors in Iranian subjects with normal fasting glucose and normal glucose tolerance. Clinica chimica acta, 2006. 371(1-2): p. 169-175.
3. Hamzeh Saberi , G.T., Reza Meshkani The ENPP1 K121Q polymorphism is associated with obesity-related parameters in Iranian normoglycemic male subjects. Acta Biochimica Iranica, 2023. 1(1): p. 20-25.
4. Meshkani, R. and K. Adeli, Hepatic insulin resistance, metabolic syndrome and cardiovascular disease. Clinical biochemistry, 2009. 42(13-14): p. 1331-1346.
5. Khodabandehloo, H., S. Gorgani-Firuzjaee, G. Panahi, and R. Meshkani, Molecular and cellular mechanisms linking inflammation to insulin resistance and β-cell dysfunction. Translational Research, 2016. 167(1): p. 228-256.
6. Lin, M., D. Gordon, and J.R. Wetterau, Microsomal triglyceride transfer protein (MTP) regulation in HepG2 cells: insulin negatively regulates MTP gene expression. Journal of lipid research, 1995. 36(5): p. 1073-1081.
7. Reaven, G. and C. Mondon, Effect of in vivo plasma insulin levels on the relationship between perfusate free fatty acid concentration and triglyceride secretion by perfused rat livers. Hormone and metabolic research, 1984. 16(05): p. 230-232.
8. Garber, A.J., T.G. Duncan, A.M. Goodman, D.J. Mills, and J.L. Rohlf, Efficacy of metformin in type II diabetes: results of a double-blind, placebo-controlled, dose-response trial. The American journal of medicine, 1997. 103(6): p. 491-497.
9. Zare, M., G. Panahi, M. Koushki, Z. Mostafavi-Pour, and R. Meshkani, Metformin reduces lipid accumulation in HepG2 cells via downregulation of miR-33b. Archives of Physiology and Biochemistry, 2022. 128(2): p. 333-340.
10. Nima Taghizadeh , S.M., Vahid Saeedi, Ladan Haghighi, Mona Nourbakhsh, Mitra Nourbakhsh, Maryam Razzaghy Azar, Association between Steroid Hormones and Insulin Resistance in Patients with Polycystic Ovary Syndrome. Acta Biochimica Iranica, 2023. 1: p. 1.
11. Turner, R.C., C.A. Cull, V. Frighi, R.R. Holman, U.P.D.S. Group, and U.P.D.S. Group, Glycemic control with diet, sulfonylurea, metformin, or insulin in patients with type 2 diabetes mellitus: progressive requirement for multiple therapies (UKPDS 49). Jama, 1999. 281(21): p. 2005-2012.
12. Okayasu, S., K. Kitaichi, A. Hori, T. Suwa, Y. Horikawa, M. Yamamoto, et al., The evaluation of risk factors associated with adverse drug reactions by metformin in type 2 diabetes mellitus. Biological and Pharmaceutical Bulletin, 2012. 35(6): p. 933-937.
13. Niedzwiecki, A., M.W. Roomi, T. Kalinovsky, and M. Rath, Anticancer efficacy of polyphenols and their combinations. Nutrients, 2016. 8(9): p. 552.
14. Zamani‐Garmsiri, F., S. Emamgholipour, S. Rahmani Fard, G. Ghasempour, R. Jahangard Ahvazi, and R. Meshkani, Polyphenols: potential anti‐inflammatory agents for treatment of metabolic disorders. Phytotherapy Research, 2022. 36(1): p. 415-432.
15. Bischoff, S.C., Quercetin: potentials in the prevention and therapy of disease. Current Opinion in Clinical Nutrition & Metabolic Care, 2008. 11(6): p. 733-740.
16. Khodabandehloo, H., S. Seyyedebrahimi, E.N. Esfahani, F. Razi, and R. Meshkani, Resveratrol supplementation decreases blood glucose without changing the circulating CD14+ CD16+ monocytes and inflammatory cytokines in patients with type 2 diabetes: a randomized, double-blind, placebo-controlled study. Nutrition research, 2018. 54: p. 40-51.
17. Seyyedebrahimi, S., H. Khodabandehloo, E. Nasli Esfahani, and R. Meshkani, The effects of resveratrol on markers of oxidative stress in patients with type 2 diabetes: a randomized, double-blind, placebo-controlled clinical trial. Acta diabetologica, 2018. 55: p. 341-353.
18. Sattar Gorgani-Firuzjaee, R.M., Resveratrol reduces high glucose-induced de-novo lipogenesis through mTOR mediated induction of autophagy in HepG2 cells. Acta Biochimica Iranica, 2023. 1(1): p. 32-39.
19. Gorgani-Firuzjaee, S. and R. Meshkani, SH2 domain-containing inositol 5-phosphatase (SHIP2) inhibition ameliorates high glucose-induced de-novo lipogenesis and VLDL production through regulating AMPK/mTOR/SREBP1 pathway and ROS production in HepG2 cells. Free radical biology and medicine, 2015. 89: p. 679-689.
20. Roya Jahangard , S.S.S.E., Akram Vatannejad, Reza Meshkani Autophagy protects peripheral blood mononuclear cells from high glucose-induced inflammation and apoptosis. Acta Biochimica Iranica, 2023. 1(1): p. 40-49.
21. Cahn, A. and W.T. Cefalu, Clinical considerations for use of initial combination therapy in type 2 diabetes. Diabetes Care, 2016. 39(Supplement_2): p. S137-S145.
22. Bruckbauer, A. and M.B. Zemel, Synergistic effects of metformin, resveratrol, and hydroxymethylbutyrate on insulin sensitivity. Diabetes, metabolic syndrome and obesity: targets and therapy, 2013: p. 93-102.
23. Tehrani, S.S., G. Goodarzi, G. Panahi, F. Zamani-Garmsiri, and R. Meshkani, The combination of metformin with morin alleviates hepatic steatosis via modulating hepatic lipid metabolism, hepatic inflammation, brown adipose tissue thermogenesis, and white adipose tissue browning in high-fat diet-fed mice. Life Sciences, 2023. 323: p. 121706.
24. Dludla, P.V., S. Silvestri, P. Orlando, K.B. Gabuza, S.E. Mazibuko-Mbeje, T.M. Nyambuya, et al., Exploring the comparative efficacy of metformin and resveratrol in the management of diabetes-associated complications: a systematic review of preclinical studies. Nutrients, 2020. 12(3): p. 739.
25. Belew, G.D. and J.G. Jones, De novo lipogenesis in non‐alcoholic fatty liver disease: Quantification with stable isotope tracers. European journal of clinical investigation, 2022. 52(3): p. e13733.
26. Sanders, F.W. and J.L. Griffin, De novo lipogenesis in the liver in health and disease: more than just a shunting yard for glucose. Biological Reviews, 2016. 91(2): p. 452-468.
27. Knebel, B., P. Fahlbusch, M. Dille, N. Wahlers, S. Hartwig, S. Jacob, et al., Fatty liver due to increased de novo lipogenesis: alterations in the hepatic peroxisomal proteome. Front Cell Dev Biol. 2019; 7: 248. 2019.
28. Damiano, F., L. Giannotti, G.V. Gnoni, L. Siculella, and A. Gnoni, Quercetin inhibition of SREBPs and ChREBP expression results in reduced cholesterol and fatty acid synthesis in C6 glioma cells. The international journal of biochemistry & cell biology, 2019. 117: p. 105618.
29. Hosseini, H., M. Teimouri, M. Shabani, M. Koushki, R.B. Khorzoughi, F. Namvarjah, et al., Resveratrol alleviates non-alcoholic fatty liver disease through epigenetic modification of the Nrf2 signaling pathway. The international journal of biochemistry & cell biology, 2020. 119: p. 105667.
2. Meshkani, R., M. Taghikhani, B. Larijani, S. Khatami, E. Khoshbin, and K. Adeli, The relationship between homeostasis model assessment and cardiovascular risk factors in Iranian subjects with normal fasting glucose and normal glucose tolerance. Clinica chimica acta, 2006. 371(1-2): p. 169-175.
3. Hamzeh Saberi , G.T., Reza Meshkani The ENPP1 K121Q polymorphism is associated with obesity-related parameters in Iranian normoglycemic male subjects. Acta Biochimica Iranica, 2023. 1(1): p. 20-25.
4. Meshkani, R. and K. Adeli, Hepatic insulin resistance, metabolic syndrome and cardiovascular disease. Clinical biochemistry, 2009. 42(13-14): p. 1331-1346.
5. Khodabandehloo, H., S. Gorgani-Firuzjaee, G. Panahi, and R. Meshkani, Molecular and cellular mechanisms linking inflammation to insulin resistance and β-cell dysfunction. Translational Research, 2016. 167(1): p. 228-256.
6. Lin, M., D. Gordon, and J.R. Wetterau, Microsomal triglyceride transfer protein (MTP) regulation in HepG2 cells: insulin negatively regulates MTP gene expression. Journal of lipid research, 1995. 36(5): p. 1073-1081.
7. Reaven, G. and C. Mondon, Effect of in vivo plasma insulin levels on the relationship between perfusate free fatty acid concentration and triglyceride secretion by perfused rat livers. Hormone and metabolic research, 1984. 16(05): p. 230-232.
8. Garber, A.J., T.G. Duncan, A.M. Goodman, D.J. Mills, and J.L. Rohlf, Efficacy of metformin in type II diabetes: results of a double-blind, placebo-controlled, dose-response trial. The American journal of medicine, 1997. 103(6): p. 491-497.
9. Zare, M., G. Panahi, M. Koushki, Z. Mostafavi-Pour, and R. Meshkani, Metformin reduces lipid accumulation in HepG2 cells via downregulation of miR-33b. Archives of Physiology and Biochemistry, 2022. 128(2): p. 333-340.
10. Nima Taghizadeh , S.M., Vahid Saeedi, Ladan Haghighi, Mona Nourbakhsh, Mitra Nourbakhsh, Maryam Razzaghy Azar, Association between Steroid Hormones and Insulin Resistance in Patients with Polycystic Ovary Syndrome. Acta Biochimica Iranica, 2023. 1: p. 1.
11. Turner, R.C., C.A. Cull, V. Frighi, R.R. Holman, U.P.D.S. Group, and U.P.D.S. Group, Glycemic control with diet, sulfonylurea, metformin, or insulin in patients with type 2 diabetes mellitus: progressive requirement for multiple therapies (UKPDS 49). Jama, 1999. 281(21): p. 2005-2012.
12. Okayasu, S., K. Kitaichi, A. Hori, T. Suwa, Y. Horikawa, M. Yamamoto, et al., The evaluation of risk factors associated with adverse drug reactions by metformin in type 2 diabetes mellitus. Biological and Pharmaceutical Bulletin, 2012. 35(6): p. 933-937.
13. Niedzwiecki, A., M.W. Roomi, T. Kalinovsky, and M. Rath, Anticancer efficacy of polyphenols and their combinations. Nutrients, 2016. 8(9): p. 552.
14. Zamani‐Garmsiri, F., S. Emamgholipour, S. Rahmani Fard, G. Ghasempour, R. Jahangard Ahvazi, and R. Meshkani, Polyphenols: potential anti‐inflammatory agents for treatment of metabolic disorders. Phytotherapy Research, 2022. 36(1): p. 415-432.
15. Bischoff, S.C., Quercetin: potentials in the prevention and therapy of disease. Current Opinion in Clinical Nutrition & Metabolic Care, 2008. 11(6): p. 733-740.
16. Khodabandehloo, H., S. Seyyedebrahimi, E.N. Esfahani, F. Razi, and R. Meshkani, Resveratrol supplementation decreases blood glucose without changing the circulating CD14+ CD16+ monocytes and inflammatory cytokines in patients with type 2 diabetes: a randomized, double-blind, placebo-controlled study. Nutrition research, 2018. 54: p. 40-51.
17. Seyyedebrahimi, S., H. Khodabandehloo, E. Nasli Esfahani, and R. Meshkani, The effects of resveratrol on markers of oxidative stress in patients with type 2 diabetes: a randomized, double-blind, placebo-controlled clinical trial. Acta diabetologica, 2018. 55: p. 341-353.
18. Sattar Gorgani-Firuzjaee, R.M., Resveratrol reduces high glucose-induced de-novo lipogenesis through mTOR mediated induction of autophagy in HepG2 cells. Acta Biochimica Iranica, 2023. 1(1): p. 32-39.
19. Gorgani-Firuzjaee, S. and R. Meshkani, SH2 domain-containing inositol 5-phosphatase (SHIP2) inhibition ameliorates high glucose-induced de-novo lipogenesis and VLDL production through regulating AMPK/mTOR/SREBP1 pathway and ROS production in HepG2 cells. Free radical biology and medicine, 2015. 89: p. 679-689.
20. Roya Jahangard , S.S.S.E., Akram Vatannejad, Reza Meshkani Autophagy protects peripheral blood mononuclear cells from high glucose-induced inflammation and apoptosis. Acta Biochimica Iranica, 2023. 1(1): p. 40-49.
21. Cahn, A. and W.T. Cefalu, Clinical considerations for use of initial combination therapy in type 2 diabetes. Diabetes Care, 2016. 39(Supplement_2): p. S137-S145.
22. Bruckbauer, A. and M.B. Zemel, Synergistic effects of metformin, resveratrol, and hydroxymethylbutyrate on insulin sensitivity. Diabetes, metabolic syndrome and obesity: targets and therapy, 2013: p. 93-102.
23. Tehrani, S.S., G. Goodarzi, G. Panahi, F. Zamani-Garmsiri, and R. Meshkani, The combination of metformin with morin alleviates hepatic steatosis via modulating hepatic lipid metabolism, hepatic inflammation, brown adipose tissue thermogenesis, and white adipose tissue browning in high-fat diet-fed mice. Life Sciences, 2023. 323: p. 121706.
24. Dludla, P.V., S. Silvestri, P. Orlando, K.B. Gabuza, S.E. Mazibuko-Mbeje, T.M. Nyambuya, et al., Exploring the comparative efficacy of metformin and resveratrol in the management of diabetes-associated complications: a systematic review of preclinical studies. Nutrients, 2020. 12(3): p. 739.
25. Belew, G.D. and J.G. Jones, De novo lipogenesis in non‐alcoholic fatty liver disease: Quantification with stable isotope tracers. European journal of clinical investigation, 2022. 52(3): p. e13733.
26. Sanders, F.W. and J.L. Griffin, De novo lipogenesis in the liver in health and disease: more than just a shunting yard for glucose. Biological Reviews, 2016. 91(2): p. 452-468.
27. Knebel, B., P. Fahlbusch, M. Dille, N. Wahlers, S. Hartwig, S. Jacob, et al., Fatty liver due to increased de novo lipogenesis: alterations in the hepatic peroxisomal proteome. Front Cell Dev Biol. 2019; 7: 248. 2019.
28. Damiano, F., L. Giannotti, G.V. Gnoni, L. Siculella, and A. Gnoni, Quercetin inhibition of SREBPs and ChREBP expression results in reduced cholesterol and fatty acid synthesis in C6 glioma cells. The international journal of biochemistry & cell biology, 2019. 117: p. 105618.
29. Hosseini, H., M. Teimouri, M. Shabani, M. Koushki, R.B. Khorzoughi, F. Namvarjah, et al., Resveratrol alleviates non-alcoholic fatty liver disease through epigenetic modification of the Nrf2 signaling pathway. The international journal of biochemistry & cell biology, 2020. 119: p. 105667.
| Files | ||
| Issue | Vol 1 No 2 (2023) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/abi.v1i2.14107 | |
| Keywords | ||
| Diabetes Lipogenesis Metformin Quercetin Resveratrol HepG2 Combination Liver Non-alcoholic fatty liver disease | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Babaei Khorzoughi R, Meshkani R. Beneficial Effect of Metformin, Quercetin, and Resveratrol Combination on High Glucose-Induced lipogenesis in HepG2 Cells. ABI. 2023;1(2):96-104.
