Effects of simultaneous intervention of aerobic training and berberine supplementation on AMPK and GLUT4 gene expression in the quadriceps muscle of diabetic male rats

Zaefarani Arani, Mahboobe; Ramezani, Javad; Naghibi, Saeid

doi:10.48308/joeppa.2024.233187.1194

Effects of simultaneous intervention of aerobic training and berberine supplementation on AMPK and GLUT4 gene expression in the quadriceps muscle of diabetic male rats

Document Type : Original Article

Authors

Department of Sports Sciences, Faculty of Humanities, Payame Noor University, Tehran, Iran

10.48308/joeppa.2024.233187.1194

Abstract

Background and Purpose: Diabetes is a metabolic disease associated with increased blood glucose. Based on the results of previous research, physical activity, and herbal supplements containing antioxidants and anti-inflammatory agents play an important role in controlling blood glucose in diabetics. GLUT4 as a glucose transporter and AMPK as an insulin-independent biological pathway play an important role in energy homeostasis in the body. This study aimed to investigate the simultaneous intervention of aerobic training and berberine supplementation on GLUT4 and AMPK gene expression in the quadriceps muscle of diabetic male rats.

Materials and Methods: In this experimental research, 35 male Wistar rats (age 8 weeks, weight 242.15±10.32 gr) were randomly divided into five groups (n=7) including healthy control, diabetic control, diabetes + berberine, diabetes + aerobic training, and diabetes + aerobic training + berberine. Diabetes was induced by one-step intraperitoneal injection of STZ (60 mg/kg). Blood glucose above 300 mg/dL meant successful induction of diabetes. The training groups performed a treadmill protocol for six weeks, five days a week, with a gradual increase in speed and time. Berberine supplementation with a dose of 50 mg per kilogram of body weight was done by gavage on all days of week and in training days one hour before training. Forty-eight hours after the last training session, rats were deeply anesthetized by intraperitoneal injection of ketamine (75 mg/kg body weight) and xylazine (10 mg/kg body weight) and quadriceps muscle tissue samples were collected. RT-PCR technique was used to measure the relative expression of GLUT4 and AMPK genes. The data were analyzed using SPSS software. One-way analysis of variance and Tukey's post hoc test were employed for between-group comparisons of the data.

Results: AMPK and GLUT4 gene expression in the quadriceps muscle of the diabetic control group was significantly decreased compared to the healthy control group (P=0.0001). Moreover, the results showed that the content of AMPK and GLUT4 gene expression in diabetes + aerobic training and diabetes + aerobic training + berberine groups increased significantly compared to the diabetic control group (P=0.0001). This increase in gene expression was higher in the diabetes + aerobic training + berberine group than in the diabetes + aerobic training group. In addition, the level of AMPK and GLUT4 gene expression in the group of diabetes + aerobic training + berberine increased significantly compared to the diabetes + berberine group (P<0.05). However, despite the increase in gene expression of these two variables in diabetes + berberine group, these changes were not statistically significant (P>0.05) when compared to the diabetic control group.

Conclusion: The simultaneous intervention of aerobic training and berberine supplementation has positive synergistic effects on AMPK and GLUT4 gene expressions in the quadriceps muscle of type 1 diabetic rats and may improve energy homeostasis in the body.

Keywords

Main Subjects

biochemistry and metabolism

References

1. Han Y, Xiang Y, Shi Y, Tang X, Pan L, Gao J, et al. Pharmacokinetics and Pharmacological Activities of Berberine in Diabetes Mellitus Treatment. Evidence-based Complement Altern Med. 2021;2021.
2. Klip A, McGraw TE, James DE. Thirty sweet years of GLUT4. J Biol Chem. 2019;294(30):11369–81.
3. Foley KP, Klip A. Dynamic GLUT4 sorting through a syntaxin-6 compartment in muscle cells is derailed by insulin resistance-causing ceramide. Biol Open. 2014;3(5):314–25.
4. Li L V, Bakirtzi K, Watson RT, Pessin JE, Kandror K V. The C-terminus of GLUT4 targets the transporter to the perinuclear compartment but not to the insulin-responsive vesicles. Biochem J. 2009;419(1):105–13.
5. Shewan AM, van Dam EM, Martin S, Luen TB, Hong W, Bryant NJ, et al. GLUT4 recycles via a trans-Golgi network (TGN) subdomain enriched in Syntaxins 6 and 16 but not TGN38: involvement of an acidic targeting motif. Mol Biol Cell. 2003;14(3):973–86.
6. Xu Z, Kandror K V. Translocation of small preformed vesicles is responsible for the insulin activation of glucose transport in adipose cells: evidence from the in vitro reconstitution assay. J Biol Chem. 2002;277(50):47972–5.
7. Jedrychowski MP, Gartner CA, Gygi SP, Zhou L, Herz J, Kandror K V, et al. Proteomic analysis of GLUT4 storage vesicles reveals LRP1 to be an important vesicle component and target of insulin signaling. J Biol Chem. 2010;285(1):104–14.
8. Larance M, Ramm G, Stockli J, van Dam EM, Winata S, Wasinger V, et al. Characterization of the role of the Rab GTPase-activating protein AS160 in insulin-regulated GLUT4 trafficking. J Biol Chem. 2005;280(45):37803–13.
9. Karylowski O, Zeigerer A, Cohen A, McGraw TE. GLUT4 is retained by an intracellular cycle of vesicle formation and fusion with endosomes. Mol Biol Cell. 2004;15(2):870–82.
10. Schönke M, Myers MG, Zierath JR, Björnholm M. Skeletal muscle AMP-activated protein kinase γ1H151R overexpression enhances whole body energy homeostasis and insulin sensitivity. Am J Physiol - Endocrinol Metab. 2015;309(7):E679–90.
11. Bird SR, Hawley JA. Exercise and type 2 diabetes: new prescription for an old problem. Maturitas. 2012;72(4):311–6.
12. Chen Z-P, Stephens TJ, Murthy S, Canny BJ, Hargreaves M, Witters LA, et al. Effect of exercise intensity on skeletal muscle AMPK signaling in humans. Diabetes. 2003;52(9):2205–12.
13. Takekoshi K, Fukuhara M, Quin Z, Nissato S, Isobe K, Kawakami Y, et al. Long-term exercise stimulates adenosine monophosphate–activated protein kinase activity and subunit expression in rat visceral adipose tissue and liver. Metabolism. 2006;55(8):1122–8.
14. Cao S, Li B, Yi X, Chang B, Zhu B, Lian Z, et al. Effects of exercise on AMPK signaling and downstream components to PI3K in rat with type 2 diabetes. PLoS One. 2012;7(12):e51709.
15. Hall KE, McDonald MW, Grisé KN, Campos OA, Noble EG, Melling CWJ. The role of resistance and aerobic exercise training on insulin sensitivity measures in STZ-induced Type 1 diabetic rodents. Metabolism. 2013;62(10):1485–94.
16. Cunha VN, de Paula Lima M, Motta‐Santos D, Pesquero JL, de Andrade RV, de Almeida JA, et al. Role of exercise intensity on GLUT4 content, aerobic fitness and fasting plasma glucose in type 2 diabetic mice. Cell Biochem Funct. 2015;33(7):435–42.
17. Gurley JM, Griesel BA, Olson AL. Increased skeletal muscle GLUT4 expression in obese mice after voluntary wheel running exercise is posttranscriptional. Diabetes. 2016;65(10):2911–9.
18. NING G, HONG J, BI Y, GU W, ZHANG Y, ZHANG Z, et al. Progress in diabetes research in China. J Diabetes [Internet]. 2009 Sep;1(3):163–72.
19. Wu J, Li J, Li W, Sun B, Xie J, Cheng W, et al. Achyranthis bidentatae radix enhanced articular distribution and anti-inflammatory effect of berberine in Sanmiao Wan using an acute gouty arthritis rat model. J Ethnopharmacol. 2018;221:100–8.
20. Jin F, Xie T, Huang X, Zhao X. Berberine inhibits angiogenesis in glioblastoma xenografts by targeting the VEGFR2/ERK pathway. Pharm Biol. 2018;56(1):665–71.
21. Ikram M. A review on the chemical and pharmacological aspects of genus Berberis. Planta Med. 1975;28(08):353–8.
22. Vuddanda PR, Chakraborty S, Singh S. Berberine: a potential phytochemical with multispectrum therapeutic activities. Expert Opin Investig Drugs. 2010;19(10):1297–307.
23. Zhang W, XU Y, Guo F, Meng Y, Li M. Anti-diabetic effects of cinnamaldehyde and berberine and their impacts on retinol-binding protein 4 expression in rats with type 2 diabetes mellitus. Chin Med J (Engl). 2008;121(21):2124–8.
24. Wang Y, Campbell T, Perry B, Beaurepaire C, Qin L. Hypoglycemic and insulin-sensitizing effects of berberine in high-fat diet- and streptozotocin-induced diabetic rats. Metabolism. 2011 Feb;60(2):298–305.
25. Yeung AWK, Orhan IE, Aggarwal BB, Battino M, Belwal T, Bishayee A, et al. Berberine, a popular dietary supplement for human and animal health: Quantitative research literature analysis a review. 2020;
26. Zhou J, Zhou S, Tang J, Zhang K, Guang L, Huang Y, et al. Protective effect of berberine on beta cells in streptozotocin- and high-carbohydrate/high-fat diet-induced diabetic rats. Eur J Pharmacol [Internet]. 2009;606(1–3):262–8.
27. Chae CH, Jung SL, An SH, Park BY, Wang SW, Cho IH, et al. RETRACTED: Treadmill exercise improves cognitive function and facilitates nerve growth factor signaling by activating mitogen-activated protein kinase/extracellular signal-regulated kinase1/2 in the streptozotocin-induced diabetic rat hippocampus. Elsevier; 2009.
28. Arshadi S, Hasan Ghomi M, Banaeifar A.A, Kazemzadeh Y. The Effect of Eight Weeks Aerobic and Resistance Training on AMP-Activated Protein Kinase (AMPK) Gene Expression in Soleus Muscle and Insulin Resistance of STZ-Induced Diabetic Rat . jmciri 2019; 37 (2) :81-87. [In persian]
29. Eguchi T, Kumagai C, Fujihara T, Takemasa T, Ozawa T, Numata O. Black tea high-molecular-weight polyphenol stimulates exercise training-induced improvement of endurance capacity in mouse via the link between AMPK and GLUT4. PLoS One. 2013;8(7):e69480.
30. Chang W, Zhang M, Li J, Meng Z, Wei S, Du H, et al. Berberine improves insulin resistance in cardiomyocytes via activation of 5′-adenosine monophosphate-activated protein kinase. Metabolism. 2013;62(8):1159–67.
31. Turner N, Li J-Y, Gosby A, To SWC, Cheng Z, Miyoshi H, et al. Berberine and its more biologically available derivative, dihydroberberine, inhibit mitochondrial respiratory complex I: a mechanism for the action of berberine to activate AMP-activated protein kinase and improve insulin action. Diabetes. 2008;57(5):1414–8.
32. Xia X, Yan J, Shen Y, Tang K, Yin J, Zhang Y, et al. Berberine improves glucose metabolism in diabetic rats by inhibition of hepatic gluconeogenesis. PLoS One. 2011;6(2):1–10.
33. Shan Y, Zhang S, Gao B, Liang S, Zhang H, Yu X, et al. Adipose tissue SIRT1 regulates insulin sensitizing and anti-inflammatory effects of berberine. Front Pharmacol. 2020;11:591227.
34. Huang S, Czech MP. The GLUT4 glucose transporter. Cell Metab. 2007;5(4):237–52.
35. Astorino T, Baker J, Brock S, Dalleck L, Goulet E, Gotshall R, et al. Effect of exercise on GLUT4 expression of skeletal muscle in streptozotocin-induced diabetic rats. J Exerc Physiol. 2011;14(4).
36. Yin J, Ye J, Jia W. Effects and mechanisms of berberine in diabetes treatment. Acta Pharm Sin B [Internet]. 2012 Aug;2(4):327–34.
37. Zhang Q, Xiao X, Feng K, Wang T, Li W, Yuan T, et al. Berberine moderates glucose and lipid metabolism through multipathway mechanism. Evidence-Based Complement Altern Med. 2010;2011.
38. Wang Y-Y, Tang L-Q, Wei W. Berberine attenuates podocytes injury caused by exosomes derived from high glucose-induced mesangial cells through TGFβ1-PI3K/AKT pathway. Eur J Pharmacol. 2018;824:185–92.
39. Zhou T, Meng X, Che H, Shen N, Xiao D, Song X, et al. Regulation of insulin resistance by multiple MiRNAs via targeting the GLUT4 signalling pathway. Cell Physiol Biochem. 2016;38(5):2063–78.
40. Zhou D, Li S, Ding J, Yin T, Yang J, Ye H. MIF may participate in pathogenesis of polycystic ovary syndrome in rats through MAPK signalling pathway. Curr Med Sci. 2018;38(5):853–60.
41. Zhang N, Liu X, Zhuang L, Liu X, Zhao H, Shan Y, et al. Berberine decreases insulin resistance in a PCOS rats by improving GLUT4: Dual regulation of the PI3K/AKT and MAPK pathways. Regul Toxicol Pharmacol. 2020;110:104544.
42. Daimon M, Kamba A, Murakami H, Takahashi K, Otaka H, Makita K, et al. Association between pituitary-adrenal axis dominance over the renin-angiotensin-aldosterone system and hypertension. J Clin Endocrinol Metab. 2016;101(3):889–97.
43. Perry RJ, Zhang X-M, Zhang D, Kumashiro N, Camporez J-PG, Cline GW, et al. Mechanism for the anti-diabetic effect of leptin. Nat Med. 2014;20(7):759.
44. DIMITRIADIS G, LEIGHTON B, PARRY-BILLINGS M, SASSON S, YOUNG M, KRAUSE U, et al. Effects of glucocorticoid excess on the sensitivity of glucose transport and metabolism to insulin in rat skeletal muscle. Biochem J. 1997;321(3):707–12.
45. Piroli GG, Grillo CA, Reznikov LR, Adams S, McEwen BS, Charron MJ, et al. Corticosterone impairs insulin-stimulated translocation of GLUT4 in the rat hippocampus. Neuroendocrinology. 2007;85(2):71–80.
46. Mi J, He W, Lv J, Zhuang K, Huang H, Quan S. Effect of berberine on the HPA-axis pathway and skeletal muscle GLUT4 in type 2 diabetes mellitus rats. Diabetes, Metab Syndr Obes. 2019;12:1717–25.