Effect of oxymatrine on liver gluconeogenesis is associated with the regulation of PEPCK and G6Pase expression and AKT phosphorylation

Zhu,Yu-Xian; Hu,Hai-Qing; Zuo,Mei-Ling; Mao,Li; Song,Gui-Lin; Li,Tao-Ming; Dong,Li-Chen; Yang,Zhong-Bao; Ali Sheikh,Md Sayed

doi:10.3892/br.2021.1432

Effect of oxymatrine on liver gluconeogenesis is associated with the regulation of PEPCK and G6Pase expression and AKT phosphorylation

Authors:
- Yu-Xian Zhu
- Hai-Qing Hu
- Mei-Ling Zuo
- Li Mao
- Gui-Lin Song
- Tao-Ming Li
- Li-Chen Dong
- Zhong-Bao Yang
- Md Sayed Ali Sheikh
View Affiliations

Affiliations: The Affiliated Changsha Hospital of Hunan Normal University, Changsha, Hunan 410006, P.R. China, Department of Basic Medicine, Changsha Health Vocational College, Changsha, Hunan 410600, P.R. China, Internal Medicine Department, Cardiology, College of Medicine, Al Jouf University, Sakaka, Al Jouf 72388, Saudi Arabia
Published online on: April 27, 2021 https://doi.org/10.3892/br.2021.1432
Article Number: 56
Copyright: © Zhu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

An increase in liver gluconeogenesis is an important pathological phenomenon in type 2 diabetes mellitus (T2DM) and oxymatrine is an effective natural drug used for T2DM treatment. The present study aimed to explore the effect of oxymatrine on gluconeogenesis and elucidate the underlying mechanism. Male Sprague‑Dawley rats were treated with a high‑fat diet and streptozotocin for 4 weeks to induce T2DM, and HepG2 cells were treated with 55 mM glucose to simulate T2DM in vitro. T2DM rats were treated with oxymatrine (10 or 20 mg/kg weight) or metformin for 4 weeks, and HepG2 cells were treated with oxymatrine (0.1 or 1 µM), metformin (0.1 µM), or oxymatrine combined with MK‑2206 (AKT inhibitor) for 24 h. Fasting blood glucose and insulin sensitivity of rats were measured to evaluate insulin resistance. Glucose production and uptake ability were measured to evaluate gluconeogenesis in HepG2 cells, and the expression of related genes was detected to explore the molecular mechanism. Additionally, the body weight, liver weight and liver index were measured and hematoxylin and eosin staining was performed to evaluate the effects of the disease. The fasting glucose levels of T2DM rats was 16.5 mmol/l, whereas in the control rats, it was 6.1 mmol/l. Decreased insulin sensitivity (K‑value, 0.2), body weight loss (weight, 300 g), liver weight gain, liver index increase (value, 48) and morphological changes were observed in T2DM rats, accompanied by reduced AKT phosphorylation, and upregulated expression of phosphoenolpyruvate carboxykinase (PEPCK) and glucose‑6‑phosphatase (G6Pase). High‑glucose treatment significantly increased glucose production and decreased glucose uptake in HepG2 cells, concomitant with a decrease in AKT phosphorylation and increase of PEPCK and G6Pase expression. In vivo, oxymatrine dose‑dependently increased the sensitivity of T2DM rats to insulin, increased AKT phosphorylation and decreased PEPCK and G6Pase expression in the liver, and reversed the liver morphological changes. In vitro, oxymatrine dose‑dependently increased AKT phosphorylation and glucose uptake of HepG2 cells subjected to high‑glucose treatment, which was accompanied by inhibition of the expression of the gluconeogenesis‑related genes, PEPCK and G6Pase. MK‑2206 significantly inhibited the protective effects of oxymatrine in high‑glucose‑treated cells. These data indicated that oxymatrine can effectively prevent insulin resistance and gluconeogenesis, and its mechanism may be at least partly associated with the regulation of PEPCK and G6Pase expression and AKT phosphorylation in the liver.

View Figures

View References

1	American Diabetes Association. Diagnosis and classification of diabetes mellitus. Diabetes Care. 37 (Suppl 1):S81–S90. 2014.PubMed/NCBI View Article : Google Scholar
2	Ma RCW: Epidemiology of diabetes and diabetic complications in China. Diabetologia. 61:1249–1260. 2018.PubMed/NCBI View Article : Google Scholar
3	Wu Y, Ding Y, Tanaka Y and Zhang W: Risk factors contributing to type 2 diabetes and recent advances in the treatment and prevention. Int J Med Sci. 11:1185–1200. 2014.PubMed/NCBI View Article : Google Scholar
4	Rines AK, Sharabi K, Tavares CD and Puigserver P: Targeting hepatic glucose metabolism in the treatment of type 2 diabetes. Nat Rev Drug Discov. 15:786–804. 2016.PubMed/NCBI View Article : Google Scholar
5	Rui L: Energy metabolism in the liver. Compr Physiol. 4:177–197. 2014.PubMed/NCBI View Article : Google Scholar
6	De Meyts P: The insulin receptor and its signal transduction network. [Updated 2016 Apr 27]. In. Feingold KR, Anawalt B, Boyce A, Chrousos G, de Herder WW, Dhatariya K, Dungan K, Grossman A, Hershman JM, Hofland J, et al (eds). Endotext (Internet). South Dartmouth (MA), MDText.com, Inc., 2000. https://www.ncbi.nlm.nih.gov/books/NBK378978. Accessed April 27, 2016.
7	Muoio DM and Newgard CB: Mechanisms of disease: Molecular and metabolic mechanisms of insulin resistance and beta-cell failure in type 2 diabetes. Nat Rev Mol Cell Biol. 9:193–205. 2008.PubMed/NCBI View Article : Google Scholar
8	Fan H, Li L, Zhang X, Liu Y, Yang C, Yang Y and Yin J: Oxymatrine downregulates TLR4, TLR2, MyD88 and NF-kappa B and protects rat brains against focal ischemia. Mediators Inflamm. 2009(704706)2009.PubMed/NCBI View Article : Google Scholar
9	Liu Y, Xu Y, Ji W, Li X, Sun B, Gao Q and Su C: Anti-tumor activities of matrine and oxymatrine: Literature review. Tumour Biol. 35:5111–5119. 2014.PubMed/NCBI View Article : Google Scholar
10	Chen XS, Wang GJ, Cai X, Yu HY and Hu YP: Inhibition of hepatitis B virus by oxymatrine in vivo. World J Gastroenterol. 7:49–52. 2001.PubMed/NCBI View Article : Google Scholar
11	Wu XL, Zeng WZ, Jiang MD, Qin JP and Xu H: Effect of oxymatrine on the TGF beta-Smad signaling pathway in rats with CCl4-induced hepatic fibrosis. World J Gastroenterol. 14:2100–2105. 2008.PubMed/NCBI View Article : Google Scholar
12	Chan SM and Ye JM: Strategies for the discovery and development of anti-diabetic drugs from the natural products of traditional medicines. J Pharm Pharm Sci. 16:207–216. 2013.PubMed/NCBI View Article : Google Scholar
13	Wang L, Li X, Zhang Y, Huang Y, Zhang Y and Ma Q: Oxymatrine ameliorates diabetes-induced aortic endothelial dysfunction via the regulation of eNOS and NOX4. J Cell Biochem, Nov 19, 2018 (Epub ahead of print). doi: 10.1002/jcb.28006.
14	Wang SB and Jia JP: Oxymatrine attenuates diabetes-associated cognitive deficits in rats. Acta Pharmacol Sin. 35:331–338. 2014.PubMed/NCBI View Article : Google Scholar
15	Lu ML, Xiang XH and Xia SH: Potential signaling pathways involved in the clinical application of oxymatrine. Phytother Res. 30:1104–1112. 2016.PubMed/NCBI View Article : Google Scholar
16	National Research Council (US) Committee for the update of the Guide for the Care and Use of Laboratory Animals: Guide for the Care and Use of Laboratory Animals. 8th edition. National Academicals Press, Washington, DC, 2011.
17	Ma C, Yu H, Xiao Y and Wang H: Momordica charantia extracts ameliorate insulin resistance by regulating the expression of SOCS-3 and JNK in type 2 diabetes mellitus rats. Pharm Biol. 55:2170–2177. 2017.PubMed/NCBI View Article : Google Scholar
18	Zuo ML, Wang AP, Tian Y, Mao L, Song GL and Yang ZB: Oxymatrine ameliorates insulin resistance in rats with type 2 diabetes by regulating the expression of KSRP, PETN, and AKT in the liver. J Cell Biochem. 120:16185–16194. 2019.PubMed/NCBI View Article : Google Scholar
19	Yoshioka K, Saito M, Oh KB, Nemoto Y, Matsuoka H, Natsume M and Abe H: Intracellular fate of 2-NBDG, a fluorescent probe for glucose uptake activity, in Escherichia coli cells. Biosci Biotechnol Biochem. 60:1899–1901. 1996.PubMed/NCBI View Article : Google Scholar
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
21	Zdychova J and Komers R: Emerging role of AKT kinase/protein kinase B signaling in pathophysiology of diabetes and its complications. Physiol Res. 54:1–16. 2005.PubMed/NCBI
22	Huang X, Liu G, Guo J and Su Z: The PI3K/AKT pathway in obesity and type 2 diabetes. Int J Biol Sci. 14:1483–1496. 2018.PubMed/NCBI View Article : Google Scholar
23	Vinayagam A, Kulkarni MM, Sopko R, Sun X, Hu Y, Nand A, Villalta C, Moghimi A, Yang X, Mohr SE, et al: An integrative analysis of the InR/PI3K/AKT network identifies the dynamic response to insulin signaling. Cell Rep. 16:3062–3074. 2016.PubMed/NCBI View Article : Google Scholar
24	Bathina S and Das UN: Dysregulation of PI3K-AKT-mTOR pathway in brain of streptozotocin-induced type 2 diabetes mellitus in Wistar rats. Lipids Health Dis. 17(168)2018.PubMed/NCBI View Article : Google Scholar
25	Burgess SC, Hausler N, Merritt M, Jeffrey FM, Storey C, Milde A, Koshy S, Lindner J, Magnuson MA, Malloy CR and Sherry AD: Impaired tricarboxylic acid cycle activity in mouse livers lacking cytosolic phosphoenolpyruvate carboxykinase. J Biol Chem. 279:48941–48949. 2004.PubMed/NCBI View Article : Google Scholar
26	Mutel E, Abdul-Wahed A, Ramamonjisoa N, Stefanutti A, Houberdon I, Cavassila S, Pilleul F, Beuf O, Gautier-Stein A, Penhoat A, et al: Targeted deletion of liver glucose-6 phosphatase mimics glycogen storage disease type l a including development of multiple adenomas. J Hepatol. 54:529–537. 2011.PubMed/NCBI View Article : Google Scholar
27	Jiang WJ, Wang S, Xiao M, Lin Y, Zhou L, Lei Q, Xiong Y, Guan KL and Zhao S: Acetylation regulates gluconeogenesis by promoting PEPCK1 degradation via recruiting the UBR5 ubiquitin ligase. Mol Cell. 43:33–44. 2011.PubMed/NCBI View Article : Google Scholar
28	Koo SH, Flechner L, Qi L, Zhang X, Screaton RA, Jeffries S, Hedrick S, Xu W, Boussouar F, Brindle P, et al: The CREB coactivator TORC2 is a key regulator of fasting glucose metabolism. Nature. 437:1109–1111. 2005.PubMed/NCBI View Article : Google Scholar
29	Matsumoto M, Pocai A, Rossetti L, Depinho RA and Accili D: Impaired regulmion of hepatic glucose production in mice lacking the forkhead transcription factor Foxol in liver. Cell Metab. 6:208–216. 2007.PubMed/NCBI View Article : Google Scholar
30	Zhou XY, Shibusawa N, Naik K, Porras D, Temple K, Ou H, Kaihara K, Roe MW, Brady MJ and Wondisford FE: Insulin regulation of hepatic gluconeogenesis through phosphorylation of CREB-binding protein. Nat Med. l0:633–637. 2004.PubMed/NCBI View Article : Google Scholar
31	Erion DM, Ignatova ID, Yonemitsu S, Nagai Y, Chatterjee P, Weismann D, Hsiao JJ, Zhang D, 1wasaki T, Stark R, et al: Prevention of hepatic steatosis and hepatic insulin resistance by knockdown of cAMP response element-binding protein. Cell Metab. 10:499–506. 2009.PubMed/NCBI View Article : Google Scholar
32	Cypess AM, Zhang H, Schulz TJ, Huang TL, Espinoza DO, Kristiansen K, Unterman TG and Tseng YH: Insulin/IGF-I regulation of necdin and brown adipocyte differentiation via CREB- and FoxO1-associated pathways. Endocrinology. 152:3680–3689. 2011.PubMed/NCBI View Article : Google Scholar
33	Li X, Monks B, Ge Q and Birnbaum MJ: AKT/PKB regulates hepatic metabolism by directly inhibiting PGC-1alpha transcription coactivator. Nature. 447:1012–1016. 2007.PubMed/NCBI View Article : Google Scholar
34	Abdul-Ghani MA, Puckett C, Triplitt C, Maggs D, Adams J, Cersosimo E and DeFronzo RA: Initial combination therapy with metformin, pioglitazone and exenatide is more effective than sequential add-on therapy in subjects with new-onset diabetes. Results from the efficacy and durability of initial combination therapy for Type 2 Diabetes (EDICT): A randomized trial. Diabetes Obes Metab. 17:268–275. 2015.PubMed/NCBI View Article : Google Scholar
35	Guo C, Zhang C, Li L, Wang Z, Xiao W and Yang Z: Hypoglycemic and hypolipidemic effects of oxymatrine in high-fat diet and streptozotocin-induced diabetic rats. Phytomedicine. 21:807–814. 2014.PubMed/NCBI View Article : Google Scholar
36	Yu XH, Zhu JS and Gu GM: Clinical effect of oxymatrine combined with metformin on insulin resistance and serum TNF-α in patients with non-alcoholic fatty liver. Chin J Clin Hepatol. 23:195. 2007.