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水提取物处理的HepG2细胞中葡萄糖代谢增加及α-葡萄糖苷酶抑制作用

Increased glucose metabolism and alpha-glucosidase inhibition in water extract-treated HepG2 cells.

作者信息

Kim Dae Jung, Kang Yun Hwan, Kim Kyoung Kon, Kim Tae Woo, Park Jae Bong, Choe Myeon

机构信息

Well-being Bioproducts RIC, Kangwon National University, Gangwon 25209, Korea.

National Development Institute of Korean Medicine, Gyeongbuk 38540, Korea.

出版信息

Nutr Res Pract. 2017 Jun;11(3):180-189. doi: 10.4162/nrp.2017.11.3.180. Epub 2017 May 22.

Abstract

BACKGROUND/OBJECTIVES: Recent living condition improvements, changes in dietary habits, and reductions in physical activity are contributing to an increase in metabolic syndrome symptoms including diabetes and obesity. Through such societal developments, humankind is continuously exposed to metabolic diseases such as diabetes, and the number of the victims is increasing. This study investigated water extract (CMW)-induced glucose uptake in HepG2 cells and the effect of CMW treatment on glucose metabolism.

MATERIALS/METHODS: Colorimetric assay kits were used to determine the glucokinase (GK) and pyruvate dehydrogenase (PDH) activities, glucose uptake, and glycogen content. Either RT-PCR or western blot analysis was performed for quantitation of glucose transporter 2 (GLUT2), hepatocyte nuclear factor 1 alpha (HNF-1α), phosphatidylinositol 3-kinase (PI3k), protein kinase B (Akt), phosphorylated AMP-activated protein kinase (pAMPK), phosphoenolpyruvate carboxykinase, GK, PDH, and glycogen synthase kinase 3 beta (GSK-3β) expression levels. The α-glucosidase inhibitory activities of acarbose and CMW were evaluated by absorbance measurement.

RESULTS

CMW induced glucose uptake in HepG2 cells by increasing GLUT2 through HNF-1α expression stimulation. Glucose in the cells increased the CMW-induced phosphorylation of AMPK. In turn, glycolysis was stimulated, and glyconeogenesis was inhibited. Furthermore, by studying the mechanism of action of PI3k, Akt, and GSK-3β, and measuring glycogen content, the study confirmed that the glucose was stored in the liver as glycogen. Finally, CMW resulted in a higher level of α-glucosidase inhibitory activity than that from acarbose.

CONCLUSION

CMW induced the uptake of glucose into HepG2 cells, as well, it induced metabolism of the absorbed glucose. It is concluded that CMW is a candidate or potential use in diabetes prevention and treatment.

摘要

背景/目的:近期生活条件的改善、饮食习惯的改变以及体力活动的减少,导致包括糖尿病和肥胖症在内的代谢综合征症状有所增加。随着这些社会发展,人类不断受到糖尿病等代谢性疾病的影响,患者数量也在增加。本研究调查了朝鲜淫羊藿水提取物(CMW)诱导HepG2细胞摄取葡萄糖的情况以及CMW处理对葡萄糖代谢的影响。

材料/方法:使用比色测定试剂盒来测定葡萄糖激酶(GK)和丙酮酸脱氢酶(PDH)的活性、葡萄糖摄取量以及糖原含量。采用逆转录聚合酶链反应(RT-PCR)或蛋白质免疫印迹分析(western blot analysis)对葡萄糖转运蛋白2(GLUT2)、肝细胞核因子1α(HNF-1α)、磷脂酰肌醇3激酶(PI3k)、蛋白激酶B(Akt)、磷酸化的腺苷酸活化蛋白激酶(pAMPK)、磷酸烯醇式丙酮酸羧激酶、GK、PDH和糖原合酶激酶3β(GSK-3β)的表达水平进行定量分析。通过吸光度测量评估阿卡波糖和CMW的α-葡萄糖苷酶抑制活性。

结果

CMW通过刺激HNF-1α表达增加GLUT2,从而诱导HepG2细胞摄取葡萄糖。细胞内的葡萄糖增加了CMW诱导的AMPK磷酸化。进而刺激糖酵解并抑制糖异生。此外,通过研究PI3k、Akt和GSK-3β的作用机制并测量糖原含量,该研究证实葡萄糖以糖原的形式储存在肝脏中。最后,CMW的α-葡萄糖苷酶抑制活性水平高于阿卡波糖。

结论

CMW诱导葡萄糖进入HepG2细胞,同时也诱导了所吸收葡萄糖的代谢。得出结论,CMW是预防和治疗糖尿病的候选药物或具有潜在用途。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/5449374/681e21651d16/nrp-11-180-g001.jpg

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