淫羊藿通过miR-19a-3p对高糖高脂诱导的胰岛β细胞损伤的影响及机制研究

陈朝琴; 薛治乾; 李文霞

doi:10.13898/j.cnki.issn.1000-2200.2020.06.007

淫羊藿通过miR-19a-3p对高糖高脂诱导的胰岛β细胞损伤的影响及机制研究

Effects of Epimedium herb through miR-19a-3p on islet β cells damage induced by high glucose and lipid and its mechanism

摘要

摘要:
目的探讨淫羊藿（Epimedium herb，EH）对高糖高脂诱导的胰岛β细胞损伤的影响及其分子机制。
方法以胰岛β细胞RINm5F为研究对象，建立高糖高脂损伤模型，分为对照组（5 mmol/L葡萄糖）、高糖高脂组25 mmol/L葡萄糖+0.25 mmol/L棕榈树酸（PA）、低剂量EH组（25 mmol/L葡萄糖+0.25 mmol/L PA+10 μmol/L EH）和高剂量EH组（25 mmol/L葡萄糖+0.25 mmol/L PA+100 μmol/L EH），EH处理48 h后，分别采用噻唑蓝（MTT）比色法和流式细胞术检测细胞增殖与凋亡，蛋白质印迹法（Western blotting）检测细胞中细胞周期蛋白D1（CyclinD1）、P21、B细胞淋巴瘤/白血病-2（Bcl-2）和Bcl-2相关X蛋白（Bax）蛋白表达水平，实时定量PCR（qPCR）检测RINm5F细胞中miR-19a-3p表达。在高糖高脂损伤的RINm5F细胞转染anti-miR-19a-3p或在高剂量EH组细胞转染miR-19a-3p，观察细胞的增殖和凋亡情况。
结果与对照组比较，高糖高脂组第48 h、72 h细胞增殖活性和CyclinD1、Bcl-2蛋白水平明显下降（P < 0.05），细胞凋亡率、miR-19a-3p表达量、P21和Bax蛋白表达量提高（P < 0.05）。不同剂量的EH干预可以明显提高高糖高脂环境下的细胞增殖能力和CyclinD1、Bcl-2蛋白水平（P < 0.01），减少细胞凋亡率和P21、Bax蛋白表达量（P < 0.01），与抑制miR-19a-3p表达作用结果差异无统计学意义（P>0.05）。此外，过表达miR-19a-3p能逆转EH对高糖高脂刺激的RINm5F细胞增殖的促进作用和对细胞凋亡的抑制作用（P < 0.01）。
结论EH通过miR-19a-3p保护高糖高脂诱导的胰岛β细胞损伤，提高细胞增殖活性，抑制细胞凋亡。

Abstract:
ObjectiveTo investigate the effects and molecular mechanism of Epimedium herb(EH) on islet β cells injury induced by high glucose and lipid.
MethodsThe model of high glucose and lipid injury of islet β cells RINm5F was established.The cells were divided into the control group(treatment with 5 mmol/L glucose), high glucose and lipid(treatment with 25 mmol/L glucose and 0.25 mmol/L PA), low dose EH group(treatment with 25 mmol/L glucose, 0.25 mmol/L PA and 10 μmol/L EH), high dose EH group(treatment with 25 mmol/L glucose, 0.25 mmol/L PA and 100 μmol/L EH).After 48 h of treatment, the cell proliferation and apoptosis were detected using MTT assay and flow cytometry, respectively.The expression levels of CyclinD1, P21, B cell lymphoma/lewkmia-2(Bcl-2) and Bcl-2 associated X(Bax) protein were analyzed using Western blotting.The expression level of miR-19a-3p mRNA in RINm5F cells was detected using real-time quantitative PCR(qPCR).The high glucose and lipid damaged RINm5F cells were transfected using anti-miR-19a-3p or miR-19a-3p in the high dose EH group, and the proliferation and apoptosis of the cells were observed.
ResultsCompared with the control group, the cell proliferation activities at 48 h and 72 h, CyclinD1 and Bcl-2 protein levels significantly decreased(P < 0.05), and the apoptosis rate, miR-19a-3p expression, P21 and Bax protein expression levels significantly increased in high glucose and lipid group(P < 0.05).The intervention of different doses of EH could significantly improve the proliferation capacity and levels of CyclinD1 and Bcl-2 protein of cells under high glucose and lipid environment, and significantly reduced the apoptosis rate and expression levels of P21 and Bax protein(P < 0.01), and the difference between which and effect of inhibiting miR-19a-3p was not statistically significant(P>0.05).The overexpression of miR-19a-3p could reverse the EH promoting effect on proliferation and inhibiting apoptosis of RINm5F cells stimulated by high glucose and lipid(P < 0.01).
ConclusionsEH protects islet β cells damage induced by high glucose and lipid through miR-19a-3P, improves cell proliferation activity and inhibits apoptosis.

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