论著 Original Article

Wnt/β-catenin信号在高糖诱导H9c2心肌细胞损伤与凋亡中的作用

Published at: 2016年第36卷第12期

王记 1 , 陆玉琴 1 , 赵信科 1 , 纪召娟 1
1 甘肃中医药大学附属医院心血管科,兰州 730020
通讯作者 记 王 Email: doctorwangji@163.com
DOI: 10.3978/j.issn.2095-6959.2016.12.012
基金:
甘肃省科技厅自然科学基金 1508RJZA044

摘要

目的:探讨Wnt/β-catenin信号通路是否参与体外培养条件下高糖诱导的H9c2心肌细胞损伤凋亡,及其与细胞自噬的关系。方法:构建高糖诱导H9c2心肌细胞凋亡细胞模型,CCK-8法和流式细胞仪检测细胞存活率和凋亡率,Western blot检测H9c2心肌细胞caspase-3、LC3-Ⅱ/LC3-Ⅰ、Axin-1和β-catenin的表达。结果:与对照组相比,50 mmol/L高糖处理H9c2心肌细胞48 h后,细胞存活率急速下降,caspase-3表达增强;其次,Western blot实验结果显示高糖促进wnt/β-catenin的激活,表现为Axin-1表达抑制,β-catenin表达显著升高,并伴随细胞内LC3-Ⅱ/LC3-Ⅰ比值降低;DKK预处理能削弱高糖引起的H9c2心肌细胞凋亡,下调Axin-1活性,增强β-catenin表达和LC3-Ⅱ/LC3-Ⅰ比值。结论:Wnt/β-catenin通路在高糖致H9c2心肌细胞损伤与凋亡过程中被激活,且其的激活抑制了自噬通路的活化。而当添加D-葡萄糖(Dickkopf-1,DKK-1)抑制Wnt/β-catenin后,自噬作用增强,高糖诱导的H9c2心肌细胞凋亡率明显下降。


Role of Wnt/β-catenin pathway on high glucose-induced H9c2 myocardial cell injury and apoptosis

Abstract

Objective: To investigate role of Wnt/β-catenin pathway on high glucose-induced H9c2 myocardial cell apoptosis, and evaluate the relationship between Wnt/β-catenin and autophagy. Methods: H9c2 myocardial cell apoptosis model induced by high glucose was constructed, cell viability and apoptosis ratio was determined by CCK-8 and flow cytometry, Western blot analysis was used to detect the expression of caspase-3, LC3-Ⅱ/LC3-Ⅰ, Axin-1 and β-catenin in H9c2 myocardial cell. Results: Cell viability fell sharply, and protein expression of caspase-3 were remarkably increased in H9c2 myocardial cell treatment with 50 mmol/L high glucose for 48 h compared with control cells. Furthermore, Western blot analysis revealed that high glucose triggered Wnt pathways, thus suppressed the expression level of Axin-1 and promoted β-catenin expression and LC3-Ⅱ/LC3-Ⅰ ratio. Pretreatment by dickkopf-1(DKK-1) attenuated HG-induced apoptosis of H9c2 myocardial cell, down-regulated the activity of Axin-1, and significantly increased the expression of β-catenin and LC3-Ⅱ/LC3-Ⅰ ratio, and reduced the production of β-catenin. Conclusion: This study has demonstrated that Wnt/β-catenin pathway was activated on HG-induced injury and apoptosis in H9c2 cells and that was associated with inhibition of autophagy pathways. Furthermore, blocking the Wnt/β-catenin signaling with the selective antagonist DKK-1 resulted in enhancement of autophagy, and inhibition of the apoptotic index in high glucose-induced H9c2 myocardial cell.


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引用

引用本文: 记 王, 玉琴 陆, 信科 赵, 召娟 纪. Wnt/β-catenin信号在高糖诱导H9c2心肌细胞损伤与凋亡中的作用[J]. 临床与病理杂志, 2016, 36(12): 1955-1960.
Cite this article as: WANG Ji, LU Yuqin, ZHAO Xinke, JI Zhaojuan . Role of Wnt/β-catenin pathway on high glucose-induced H9c2 myocardial cell injury and apoptosis[J]. Journal of Clinical and Pathological Research, 2016, 36(12): 1955-1960.