心肌成纤维细胞Wnt信号通路在高糖环境下的调控变化

Regulation Change on Wnt Signal Pathway of Cardiac Fibroblasts in High Glucose Environment

目的:观察乳鼠心肌成纤维细胞在高浓度葡萄糖作用下胶原蛋白分泌和细胞增殖及Wnt信号通路调控的变化。方法:基于高糖刺激诱导心肌成纤维细胞体外增殖的培养方法,CCK8法检测细胞增殖能力;流式细胞仪检测细胞的增殖周期;ELISA法观察细胞TGF-β1蛋白及I型、Ⅲ型胶原的分泌能力;western blot法检测蛋白β-catenin,GSK-3β,P-GSK-3β的表达水平;观察心肌成纤维细胞在高糖环境中Wnt信号通路传导的变化。结果:高浓度葡萄糖环境可以引起细胞的增殖(P<0.01);促进TGF-β1的合成(P<0.01);提高细胞I型、Ⅲ型胶原分泌的能力(P<0.01);在分子水平上可以增强β-catenin的表达(P<0.01),抑制P-GSK-3β的表达(P<0.05)。结论:心肌成纤维细胞在高糖环境中增殖明显,心肌成纤维细胞TGF-β1和胶原的分泌会增加,引发Wnt信号通路的异常表达。研究其作用机制,对糖尿病心肌纤维化的防治及研究具有重大意义。

Objective： Observe the rats cardiac fibroblasts proliferation and protein secretion and Wnt signal pathway regulation change in the high concentration glucose environment. Methods： Based on the high glucose stimulation induced cardiac fibroblasts proliferation in vitro, CCK8 method to detect the cell proliferation; flow cytometry detect the cell cycle; ELISA method to observe the ability of the cell TGF-β1, type I and type III collagen protein secretion; western blot method detection the expression level of β-catenin, GSK-3β, P-GSK-3β ; to observe the variation of high glucose environment on cardiac fibrobtasts of Wnt signal pathway conduction. Results： High concentration glucose environment can cause cell proliferation （P 〈 0. 01 ） ; improve the ability of type I and type III collagen secretion （P 〈 0. 01 ） ; promote the synthesis of TGF-β, （ P 〈 0. 01 ） ; upregulate the expression of β-catenin （ P 〈 0. 01）, reduce the expression of P-GSK-3β （ P 〈 0. 05 ） at the molecular level. Conclusion： The cardiac fibroblasts can obviously proliferation in high glucose environment, the cardiac fibroblasts TGF-β1 and collagen secretion can increased, cause abnormal expression of Wnt signaling pathway. Research its mechanism of action, have the great significance for prevention and treatment and research diabetic myocardial fibrosis.