肾小球系膜细胞中Β_1整合素表达及细胞凋亡与N-糖链抑制剂关系的研究

Effects of N-linked Oligosaccharide Chains on Expression of Integrin β_1 and Apoptosis in Glomerular Mesangial Cells

目的观察N-糖链抑制剂衣霉素(TM)对肾小球系膜细胞(GMC)增殖、凋亡的影响以及Β1整合素和CYCLIN D1表达在细胞凋亡以及细胞增殖中的作用。方法体外培养的大鼠GMC分四组:正常对照组,0.5、1.0和2.0ΜG/ML衣霉素组。采用流式细胞技术测定Β1整合素水平和细胞凋亡率,细胞免疫化学方法测定CYCLIN D1表达,MTT法测定细胞增殖能力。结果TM作用以后,细胞Β1整合素水平降低,细胞凋亡率增加,同时细胞周期正调控蛋白CYCLIN D1表达下降,增殖能力降低,这种作用呈剂量依赖性。结论N-糖链抑制剂TM通过抑制Β1整合素的表达,影响细胞的粘附而导致细胞凋亡,并影响细胞周期正调控蛋白CYCLIN D1表达,使细胞增殖能力受抑,这种作用呈剂量依赖性。

Objective The effects of N-linked oligosaccharide chains（TM）, an inhibitor of N-glycosylation of proteins, on the expression of β1 integrin and the apoptosis of glomerular mesangial cells（GMC） were observed for exploring the role of inhibitors of N-linked oligosaccharide chains in cell apoptosis, proliferation, and expression of β1 integrin and cyclin D1, and hence finding a new approach to the synteresis of proliferative nephropathy. Methods Cultured rat mesangial cells were divided into 4 groups： control group, and 0. 5, 1.0 and 2. 0 μg/mL TM-treatment groups. The expression of β1 integrin and apoptosis rate were measured by flow cytometry; the expression of cyclin D1 was measured by immunobistocbemistry, and proliferation of GMCs was measured by MTT. Results The expression of β1 integrin and cyclin D1 were decreased notably by inhibitors of N-linked oligosaccharide chains-TM. TM increased apoptosis and decreased the proliferative abilitiy of cells, and all the effects of TM were dose-dependent. Conclusion Through repressed glycosylation of glycoprotein, TM can suppress the expression of β1 integrin, affect the adhesion of cells, and increase the apoptosis rate; at the same time, the expression of cyclin DI and the proliferative ability of cells were decreased, and all these were dosedependent.