活性维生素D_3对人系膜细胞凋亡及Caspase-3表达的影响

Effect of 1,25(OH)_2D_3 on Apoptosis of Human Mesangial Cells and Caspase-3 Expression

目的探讨活性维生素D3〔1,25(OH)2D3〕对人系膜细胞凋亡的影响,并初步探究其作用机制。方法体外培养人系膜细胞,随机分为4组:正常对照组,表皮生长因子(EGF)组,1,25(OH)2D3组,EGF联合1,25(OH)2D3组,采用流式细胞术检测各组系膜细胞的凋亡率,Western blotting检测各组半胱氨酸蛋白酶-3(Caspase-3)蛋白表达的情况。结果与正常对照组比较,EGF组系膜细胞凋亡率降低,Caspase-3表达量降低(P<0.05),1,25(OH)2D3组系膜细胞的凋亡率升高,Caspase-3表达量升高(P<0.05);与EGF组比较,EGF联合1,25(OH)2D3组系膜细胞的凋亡率升高,Caspase-3表达量升高(P<0.05);EGF联合1,25(OH)2D3组与正常对照组系膜细胞的凋亡率和Caspase-3表达量间差异均无统计学意义(P>0.05)。结论 1,25(OH)2D3可能是通过上调Caspase-3的表达而诱导系膜细胞的凋亡。EGF可抑制人系膜细胞的凋亡,1,25(OH)2D3可逆转EGF对系膜细胞凋亡的抑制作用。

Objective To investigate the effect of 1,25- dihydroxyvitamin D3〔1,25（ OH）2D3〕on apoptosis of human mesangial cells and its mechanism. Methods The mesangial cells cultured in vitro were divided randomly into groups A（ normal control group）,B 〔epidermal growth factor（ EGF） group〕,C 〔1,25（ OH）2D3 group〕,D 〔combined group of EGF and 1,25（ OH）2D3〕. Flow cytometry was used to determine the apoptosis rates,the Western blotting to detect the expression of cysteine proteinase- 3（ caspase- 3） in 4 groups. Results As compared with group A, the apoptosis rate of mesangial cells decreased,caspase- 3 expression reduced in group B（ P〈0. 05）,the apoptosis rate of mesangial cells and caspase- 3 expression increased in group C（ P〈0. 05）. As compared with group B,the apoptosis rate of mesangial cells and caspase- 3 expression elevated in group D（ P〈0. 05）,there was no significant difference between groups A,D（ P〈0. 05）.Conclusion 1,25（ OH）2D3 may induce the apoptosis of mesangial cells through up- regulating casoase- 3 expression. EGF inhibits the apoptosis,1,25（ OH）2D3 can reverse the inhibition of EGF on mesangial cells.