高浓度葡萄糖损伤人腹膜间皮细胞分子机制的研究

The study on molecular mechanism of high-glucose-inducing injury to human peritoneal mesothelial cells

目的:探讨高浓度葡萄糖损伤人腹膜间皮细胞的机制。方法:(1)人腹膜间皮细胞分别经含1.5%、2.5%、4.25%葡萄糖的M199培养基培养48小时后,检测间皮细胞线粒体膜电位(Ψ)、凋亡率、caspase-3活性、bax和bcl-2基因mRNA表达;(2)分别用0.1μmol/L和0.01μmol/L环孢素A(CsA)与4.25%葡萄糖共同作用人腹膜间皮细胞,检测细胞凋亡率和caspase-3活性。结果:(1)随着葡萄糖浓度增加,Ψ丧失的细胞百分率、bax mRNA表达量、caspase-3活性、凋亡率增加(P<0.05),而bcl-2 mRNA表达量减少;(2)随着葡萄糖浓度增加,bax mRNA的表达量与Ψ丧失的间皮细胞百分率呈显著正相关(P<0.01);bcl-2 mRNA的表达量与Ψ丧失的间皮细胞百分率呈显著负相关(P<0.01),Ψ丧失的间皮细胞百分率与间皮细胞凋亡率、caspase-3活性呈显著正相关(P<0.01);(3)0.1μmol/LCsA组间皮细胞凋亡率和caspase-3活性显著低于高糖对照组(P<0.05)。结论:(1)高糖可能通过上调bax基因表达和下调bcl-2基因表达,诱导人腹膜间皮细胞线粒体活化、caspase-3活化和凋亡。(2)高糖诱导人腹膜间皮细胞caspase-3活化和凋亡可能依赖于线粒体活化。

Objective： To investigate mechanism of high-glucose-inducing injury to human peritoneal mesothelial cells. Methods： （1）The mesothelial cells were exposed to culture medium containing different concentrations of glucose （1.5%,2.5%,4.25%） for 48 hourso Then mitochondrial membrane potential （Ψ） , the apoptotic rate, caspase-3 activity, the level of bax mRNA and bcl-2 mRNA expression were measured. （2） The mesothelial cells were exposed to 4.25% glucose culture medium containing different concentrations of cyclosporin A （0.01, 0.1μmol/L） for 24 hours. Then the apoptotic rate and caspase-3 activity were measured. Results： （1） The level of bax mRNA expression of mesothelial cells, the percentage of mesothelial cells with loss of Ψ, caspase-3 activity of mesothelial cells, the apoptotic rate of mesothelial cells rose as glucose concentration increased, but the level of bcl-2 mRNA expression was decreased as glucose concentration increased （P〈0.01）. （2） There was significantly positive correlation among the level of bax mRNA expression, the percentage of mesothelial cells with loss of Ψ, caspase-3 activity and the apoptotic rate . （3） Apoptotic rate and caspase-3 activity was significantly lower in 0.1μmol/L CsA group than that in control. Conclusion： （1） High-glucose could induce loss of Ψ, apoptosis and caspase-3 activation of human peritoneal mesothelial cells by up-regulatlng expression of bax and down-regulating expression of bcl-2. （2）High-glucose could induce Caspase-3 activation in progression of high-glucose-inducing apoptosis of human peritoneal mesothelial cells by mitochondria activation.