α-硫辛酸对高糖 高同型半胱氨酸诱导的小鼠足细胞凋亡的影响

Influence of alpha-lipoic acid on apoptosis of mouse podocytes cultured in high-level glucose and homocysteine

目的探讨α-硫辛酸对高糖、高同型半胱氨酸(Hcy)作用下小鼠足细胞凋亡及Nephrin mRNA表达的影响。方法将体外培养的足细胞用高糖(25mmol/L)、Hcy(1mmol/L)、α-硫辛酸(0.5mmol/L)干预,分别在干预24、48、72h时收集细胞,流式细胞仪检测细胞凋亡,反转录聚合酶链反应(RT-PCR)法检测Nephrin mRNA表达。结果 24h时,高糖组、高Hcy组及混合组未见明显细胞凋亡(P>0.05);48h时,上述3组可见明显的细胞凋亡,且混合组凋亡更加明显(P<0.01);72h时,上述差异更加明显(P<0.01);α-硫辛酸可对抗高糖、高Hcy引起的足细胞凋亡(均P<0.05),且有一定的时间依赖性;高糖、高Hcy环境下,足细胞Nephrin mRNA表达呈下降趋势,且有一定的时效性;α-硫辛酸可对抗上述趋势。结论α-硫辛酸可减弱高糖、高Hcy对足细胞促凋亡作用,还可对抗其下调Nephrin mRNA的表达。

Objective To observe the influences of alpha-lipoic acid on apoptosis of mouse podocytes cultured in high glucose （Hg） and high homoeysteine （Hey） levels, and on expression of Nephrin mRNA in mouse podocytes. Methods The podocytes were cultured in high-level glucose （25 retool/L）, high-level Hey （l mmol/L） and alphalipoic acid （0.5 retool/L） before they were collected respectively at 24, 48 and 72 hours. Cell apoptosis was detected by flow cytometry and mRNA expression of Nephrin was examined by RT-PCR. Results No obvious apoptosis was observed at 24 hours in the Hg, Hey and mixed groups （P〉0.05）. At 48 hours obvious cell apoptosis was observed in the Hg, Hey and mixed groups compared with the normal control group （P〈0.05） and the apoptosis rate in the mixed group was significantly higher than in other groups （P〈0.01）. At 72 hours the apoptosis in each group was significantly increased than at 48 hours （P〈0.01）. Alpha-lipoic acid had an agonistic effect in a time-dependent manner on the apoptosis of podocytes induced by Hg and Hcy （all P〈0.05）. The expression of Nephrin mRNA was decreased in mouse podocytes cultured in Hg and Hcy also in a time-dependent manner, but alpha-lipoic acid had an agonistic effect on the trend. Conclusion Alpha-lipoic acid can attenuate the effect of high glucose and high Hcy on the apoptosis of podocytes, and can also counter the down-expression of Nephrin mRNA in mouse podocytes induced by high glucose and high Hcy.