嘌呤霉素氨基核苷诱导的足细胞凋亡与COX-2表达及凋亡机制初步探讨

Expression of Cyclooxygenase-2 in Apoptotic Podocytes Induced by Puromycin Aminonucleoside

目的研究嘌呤霉素氨基核苷(PA)诱导的足细胞凋亡及环氧合酶-2(COX-2)在足细胞中的表达情况,初步探讨足细胞凋亡的机制。方法条件永生性小鼠的足细胞,经诱导分化成熟后,采用PA诱导足细胞凋亡,分别在诱导后0、8、24和48h检测足细胞活性和凋亡水平,并检测足细胞凋亡过程中caspase-3的活性水平,同时运用Westernblot技术检测足细胞P53及COX-2的表达。结果随时间的延长,PA诱导的足细胞凋亡逐渐增多,在24h及48h较明显(P<0.05)。各组细胞caspase-3水平无明显差异。24h及48hPA组P53表达较正常对照组明显增多(P<0.05)。正常足细胞表达COX-2,用PA干预后,24h及48hCOX-2表达明显增加(P<0.05)。结论PA诱导的足细胞凋亡呈时间依赖性,随时间的延长,足细胞凋亡逐渐增多;PA诱导的足细胞凋亡过程中COX-2、P53表达明显增多;足细胞凋亡过程与caspase-3无关。

Objective To investigate the expression of cyclooxygenase-2 in apoptotic podocytes induced by puromycin aminonucleoside and to study the effect of cyclooxygenase-2 on podocytes apoptosis.Methods The proliferation activity and apoptosis level of podocytes were tested at 0、8、24 and 48 h with or without puromycin aminonucleoside.Caspase-3 activity in apoptotic podocytes was tested by Caspase-3 Colorimetric Assay Kit.The p53 and cyclooxygenase-2 activity in apoptotic podocytes was tested by Western blot analysis.Results The apoptosis of podocytes induced by puromycin aminonucleoside increases over time.The significance level was found at 24 hours and 48 hours.The caspase-3 activity had no significant change in every time point.The expression of p53 in podocytes could not be invesgated,but it expressed significantly after incubated with PA for 24 hours and 48 hours.The cyclooxygenase-2 was detectable in podocytes and it was expressed much more after incubated with PA.Conclusion PA induces podocytes apoptosis in a time-dependent manner.Podocytes do not express P53.P53 expression increases in apoptotic podocytes induced by puromycin aminonucleoside.Podocytes express cyclooxygenase-2,and PA enhances the expression.Podocytes apoptosis induced by puromycin aminonucleoside is caspase-3 independent.