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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 apoptosis of podocytes induced by puromycin aminonucleoside (PA) and the expression of cyclooxygenase-2 (COX-2) in podocytes, and to explore the mechanism of podocyte apoptosis. Methods The podocytes of immortalized mice were induced to differentiate into mature podocytes and then induced with pA induced apoptosis of podocytes. The podocyte viability and apoptosis were detected at 0, 8, 24 and 48 h after induction, and podocyte apoptosis The level of caspase-3 activity in podocytes and the expression of P53 and COX-2 in podocytes were detected by Western blotting. Results With the prolongation of time, the number of podocytes induced by PA increased gradually and reached the peak at 24h and 48h (P <0.05). There was no significant difference in the levels of caspase-3 in each group. P53 expression in 24h and 48hPA group was significantly higher than that in control group (P <0.05). COX-2 was expressed in normal podocytes, and COX-2 expression was significantly increased at 24h and 48h after PA intervention (P <0.05). Conclusion PA induced podocyte apoptosis in a time-dependent manner, with the passage of time, podocyte apoptosis gradually increased; PA-induced podocyte apoptosis in the process of COX-2, P53 expression was significantly increased; podocyte apoptosis and caspase -3 does not matter.