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目的探讨醛固酮(ALD)能否诱导培养大鼠足细胞凋亡及Akt信号通路是否参与ALD诱导的足细胞凋亡。方法体外培养并鉴定大鼠足细胞。无血清同步化处理24 h,分别给予终浓度为0(对照组)、10-9-10-5 mol／L ALD,伴或不伴螺内酯(10-7 mol／L)共孵育。流式细胞仪检测凋亡指数,Hoechst-33342染色检测足细胞凋亡。多聚酶链反应(RT-PCR)检测足细胞盐皮质激素受体(MR)和11β-羟类固醇脱氢酶2(11β-HSD2)mRNA表达。hkt活性检测试剂盒检测Akt活性表达。结果ALD以时间和剂量依赖方式诱导足细胞凋亡。螺内酯可明显抑制ALD诱导的足细胞凋亡[ALD+螺内酯组(5．61％±0．72％)比ALD组(15．10％±1．46％),P<0．05]。RT- PER结果证实足细胞表达MR和11β-HSD2 mRNA。螺内酯可部分阻断ALD对Akt活性的抑制作用。ALD诱导的足细胞凋亡率与Akt活性呈明显负相关(r=-0．77,P<0．05)。结论ALD可诱导大鼠足细胞凋亡,Akt活性下降可能参与ALD诱导的足细胞凋亡。
Objective To investigate whether aldosterone (ALD) can induce the apoptosis of podocytes and whether Akt signaling pathway is involved in ALD-induced podocyte apoptosis. Methods Rat podocytes were cultured and identified in vitro. Serum-free synchronized treatment of 24 h, respectively, to the final concentration of 0 (control group), 10-9-10-5 mol / L ALD, with or without spironolactone (10-7 mol / L) co-incubation. Apoptosis index was detected by flow cytometry and podocyte apoptosis was detected by Hoechst-33342 staining. The podocyte mineralocorticoid receptor (MR) and 11β-hydroxysteroid dehydrogenase 2 (11β-HSD2) mRNA expression were detected by polymerase chain reaction (RT-PCR) Akt activity was detected by hkt activity assay kit. Results ALD induced podocyte apoptosis in a time-and dose-dependent manner. Spironolactone significantly inhibited ALD-induced podocyte apoptosis [ALD + spironolactone (5.61% ± 0.72% vs. ALD (15.10% ± 1.46%, P <0.05]. RT-PER results confirmed podocyte expression of MR and 11 [beta] -HSD2 mRNA. Spironolactone partially blocked the inhibitory effect of ALD on Akt activity. ALD-induced rate of podocyte apoptosis was negatively correlated with Akt activity (r = -0.77, P <0.05). Conclusions ALD can induce the apoptosis of podocyte in rats and the decrease of Akt activity may be involved in ALD-induced podocyte apoptosis.