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目的探讨体外醛固酮(ALD)对大鼠肾小管上皮细胞凋亡基因的影响。方法体外培养大鼠肾小管上皮细胞NRK-52E。台盼兰染色法检测ALD浓度为10-7、10-6、10-5、10-4和10-3mol/L对NRK-52E细胞的毒性。采用流式细胞术检测ALD浓度为10-6 mol/L(ALD组)和正常对照组细胞凋亡率;RT-PCR检测两组凋亡相关基因p53、Bcl-2和C-myc mRNA表达。结果 ALD浓度为10-7、10-6和10-5 mol/L对NRK-52E细胞均无明显毒性作用。ALD组细胞凋亡率高于正常对照组[(63.72±2.50)%vs.(51.52±4.30)%](P<0.05);与正常对照组比较,ALD组(10-6 mol/L)p53mRNA和C-myc mRNA水平升高,而Bcl-2mRNA水平下降(P<0.05)。结论 ALD可能通过上调p53、C-myc基因表达和下调Bcl-2基因表达诱导肾小管上皮细胞凋亡。
Objective To investigate the effect of aldosterone (ALD) on the apoptosis of rat renal tubular epithelial cells. Methods Rat renal tubular epithelial cells NRK-52E were cultured in vitro. Trypan blue staining was used to determine the toxicity of ALD at concentrations of 10-7, 10-6, 10-5, 10-4 and 10-3 mol / L on NRK-52E cells. The apoptotic rates of ALD group and normal control group were detected by flow cytometry. The expressions of p53, Bcl-2 and C-myc mRNA were detected by RT-PCR. Results The ALD concentration of 10-7, 10-6 and 10-5 mol / L had no obvious toxic effect on NRK-52E cells. The apoptosis rate of ALD group was significantly higher than that of normal control group [(63.72 ± 2.50)% vs (51.52 ± 4.30)%] (P <0.05). Compared with normal control group, And C-myc mRNA levels increased, while Bcl-2 mRNA levels decreased (P <0.05). Conclusion ALD may induce the apoptosis of renal tubular epithelial cells by up-regulating the expression of p53, C-myc gene and down-regulating Bcl-2 gene expression.