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目的研究醛固酮(ALD)及其受体拮抗剂螺内酯(SPI)对大鼠肾小球系膜细胞合成分泌纤维连接蛋白(FN),以及对转化生长因子β1(TGF-β1)基因表达的影响。方法(1)以不同浓度的ALD(10-11、10-9、10-7mol/L)及/或10-7mol/LSPI刺激肾小球系膜细胞48h后,用ELISA法测定培养上清中FN的浓度。用半定量RT-PCR法检测该细胞中FNmRNA和TGF-β1mRNA的表达。(2)以10-9mol/L的ALD刺激系膜细胞不同时间(24、48、72h)后,分别用上述方法测定培养上清中FN的浓度和细胞中FNmRNA和TGF-β1mRNA的表达。结果(1)ELISA的结果显示,ALD可促进系膜细胞合成分泌FN,且呈剂量和时间依赖性,SPI能拮抗ALD的作用。(2)半定量RT-PCR的结果显示,ALD可促进系膜细胞FNmRNA和TGF-β1mRNA的表达,也呈剂量和时间依赖性,SPI也能拮抗ALD的作用。结论ALD在蛋白和基因水平上均可促进大鼠肾小球系膜细胞合成分泌FN,及TGF-β1mRNA的表达,SPI能拮抗ALD的作用,具有一定的肾脏保护作用,从而有益于延缓肾小球硬化的进展。
Objective To investigate the effects of aldosterone (ALD) and its receptor antagonist spironolactone (SPI) on the synthesis and secretion of fibronectin (FN) and the expression of transforming growth factor β1 (TGF-β1) in rat mesangial cells. Methods (1) The mesangial cells were stimulated with different concentrations of ALD (10-11, 10-9, 10-7mol / L) and / or 10-7mol / FN concentration. Semi-quantitative RT-PCR was used to detect the expression of FN mRNA and TGF-β1 mRNA. (2) After the mesangial cells were stimulated with 10-9mol / L ALD for different time (24,48,72h), the concentration of FN and the expression of FN mRNA and TGF-β1 mRNA in the culture supernatant were measured by the above methods respectively. Results (1) The results of ELISA showed that ALD could promote the synthesis and secretion of FN by mesangial cells in a dose and time dependent manner. SPI could antagonize the effect of ALD. (2) The results of semi-quantitative RT-PCR showed that ALD could promote the expression of FN mRNA and TGF-β1 mRNA in mesangial cells in a dose and time dependent manner, and SPI could also antagonize the effect of ALD. Conclusion ALD can promote the synthesis and secretion of FN and the expression of TGF-β1 mRNA in rat mesangial cells both at the protein and gene level. SPI can antagonize the effect of ALD and have a protective effect on kidney, Progress in the hardening of the ball.