螺内酯对高糖及醛固酮诱导的肾小球系膜细胞CTGF表达的影响

Effects of spironolactone on aldosterone or high glucose induced CTGF mRNA and protein expression in the glomerular mesangial cells

目的通过研究螺内酯(SPI)对高糖(HG)和醛固酮(ALD)刺激的体外培养的大鼠肾小球系膜细胞(MsC)结缔组织生长因子(CTGF)mRNA和蛋白表达的影响,探讨SPI的肾脏保护机制。方法将大鼠MsC分为:A组,正常对照组(5.6 mmol.L-1)、B组,高糖组(30 mmol.L-1)、C组,HG+SPI干预组(10-9,10-8,10-7mol.L-1)、D组,ALD组(5.6 mmol.L-1葡萄糖+10-7mol.L-1ALD)、E组,ALD+SPI干预组(10-9,10-8,10-7mol.L-1)。酶联免疫法检测上清液中CTGF蛋白的含量,RT-RCR法检测MsC CTGFmRNA表达。结果①正常糖浓度的培养基中可测到MsCCTGF mRNA和蛋白表达;②与A组比较,B组和D组MsCCTGF mRNA表达和上清液CTGF水平明显升高,差异有显著性;③与B组比较,C组中经10-8,10-7mol.L-1SPI干预MsC CTGF mRNA表达和上清液CTGF水平明显降低,P<0.05,但10-9mol.L-1SPI干预MsC CTGF mRNA表达和上清液CTGF水平无变化,P>0.05;④与D组比较,E组MsCCTGFmRNA表达和上清液CTGF水平明显降低,P<0.05。结论 SPI可抑制HG和ALD刺激的大鼠MsC CTGFmRNA表达和蛋白合成,该作用可能是其抗纤维化和防治肾损害的机制之一。

Aim To study the renoprotective mechnisms of spironolactone（SPI）by observing its effect on the expression of connective tissue growth factor（CTGF）mRNA and protein in rat mesangial cells（MsC）cultured in aldosterone（ALD）or high glucose（HG）induced state.Methods MsC were divided into the following groups：group A,the normal glucose group（NG,5.6 mmol·L-1 Glu）,group B,high glucose group,group C,HG＋SPI（10-9,10-8,10-7 mol·L-1）group,group D,ALD groups（NG＋10-7 mol·L-1 ALD）,group E,ALD＋SPI（10-9,10-8,10-7 mol·L-1）group.The expressions of CTGF mRNA in MsC were detected by semi-quantitative RT-PCR,while the levels of CTGF protein in supernatants were measured by ELISA.Results ① MsC expressed CTGF mRNA and protein in normal glucose condition;② The CTGF mRNA in the MsC and CTGF protein values in the supernatant in group B and group D increased significantly compared to group A（P〈0.05）.ALD induced CTGF mRNA and protein expressions of MsC markedly down-regulated in 10-9,10-8 and 10-7 mol·L-1 SPI-treated group.However,in high glucose condition there was no statistical difference in the levels of CTGF mRNA and protein between group 10-9 mol·L-1 SPI-treated group and group B（P〉0.05）.Conclusion Spironolactone can down-regulate the CTGF mRNA and protein expression induced by ALD and HG in glomerlar MsC,which may provide an experimental evidence for further evaluating the possible protective effect of SPI against diabetic nephropathy.