大鼠系膜细胞中碱性调宁蛋白h1和转化生长因子β1的表达及其相互关系

Relationship between calponin h1 and TGF-β1 in rat mesangial cells

目的 探讨大鼠系膜细胞(MsC)中碱性调宁蛋白(calponin)h1与TGF-β1的表达 及其相互关系。方法制备大鼠抗thy-1系膜增生性肾炎模型(ATG)。应用免疫组织化学ABC 法、Western印迹、RT-PCR等方法分别从蛋白和核酸水平检测calponin h1和TGF-β1在肾炎模型 第1、3、5、7、14、21、28天组织中的表达:采用细胞免疫化学和细胞免疫荧光法检测培养大鼠 MsC中calponin h1的表达;应用Western印迹和RT-PCR的方法观察精氨酸和乙醇分别孵育后 大鼠MsC中calponin h1和TGF-β1的表达变化。结果 与少量表达calponin h1和不表达TGF-β1 的正常组比,肾炎3 d组至14 d组calponin h1表达明显增强,7 d组至28 d组TGF-β1表达显 著增强(P<0.05)。calponin h1表达定位于大鼠MsC胞浆内。精氨酸作用后大鼠MsC TGF-β1表 达增强,calponin h1表达减弱;乙醇作用后大鼠MsC表达calponin h1升高,表达TGF-β1降低。 结论 ATG大鼠MsC中calponin h1表达在肾小球病变早期增强.后期减弱;而TGF-β1表达则 在肾小球病变中晚期显著增强。calponin h1可能具有负反馈调节TGF-β1表达的作用。

Objective To explore the relationship between calponin h1 and TGF-β1 in rat MsC.Methods Anti-thy-1 glomerulonephritis(ATG) rat model was established by injecting anti-thy-1 polyclonal antibody.Total mRNA and protein were extracted from the renal cortex of sacrificed rats 1 , 3, 5, 7, 14, 21, 28 days after injection respectively. Immunohistochemistry, Western blot, RT-PCR were used to detect the expression of calponin h1 and TGF-β1 .Cytoimmunochemistry and cytoimmunofluorescence were performed to identify the expression of calponin h1 in rat MsC. The expression of calponin h1 and TGF-β1 in rat MsC treated with exeess arginine and ethanol respectively was evaluated by Western blot and RT-PCR analysis. Results In comparison to weak calponin h1 expression and no TGF-β1 expression in control group, calponin h1 expression increased from day 3 lo day 14, whereas TGF-β1 expression greatly increased from day 7 to day 28 (P < 0.05). Calponin h1 expression was located in cytoplasm of cultured rat MsC. Rat MsC grown in excess arginine increased its TGF-β1 expression and decreased its calponin h1 expression obviously compared with other two groups. On the contrary, rat MsC incubated with excess ethanol upregulated its protein and mRNA expression of calponin h1 and downregulated its mRNA expression of TGF-β1. Conclusions Calponin h1 expression is enhanced in the early stage of ATG.With the progression of ATG, calponin h1 expression decreases accompanied by an evident increase of TGF-β1 expression. Calponin h1 may have an negative regulatory effect on TGF-β1 expression in rat MsC.