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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 investigate the expression and correlation of h1 and TGF-β1 in rat mesangial cells (MsC). Methods Rat anti-thy-1 mesangial proliferative glomerulonephritis model (ATG) was prepared. The expressions of calponin h1 and TGF-β1 were detected by immunohistochemistry (ABC), Western blotting and RT-PCR respectively on the 1st, 3rd, 5th, 7th, 14th, 21st and 28th day after nephritis : The expression of calponin h1 in rat MsC was detected by immunocytochemistry and immunofluorescence assay. The expression of calponin h1 and TGF-β1 in MsC of arginine and ethanol were detected by Western blot and RT-PCR Change of expression Results The expression of calponin h1 in nephritic rats from the third day to the 14th day was significantly higher than that of the normal control group, which was little expressed in the calponin h1 and did not express TGF-β1. The expression of TGF-β1 was significantly increased in the seventh day and the 28th day (P <0.05). Calponin h1 expression localized in the cytoplasm of MsC. The expression of MsC TGF-β1 and the expression of calponin h1 were decreased after arginine treatment. The expression of calponin h1 was increased and the expression of TGF-β1 was decreased after ethanol administration. Conclusion The expression of calponin h1 in MsC of ATG rats was increased in the early stage of glomerular lesions, and later in the glomerular lesions. However, the expression of TGF-β1 was significantly increased in the middle and late glomerular lesions. calponin h1 may have negative feedback to regulate the expression of TGF-β1.