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目的研究不同免疫抑制剂对大鼠血管平滑肌细胞(VSMC)转化生长因子(TGF)-β1和Smads信号通路的影响,探讨其在慢性移植肾肾病发生、发展中的作用。方法应用大鼠胸主动脉平滑肌细胞体外原代培养技术,分别用CsA(3mg/L)、FK506(1 mg/L)、MMF(0．3mg/L)与RPM (10 mg/L)处理6、12h。用免疫组织化学和实时荧光定量聚合酶链反应检测TGF-β1、Smads蛋白和mRNA在平滑肌细胞中的表达及定位。结果CsA组和FK506组TGF-β1、Smad2的蛋白和mRNA表达水平高于对照组、MMF组和RPM组(P＜0．01),Smad7蛋白和mRNA表达低于对照组、MMF组和RPM组(P＜0．01)；MMF组和RAPA组TGF-β1、Smad2蛋白和mRNA表达低于对照组(P＜0．01),而Smad7的表达高于对照组(P＜0．01)。CsA组和FK506组,以及MMF组和RPM组的组间差异无统计学意义(P＞0．05)。结论不同免疫抑制剂可能通过影响血管平滑肌细胞TGF-β1和Smads信号通路,导致移植物动脉硬化,从而影响慢性移植肾功能丧失的进程。
Objective To investigate the effects of different immunosuppressive agents on the expression of TGF-β1 and Smads in rat vascular smooth muscle cells (VSMCs) and to explore their role in the pathogenesis and progression of chronic allograft nephropathy. Methods Primary cultured rat thoracic aorta smooth muscle cells were cultured in vitro and treated with CsA (3 mg / L), FK506 (1 mg / L), MMF (0.3 mg / L) and RPM , 12h. The expression and localization of TGF-β1, Smads protein and mRNA in smooth muscle cells were detected by immunohistochemistry and real-time fluorescence quantitative polymerase chain reaction. Results The protein and mRNA expressions of TGF-β1 and Smad2 in CsA group and FK506 group were significantly higher than those in control group, MMF group and RPM group (P <0.01), and the expressions of Smad7 protein and mRNA in CsA group and FK506 group were lower than those in control group and MMF group (P <0.01). The protein and mRNA expressions of TGF-β1 and Smad2 in MMF group and RAPA group were lower than those in control group (P <0.01), while the expression of Smad7 in MMF group and RAPA group was higher than that in control group (P <0.01). There was no significant difference between CsA group and FK506 group, MMF group and RPM group (P> 0.05). Conclusion Different immunosuppressive agents may affect arteriosclerosis of grafts and affect the process of chronic renal allograft loss by affecting the signaling pathways of TGF-β1 and Smads in vascular smooth muscle cells.