摘要
目的 通过对大鼠肾小球系膜细胞(mesangial cell,MsC)转染Smad7基因,观察转染阳性细胞克隆uPA及PAI-1表达的改变,以进一步阐明Smad7阻断组织纤维化过程的作用机制。方法 经脂质体介导将含有Smad7重组表达质粒转染大鼠MsC,用G418筛选及RT-PCR、Western blot法鉴定;又分别采用RT-PCR与Western blot法,检测转染阳性细胞克隆uPA及PAI-1表达改变。结果 成功建立高表达Smad7的阳性MsC克隆(S-22、S-26),并证实其uPA mRNA及蛋白表达明显增加,而PAI-1 mRNA及其蛋白的表达显著下降。结论 Smad7可能通过增强组织内uPA酶的生成和减少PAI-1的合成而减轻组织纤维化进展。
Purpose: To elucidate the mechanism of Smad7 blocking tissue fibrosis by observing urokinase-type plasminogen activator(uPA) and plasminogen activator inhibitor-1 (PAI-1) expressions on cultured rat mesangial cells(MsC) transfected with Smad7 cDNA. Methods: Lipofectin method was used to transfect Smad7 cDNA into rat MsC, RT-PCR and Western blot analysis for detecting Smad7 mRNA and protein expression level. The expressions of uPA and PAI-1 were also determined by RT-PCR and Western blot. Results: Overexpression of Smad7 on two transfected MsC clones (S-22, S-26) were successfully established. Two MsC clones showed increased expressions of uPA its mRNA and protein, accompanied with decreased synthesis of its mRNA and PAI-1 protein. Conclusions: It is possible that Smad7 can alleviate the development of tissue fibrosis by upregulating the expressions of uPA and decreased synthesis of PAI-1.
出处
《复旦学报（医学版）》
EI
CAS
CSCD
北大核心
2004年第1期5-8,共4页
Fudan University Journal of Medical Sciences
基金
上海科技发展基金(01JC14018)
