摘要
目的 通过观察转染DCN基因的大鼠系膜细胞 (MsC)生长及其一些表型的改变 ,为其用作细胞载体回输入大鼠肾病模型体内进行基因治疗奠定实验基础。方法 采用MTT比色法和流式细胞仪技术 ,检测该MsC株生长情况 ;Northernblot和Westernblot法检测其TGF β1mRNA、ColⅣmRNA和TGF β1蛋白表达水平 ;半定量RT PCR法检测其TIMP 2mRNA表达的改变。结果 与未转染MsC相比 ,转染DCN基因的MsC株的生长受到明显抑制 (5d时P <0 .0 5 ,6d时P <0 .0 1) ;G0 /G1期比例增加 ,S期比例降低 ,提示其生长缓慢 ;TGF β1和ColⅣmRNA表达明显降低 ,TGF β1蛋白分泌减少 ;TIMP 2mRNA表达明显下调。 结论 转染DCN基因的MsC可被用作载体开展对大鼠系膜增生性肾炎的实验治疗。
Purpose: To study the growth and the changes of some phenotypes of cultured rat mesangial cell (MsC) transfected by decorin(DCN) gene in order to provide experimental basis for gene delivery to diseased glomeruli of rat mesangial proliferative glomerulonephritis. Methods: We applied MTT assay and FCM analysis for examining MsC growth, Northern blot and Western blot analyses for measuring the levels of transforming growth factor-β1 (TGF-β1) mRNA and its protein, and Col IV mRNA expressions respectively, semi-quatification RT-PCR method for detecting the level of type-2 tissue inhibitor of metalloproteinase(TIMP-2) mRNA expression on MsC. Results: The inhibited growth, increased G0/G1 phase percentage and decreased S phase percentage of MsC clones transfected by DCN gene were observed by MTT method and FCM analysis respectively, compared to that of the untransfected MsC. The levels of TGF-β1 mRNA and Col IV mRNA expressions and TGF-β1 protein synthesis were decreased by Northern blot and Western blot analyses respectively, and TIMP-2 mRNA expression level of the transfectants were decreased by RT-PCR, compared to that of the untmsfected MsC. Conclusions: DCN gene transfected to MsC can inhibit MsC growth and ECM synthesis in varying degrees.
出处
《复旦学报（医学版）》
EI
CAS
CSCD
北大核心
2003年第5期414-417,共4页
Fudan University Journal of Medical Sciences
基金
上海市教委重点学科基金 (B990 80 2 )
教育部博士点基金 ( 2 0 3 1)
国家自然科学基金 ( 3 10 70 43 1)资助项目
