西安交通大学学报（医学版）2012，Vol.33Issue(1)：29-34,47,7.
沉默血小板反应蛋白-1基因对亚溶解型C5b-9诱导肾小球系膜细胞分泌细胞外基质的影响
Effects of silencing TSP-1 gene on sublytic C5b-9-induced ECM secretion from glomerular mesangial cell
摘要
Abstract
Objective To explore the effects of silencing thrombospondin-1 (TSP-1) gene on sublytic C5b-9-induced extracellular matrix (ECM) secretion from rat glomerular mesangial cell (GMC). Methods Four pairs of small hair RNA (shRNA) were designed against different regions of TSP-1 gene and the corresponding expression plasmids were constructed; the most optimized TSP-1 shRNA (shTSP-1) was then chosen to silence TSP-1 gene in the following experiment. Cultured rat GMC was transfected with shTSP-1 and stimulated with sublytic C5b-9 subsequently. We assessed the levels of TSP-1, FN and collagen IV protein expressions in GMC as well as the contents of total and active transforming growth factor-Pl(TGF-Pl) in the cultured supernatant of GMC by Western blot and ELISA, respectively. Results Nucleotide sequencing verified that the four kinds of shRNA expression vectors were all constructed successfully. Western blot assay showed that shTSP-1-3 (Thbsl-3) could effectively silence the target gene. Transfection of shTSP-1-3 into rat GMC could not only significantly decrease the expression of TSP-1 gene but also reduce sublytic C5b-9-induced activation of TGF-Pl as well as sublytic C5b-9-induced production of FN and collagen IV induced by sublytic C5b-9. Conclusion TSP-1 gene expression plays an important role in promoting TGF-Pl activation and ECM secretion in rat GMC.关键词
肾小球系膜细胞/亚溶解型C5b-9复合物/血小板反应蛋白-1/细胞外基质/小发夹RNAKey words
glomerular mesangial cell/ sublytic C5b-9/ thrombospondin-1/ extracellular matrix/ small hair RNA分类
医药卫生引用本文复制引用
邱文,李妍,周建博,赵聃,王迎伟..沉默血小板反应蛋白-1基因对亚溶解型C5b-9诱导肾小球系膜细胞分泌细胞外基质的影响[J].西安交通大学学报（医学版）,2012,33(1):29-34,47,7.基金项目
国家自然科学基金资助项目(No.31000396,81072402) （No.31000396,81072402）
江苏省高校自然科学研究项目(No.10KJB310006) （No.10KJB310006）
南京医科大学科技发展基金面上项目(No.09NJMUM003) （No.09NJMUM003）
南京医科大学基础医学院青年教师培养基金项目(No.09JC007) （No.09JC007）
