论文部分内容阅读
目的:探讨亚溶解型(sublytic)C5b-9复合物刺激大鼠肾小球系膜细胞(glomerular mesangial cell,GMC)后,诱生的血小板反应蛋白-1(thrombospondin-1,TSP-1)与转化生长因子-β1(transforming growth factor-β1,TGF-β1)对其促进细胞外基质(ex-tracellular matrix,ECM),如纤维连接蛋白(fibronectin,FN)和Ⅳ型胶原蛋白(collagenⅣ)合成的影响。方法:体外培养大鼠GMC,进行不同分组处理并给予sublytic C5b-9刺激,然后分别检查受刺激的GMC合成TSP-1和TGF-β1因子水平,同时检测GMC分泌FN和collagenⅣ的水平。此外,应用人工合成的TSP-1封闭肽段(GGWSHW)和TGF-β1中和抗体处理培养的大鼠GMC,研究TSP-1和TGF-β1在sublytic C5b-9诱导的GMC分泌上述ECM中的作用及其相互关系。结果:培养的大鼠GMC在sublytic C5b-9刺激后18 h,其FN和collagenⅣ表达水平均明显升高。同时,TSP-1蛋白表达和TGF-β1分泌(包括活化的TGF-β1含量)也显著增多。用TSP-1封闭肽段(GGWSHW)处理大鼠GMC后亦能显著抑制由sublytic C5b-9诱导的TGF-β1活化,并减少FN、collagenⅣ的产生。同样用TGF-β1中和抗体处理GMC后也能明显抑制由sublytic C5b-9导致的FN、collagenⅣ的分泌。结论:体外用sublytic C5b-9刺激大鼠GMC,能诱导其ECM分泌与TSP-1和TGF-β1的合成及活化,而sublytic C5b-9促进GMC分泌ECM的机制可能与其诱导TSP-1合成及活化的TGF-β1存在一定的关系。
Objective: To investigate the effect of TSP-1 induced by sublytic C5b-9 complex on rat glomerular mesangial cells (GMC) The effect of transforming growth factor-β1 (TGF-β1) on the synthesis of ex-tracellular matrix (ECM) such as fibronectin (FN) and collagen Ⅳ influences. Methods: GMCs were cultured in vitro and treated with sublytic C5b-9 in different groups. The levels of TSP-1 and TGF-β1 in stimulated GMCs and the levels of FN and collagenⅣ secreted by GMCs were measured. In addition, cultured rat GMCs were treated with synthetic TSP-1 blocking peptide (GGWSHW) and TGF-β1 neutralizing antibody to study the effect of TSP-1 and TGF-β1 in the secretion of subcellularly-derived GMCs in sublytic C5b-9 Role and their relationship. Results: The expression of FN and collagen Ⅳ in cultured GMCs of rats was significantly increased at 18 h after sublytic C5b-9 stimulation. At the same time, TSP-1 protein expression and TGF-β1 secretion (including activated TGF-β1 content) also increased significantly. Treatment of rat GMC with the TSP-1 blocking peptide (GGWSHW) also significantly inhibited TGF-β1-induced activation of sublytic C5b-9 and reduced FN and collagenⅣ production. The same treatment with TGF-β1 neutralizing antibody also significantly inhibited the secretion of FN and collagen Ⅳ by sublytic C5b-9. CONCLUSIONS: In vitro stimulation of GMC by sublytic C5b-9 induces the secretion of ECM and the synthesis and activation of TSP-1 and TGF-β1, and the mechanism by which sublytic C5b-9 promotes the secretion of ECM by GMC may be related to its induction of TSP-1 synthesis and Activated TGF-β1 there is a certain relationship.