Sublytic C5b-9诱导肾小球系膜细胞产生的TSP-1和TGF-β1对其合成FN和collagenⅣ的影响  

The effects of TSP-1 and TGF-β1 induced by sublytic C5b-9 on FN and collagen Ⅳ synthesis from glomerular mesangial cell

作　　者：邱文[1] 车楠[1] 周建博[1] 李妍 赵聃[1] 王迎伟[1] 

机构地区：[1]南京医科大学微生物与免疫学系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2011年第11期1543-1549,共7页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助(31000396;81072402);江苏省高校自然科学研究(10KJB310006);南京医科大学科技发展基金面上项目(09NJMUM003);南京医科大学基础医学院青年教师培养基金(09JC007)

摘　　要：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 explore the roles of thrombospondin-1（TSP-1） and transforming growth factor-β1（TGF-β1） in mediating the secretion of extracellular matrix（ECM）,such as fibronectin（FN） and collagen Ⅳ,from rat glomerular mesangial cell（GMC） induced by sublytic C5b-9 complexes.Methods： Rat GMC was cultured and divided into different groups.The expression levels of TSP-1 and TGF-β1 as well as FN and collagen Ⅳ in GMC with sublytic C5b-9 stimulation were measured.In addition,the synthetic blocking peptide of TSP-1（GGWSHW） and TGF-β1 neutralizing antibody were utilized to explore the role of TSP-1 and TGF-β1 in ECM secretion from GMC induced by sublytic C5b-9,as well as the relationship between TSP-1 and TGF-β1.Results： The protein levels of FN and collagen Ⅳ as well as TSP-1 and TGF-β1 including the activated TGF-β1 were significantly increased in rat GMC at 18 h after sublytic C5b-9 stimulation.The blocking peptides of TSP-1（GGWSHW） could significantly inhibit TGF-β1 activation and reduce the production of FN and collagen Ⅳ.In addition,TGF-β1 neutralizing antibody could also obviously inhibit the expression of FN and collagen Ⅳ in GMC upon sublytic C5b-9 attack.Conclusion： Sublytic C5b-9 can promote ECM secretion and TSP-1 and TGF-β1 production as well as TGF-β1 activation in rat GMC in vitro,while the secretion of ECM stimulated by sublytic C5b-9 is possibly mediated in part by TSP-1-regulated TGF-β1 activation.

关 键 词：sublytic C5B-9 肾小球系膜细胞 血小板反应蛋白-1 转化生长因子-β1 纤维连接蛋白 Ⅳ型胶原蛋白 

分 类 号：R692.3[医药卫生—泌尿科学]