PAR_S调控大鼠HSC合成和分泌胶原蛋白的作用  

Proteinase-activated receptors regulated the expression of type I/Ⅲ collagen protein of hepatic stellate cell isolated and cultured from SD rats liver

作　　者：顾小红[1] 张云东[2] 刘迎春[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所VIP健康中心,重庆400042 [2]第三军医大学大坪医院野战外科研究所神外科,重庆400042

出　　处：《重庆医科大学学报》2010年第7期985-988,共4页Journal of Chongqing Medical University

基　　金：中国人民解放军第三军医大学中青年基金资助项目(编号:XG200543)

摘　　要：目的:探讨蛋白酶活化受体(Proteinase-activated receptors,PARs)对大鼠肝星状细胞(Hepatic stellate cell,HSC)合成和分泌胶原蛋白的调节作用。方法:体外分离和培养SD大鼠HSC,采用免疫细胞化学和RT-PCR检测PAR1、PAR2、Ⅰ和Ⅲ胶原蛋白的表达及其关系。收集细胞培养上清,并用PARs活性肽和反活性肽刺激培养细胞,检测并观察透明质酸(Hexadecanoi cacid,HA)和层粘连蛋白(Laminin,LA)的水平变化。结果:随着HSC培养时间的延长,培养细胞PAR1、PAR2表达与Ⅰ、Ⅲ胶原蛋白的表达逐渐增多,并呈一致性,14d达高峰。培养上清HA和LN水平也逐渐增高,PARs活性肽刺激培养细胞后,HA和LN表达明显增高。结论:PAR1、PAR2表达与Ⅰ/Ⅲ型胶原蛋白的表达及HA和LA的合成和分泌具有相关性,PARs对HSC的活化、合成和分泌胶原蛋白及肝纤维化的形成可能有促进作用。Objective：To investigate the role of proteinase-activated receptor 1 and 2（PAR1,PARs）in regulation ofthe expression oftype I/Ⅲ collagen protein of hepatic stellate cell isolated and cultured from SD rats liver.Methods：Hepatic stellate cell isolated and cultured from SD rats livers with a modification technique of Friedman and centrifugation of Nycodenz Gradient.Expression of mRNA for PAR1, PAR2,Ⅰcollagen and Ⅲ collagen were examined by reverse transcription polymerase chain reaction（RT-PCR）and protein examined by immunocytochemistry,observated characteristic cell shape,and examined expression of hexadecanoic acid（HA）and laminin（LN）of supernatant from culturesd HSC.Results：In the course of culturing rat HSC,vitamin A autofluorescence and lipid droplet were decreased gradually or lost following HSC activation,the cells morphology changed to stellate cells and cells proliferation activated.Ⅰ/Ⅲ collagen protein expressions were also increased,the expression of HA,LN,PAR1 and PARs paralleled with the expression ofⅠ/Ⅲ collagen protein and correlated with activation and proliferation of HSC.Conclusion：The expression of PARs correlated with activation and proliferation of HSC,could promote the expression ofⅠ/Ⅲ collagen protein,HA and LN.PARs might play an important role in the mechanisms of hepatic fibrosis.

关 键 词：蛋白酶活化受体 肝星状细胞 胶原蛋白 肝纤维化 

分 类 号：R575[医药卫生—消化系统]