蛋白酶活化受体1诱导大鼠肝星状细胞分泌MCP-1

Protease-activated receptor 1 challenge rat hepatic stellate cell to produce MCP-1

目的探讨蛋白酶活化受体1(proteinase-activated receptors 1,PAR1)激动剂和活性肽对大鼠肝星状细胞(hepatic stellatecell,HSC)分泌单核细胞趋化蛋白-1(monocyte chemoattractant protein 1,MCP-1)的调控作用。方法应用改良的Friedman方法和Nycodenz一步密度梯度离心法分离SD大鼠HSC,选用生长7天的HSC,接种于12孔培养板各孔内,分别用不同浓度的PAR1激动剂-凝血酶、凝血酶抑制剂水蛭素、PAR1活性肽SFLLR和反活性肽RLLFS进行刺激,刺激时间为2、8、16 h,用ELISA方法检测培养细胞上清液中的MCP-1水平。结果经过16 h的培养,PAR1激动剂-凝血酶和活性肽SFLLR可引起HSC MCP-1的释放增加,凝血酶抑制剂水蛭素及PAR1反活性肽RLLFS不能引起MCP-1的释放增加。结论PAR1活性肽和凝血酶可促进SD大鼠HSC分泌MCP-1及肝纤维化大鼠肝脏炎症反应,PAR1反活性肽和凝血酶抑制剂在肝纤维化炎症反应中可能具有抗炎作用。

of Surgery Research, Daping Hospital, Third Military Objective To investigate the regulation effect of PAR1 agonists and active peptide on the secretion of monocyte chemoattractant protein 1 （ MCP-1 ） from rat hepatic stellate cells （HSC）. Methods Hepatic stellate cells were isolated and from SD rats livers with a modification technique of Friedmen and centrifugation of Nycodenz gradient. HSC cultured for 7 days were transferred to a 12-well culture plate. The challenge was performed by addition of various concentrations of PAR1 agonist peptides SFLLR and its reverse peptides RLLFS, thrombin or thrombin inhibitor into each well respectively. After 2 h, 8 h and 16 h, the reactions were terminated by removal of the supernatant from each well. ELISA assay was used to determine the levels of MCP-1 in supernatants. Results After 16 h incubation, SFLLR and thrombin could induce secretion of MCP-1, but the reverse PAR1 agonists-RLLFS and thrombin inhibitor-hirudin had little effects on MCP-1 release. The time course showed that the actions of PAR1 agonist peptides SFLLR and thrombin initiated at 2 h and reached their peak at 16 h. Conclusion PAR1 agonist peptides and thrombin can promote the secretion of MCP-1 from cultured rat HSC, and PAR1 antagonists and thrombin inhibitor may possess the ability to inhibit inflammation.