蛋白酶激活受体2激动剂诱导上皮细胞分泌MCP-1

目的:探讨蛋白酶激活受体(PAR)-2激动剂对人上皮细胞单核细胞趋化蛋白-1(MCP-1)分泌的影响.方法:人肺上皮细胞系A549细胞分别接种于12孔培养板各孔内,并分别用不同浓度的PAR-2激动剂,反PAR-2激动剂进行刺激.刺激时间为2,8和16 h.用ELISA方法检测上清液中的MCP-1水平.结果:经过16 h的培养,PAR-2激动剂SLIGKV和tc-LIGRLO均可引起浓度相关性MCP-1释放增加,tc-LIGRLO引起的最大MCP-1释放量达基础分泌量的13倍.反PAR-2激动剂对MCP-1释放的影响较小.时间相关曲线表明,PAR-2激动剂的作用从2 h开始,16 h达高峰.结论:PAR-2激动剂是高效的人肺上皮细胞MCP-1的促分泌剂.其拮抗剂有可能具有抑制呼吸道炎症的作用.

Induction of secretion of MCP-1 from epithelial cells by proteinase activated receptors-2 agonists

AIM: To investigate the actions of agonists of proteinase activated receptors (PAR)-2 on the secretion of monocyte chemoattractant protein-1 (MCP-1) from human lung epithelial cells. METHODS: A549 cells were cultured in a 12-well culture plate. The challenge was performed by adding various concentrations of PAR-2 agonists or their reverse peptides into each well, respectively. After 2 h, 8 h or 16 h, the reactions were terminated by removal of the supernatant from each well. A Sandwich ELISA was used to determine the levels of MCP-1 in supernatants. RESULTS: Following 16 h incubation, PAR-2 agonists Ser-Leu-Ile-Gly-Lys-Val-amide (SLIGKV) and trans-cinnamoyl-Leu-Ile-Gly-Arg-Leu-Orn-amide (tc-LIGRLO) were able to induce concentration dependent secretion of MCP-1. The maximum release of MCP-1 was 13 fold more than the baseline release. The reverse PAR-2 agonists had little effects on MCP-1 release. The time course showed that the actions of PAR-2 agonists initiated at 2 h and reached their peaks at 16 h. CONCLUSION: Agonists of PAR-2 are potent secretogogues of MCP-1 release from cultured human lung epithelial cells. Their antagonists may possess the ability to inhibit airway inflammation.