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目的探讨凝血FⅦa、Ⅹ、Ⅱa在蛋白酶激活受体2(PAR2)活化及诱导SW620细胞IL-8表达中的作用。方法用PAR2激动剂(PAR2-AP)、FⅦa、Ⅹ、Ⅱa等不同刺激物处理SW620细胞,以ELISA法检测细胞上清IL-8水平,以实时定量PCR检测细胞IL-8mRNA表达。结果血浆浓度的Ⅶa需在FⅩ存在下才能促进细胞IL-8表达;单克隆抗TF及抗PAR2抗体均可抑制FⅦa的作用;FⅡa轻微下调细胞IL-8表达,Hirudin、抗PAR1及抗PAR2抗体均可阻断此作用。结论TF-FⅦa及TF-FⅦa-FⅩa复合物,而非FⅡa,活化PAR2上调SW620细胞IL-8表达,从而促进细胞增殖及迁移。
Objective To investigate the role of coagulation FⅦa, Ⅹ and Ⅱa in the activation of PAR2 and the induction of IL-8 in SW620 cells. Methods SW620 cells were treated with PAR2 agonist (PAR2-AP), FⅦa, Ⅹ, Ⅱa and other stimuli. The level of IL-8 in supernatant of the cells was detected by ELISA. The expression of IL-8 mRNA was detected by real-time quantitative PCR. Results The plasma concentration of Ⅶ a could promote the expression of IL-8 in the presence of F Ⅹ. Both monoclonal anti-TF and anti-PAR2 antibodies could inhibit the effect of FⅦa. FⅡa down-regulated the expression of IL-8. The expression of Hirudin, anti-PAR1 and anti- Can block this effect. Conclusions TF-FⅦa and TF-FⅦa-FⅩa complex, but not FⅡa, activated PAR2 up-regulated the expression of IL-8 in SW620 cells and promoted cell proliferation and migration.