目的检测肿瘤坏死因子(TNF)引起的肥大细胞蛋白酶激活受体(PAR).1,2,3,4表达。方法肥大细胞P815培养后以不同浓度的TNF激发肥大细胞,在不同时间点收集细胞,给予聚乙二醇辛基苯基醚(TritonX一100)非TritonX-100处理后用流式细胞术和激光共聚焦方法检测肥大细胞蛋白酶激活受体的表达。结果TNF激发肥大细胞2h、6h和16h后,TritonX.100与非TritonX.100处理后,肥大细胞PAR-1,3表达均无明显变化;但TNF以浓度依赖方式上调P515肥大细胞PAR.2,4表达(P〈0.05);上述各时间点检测PAR-1,2,3表达情况,TritonX.100处理组与非TritonX.100处理组检测结果无明显差异,但PAR-4表达结果显示TritonX-100处理组表达强于非TritonX.100处理组。结论TNF可以上调P815肥大细胞PAR-2,4表达,但对PAR-1,3表达的调节作用不明显,TritonX-100处理和非TritonX-100处理不影响TNF调节的肥大细胞PARs表达的检测结果。
Objective To investigate the potential i-nfluence of TNF on the expression of protease activated receptor (PAR)-1,2,3 and 4 by using P815 mast ceils. Methods After being challenged with various concentrations of TNF for 2 h, 6 h and 16 h, the P815 mast cells were treated with or without Triton X-100 and the PAR expressions were detected by flow eytometry and immunofluorescent microscopy. Results Compared with the corresponding controls, TNF concentration-dependently upregnlated expressions of PAR-2 and PAR-4 both in Triton X-100-treated and the untreated groups, but had no significant effect on the ex- pression of PAR-1 and PAR-3. Moreover, no significantly different expressions of TNF-indueed PAR-l, 2, 3 were observed between Triton X-100-treated and the untreated groups, whereas Triton X-100-treated PAR-4 expressions were significantly enhanced by TNF compared with the Triton X-100-untreated ones. Conclusion TNF can up-regnlate PAR-2, 4 expression of P815 mast cells but has little effect on the expression of PAR- 1, 3 correspondingly. And Triton X-100 treatment had no significant effect on TNF-moclulated expression of PARs in P815 mast cells.