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目的:研究趋化因子RANTES对肥大细胞膜表面与细胞质内蛋白酶激活受体(protease-activated receptors,PARs)各亚型表达的影响。方法:肥大细胞瘤细胞P815以不同浓度RANTES(0.1、1.0、10.0、100.0 ng/ml)激发,于不同时间点(2、6及16h)收集细胞,予透膜及非透膜处理,流式细胞术检测全细胞与细胞膜PARs的表达水平,分析细胞膜与胞质中PARs的表达变化。结果:RANTES作用6 h可显著下调细胞膜表面PAR1,2 h及16 h有下调趋势但差异无统计学意义,该下调效应可被抗RANTES单克隆抗体(anti-RANTES)取消;透膜处理后分析结果显示,RANTES对细胞质内PAR1表达无显著调节作用,细胞膜与细胞质中PAR1表达差异无统计学意义。RANTES对P815细胞PAR2、PAR3、PAR4表达均无显著调节作用。结论:RANTES可选择性调节肥大细胞膜表面PAR1表达水平,对细胞质内PAR1及其他PARs亚型(PAR2、PAR3、PAR4)胞膜与胞质水平均无显著影响。
Objective: To investigate the effect of chemokine RANTES on the expression of various subtypes of protease-activated receptors (PARs) on the surface of mast cells and in cytoplasm. Methods: M8 cells were stimulated with different concentrations of RANTES (0.1, 1.0, 10.0 and 100.0 ng / ml). The cells were harvested at different time points (2, 6 and 16 h) The expression of PARs in whole cell and cell membrane was detected by cytometry, and the expression of PARs in cell membrane and cytoplasm was analyzed. RESULTS: After treated with RANTES for 6 h, the down-regulation of PAR1 was observed at 2 h and 16 h, but the difference was not statistically significant. The anti-RANTES effect could be abolished by anti-RANTES monoclonal antibody. The results showed that RANTES had no significant effect on the expression of PAR1 in the cytoplasm, but there was no significant difference in the expression of PAR1 between the cell membrane and cytoplasm. RANTES had no significant effect on the expression of PAR2, PAR3 and PAR4 in P815 cells. CONCLUSION: RANTES can selectively regulate the expression of PAR1 on the surface of mast cell membrane, but have no significant effect on the membrane and cytoplasm of PAR1, PAR2 and PAR4 in the cytoplasm.