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目的探讨沙眼衣原体质粒蛋白Pgp3诱导HeLa细胞产生IL-8的机制。方法选择不同质量浓度的Pgp3蛋白刺激HeLa细胞并在不同时间点收集细胞,Real-time PCR检测Pgp3刺激HeLa细胞后IL-8转录水平,Western blot和Real-time PCR分别检测NOD1的表达水平及下游蛋白RIP2的转录水平;Western blot检测ERK1/2和p38磷酸化水平;HeLa细胞用NOD1抑制剂ML130、RIP2抑制剂GSK583、ERK抑制剂PD98059和p38抑制剂LY2228820分别预处理后,再用Pgp3蛋白刺激细胞,ELISA和Real-time PCR检测IL-8的表达水平。结果 0~24μg/ml的Pgp3蛋白刺激HeLa细胞后,IL-8的含量呈时间和质量浓度依赖性,Pgp3蛋白刺激HeLa细胞能调控NOD1及RIP2的表达,同时ERK1/2和p38磷酸化水平显著升高;NOD1和RIP2抑制剂并不影响Pgp3蛋白诱导的ERK和p38磷酸化水平;用4种抑制剂预处理HeLa细胞后,Pgp3刺激HeLa细胞,IL-8表达量均出现下降。结论 Pgp3经NOD1/RIP2以及活化ERK1/2和p38信号通路诱导HeLa细胞调控IL-8的产生。
Objective To investigate the mechanism of Pgp3-induced IL-8 production by HeLa cells. Methods Pgp3 protein of different concentrations was used to stimulate HeLa cells and to collect cells at different time points. Real-time PCR was used to detect the transcription level of IL-8 after Pgp3 stimulated HeLa cells. Western blot and Real-time PCR were used to detect the expression of NOD1 and its downstream The protein level of RIP2 was detected by Western blot. The phosphorylation levels of ERK1 / 2 and p38 were detected by Western blot. HeLa cells were pretreated with NO1 inhibitor ML130, RIP2 inhibitor GSK583, ERK inhibitor PD98059 and p38 inhibitor LY2228820 respectively. Cell, ELISA and Real-time PCR were used to detect the expression of IL-8. Results Pgp3 protein of 0 ~ 24μg / ml stimulated HeLa cells in a time and mass concentration-dependent manner. Pgp3 stimulated HeLa cells to regulate the expression of NOD1 and RIP2, while the phosphorylation levels of ERK1 / 2 and p38 were significantly increased . NOD1 and RIP2 inhibitors did not affect Pgp3-induced phosphorylation of ERK and p38. After HeLa cells were pretreated with 4 inhibitors, the expression of IL-8 in Pgp3-stimulated HeLa cells decreased. Conclusion Pgp3 can induce HeLa cells to regulate IL-8 production through NOD1 / RIP2 and activation of ERK1 / 2 and p38 signaling pathways.