【摘要】 目的初步探讨鹦鹉热嗜衣原体蛋白CPSIT_p7对宿主细胞炎症反应的调节作用及其分子机制。方法佛波酯(PMA)处理THP-1细胞过夜,诱导其分化为贴壁的巨噬细胞,之后用CPSIT_p7蛋白刺激贴壁细胞,或先用30μmol/L ERK抑制剂PD98059、JNK抑制剂SP600125和p38抑制剂SB202190分别预处理贴壁细胞,再用CPSIT_p7蛋白处理贴壁细胞;Western blot检测ERK、JNK和p38磷酸化水平,ELISA检测各种炎症因子的表达水平。结果 0~10μg/ml CPSIT_p7蛋白刺激PMA诱导的THP-1细胞24 h后,随着CPSIT_p7质量浓度升高,IL-6、IL-1β、IL-8及TNF-α的含量呈剂量依赖性增加;10μg/ml的CPSIT_p7处理细胞0、6、12、24和36 h,在24 h时IL-6、IL-8及IL-1β表达水平达到高峰,而TNF-α在12 h就达到高峰;CPSIT_p7蛋白处理细胞后其ERK和JNK磷酸化水平显著升高,p38磷酸化水平改变不明显;JNK和ERK抑制剂能明显降低CPSIT_p7蛋白诱导的IL-6、IL-1β、IL-8及TNF-α表达。结论 CPSIT_p7通过JNK/MAPKs和ERK/MAPKs信号传导途径诱导THP-1产生IL-1β、IL-6、IL-8及TNF-α炎症因子,与p38/MAPKs信号传导通路无关。更多还原

【Abstract】 In the present study,we explored the mechanism of the CPSIT_p7 from C.psittaci in regulating the inflammatory response of host cells.THP-1 cells were incubated with CPSIT_p7,or ERK inhibitor(PD98059)/JNK inhibitor(SP600125)/p38 inhibitor(SB202190) plus CPSIT_p7 at different time points,and several inflammatory cytokines in the supernatant were tested.The phosphorylated proteins of JNK,ERK and p38 were detected by Western blotting.The results showed that CPSIT_p7 induced THP-1 cells to produce CKs in a dose-dependent manner.When THP-1 cells were incubated with 10 μg/ml of CPSIT_p7,the expression of IL-6,IL-8 and IL-1β reached the highest level at 24 h and TNF-α expression reached the highest level at 12 h.The ERK and JNK inhibitors decreased the levels of IL-8,IL-6,TNF-α and IL-1β.Furthermore,CPSIT_p7 promoted phosphorylation of ERK and JNK,not p38 in THP-1 cells.We conclude that CPSIT_p7 induces THP-1 cells to produce inflammatory cytokines such as IL-1β,IL-6,IL-8 and TNF-α by activating the JNK or ERK signaling pathways.更多还原