雌二醇对Hela细胞淋球菌感染模型il-6、il-8、nlrp3表达的影响

Effect of estradiol on the expression of il-6, il-8 and nlrp3 in Hela cell gonococcal infection model

目的了解17β-雌二醇(E2)对人HeLa细胞淋球菌(Ng)感染模型中细胞因子il-6、il-8、nlrp3 mRNA表达的影响。方法体外培养人HeLa细胞,分为如下4组:对照组、雌二醇组(E2组)、淋球菌感染组(Ng组)、淋球菌+雌二醇组(Ng+E2组);对照组予DMEM培养基,E2组予17β-雌二醇+DMEM培养,Ng组予淋球菌悬液+DMEM培养,Ng+E2组予17β-雌二醇+淋球菌悬液+DMEM培养,培养6、18 h后应用荧光定量PCR(q R检测各组细胞il-6、il-8、nlrp3T-PCR)m RNA表达量。结果淋球菌刺激HeLa细胞6、18 h后,Ng mRNA组的il-6、il-8、nlrp3水平较对照组明显增加(P<0.01),组il-6、il-8Ng+E2 mRNA高于对照组但较Ng组下调(P<0.01或,而nlrp30.05)m RNA表达明显低于Ng组及对照组(P<0.01或0.05);E2组刺激6 h的il-8和刺激18 h的nlrp3与对照组差异无统计学意义(P>0.05);其余指标差异均有统计学意义(P<0.01或0.05)。结论 E2可下调淋球菌感染诱导的人中il-6、il-8、nlrp3HeLa mRNA等促炎因子的表达,起到抑制黏膜免疫,促进淋球菌感染的作用。

Objective To investigate the regulatory effect of E2 on the expression of inflammatory cytokine（il-6, il-8,nlrp3 Methods） mRNA in human Hela cells pretreated with Neisseria gonorrhoeae（Ng）. HeLa cells were cultured in vitro, which were randomly divided into four groups： Control Group, 17β-Estradiol Group（E2 Group）, Neisseria Gonorrhoeae Group（Ng Group）, Neisseria Gonorrhoeae ＋17β-Estradiol Group（E2＋Ng Group）. The Control Group was cultured in DMEM; The E2 Group was cultured in DMEM plus 17β-estradiol; The Ng Group was cultured in DMEM plus gonococcal suspension; The E2＋Ng Group was cultured in the same condition as the Ng Group plus 17β-Estradiol. Total RNA was isolated after 6 and 18 hours, and the expression of il-6,il-8,nlrp3 mRNA in each group was determined by q PCR. ResultsThe expression level of il-6, il-8 and nlrp3 mRNA in Ng Group was significantly increased compared with the Control Group after Neisseria gonorrhoeae stimulated Hela cells for 6 h and 18 h（P〈0.01）. The expression of il-6 and il-8 mRNA in Ng＋E2 Group was higher than that of the Control Group but down-regulated compared with the Ng Group（P〈0.01 or 0.05）, and the expression of nlrp3 mRNA of the Ng Group was significantly lower than that of the Ng Group and Control Group（P〈0.01 or 0.05）; There was no statistical difference in the expression of il-8 after 6 h stimulation and the expression of nlrp3 after 18 h stimulation between the E2 Group and the Control Group（P〉0.05）; and there have been statistical differences for all the other indicators（P〈0.01 or 0.05）. Conclusion E2 can down-regulate the expression of the il-6, il-8, nlpr3 and other inflammatory factors in human Hela induced by gonococcus and thus achieve the effect of inhibiting mucosal immunity and promoting gonococcal infection.