雌激素对白念珠菌诱导的小鼠腹腔巨噬细胞NLRP3表达的影响

Effect of estrogen on candida albicans-induced NLRP3 expression in murine peritoneal macrophages

目的:确定雌激素对白念珠菌诱导的小鼠腹腔巨噬细胞NLRP3炎性体基因表达的影响。方法:体外培养小鼠腹腔巨噬细胞,将其分为5组:空白对照组、白念珠菌组、白念珠菌+10^(-10)mol/L17β-雌二醇组、白念珠菌+10^(-9)mol/L 17β-雌二醇组和白念珠菌+10^(-8)mol/L17β-雌二醇组。荧光定量PCR检测细胞内NLRP3 mRNA表达;Western Blot检测细胞内NF-κBp65和caspase-1p20的活性;酶联免疫吸附法(ELISA)测定各组细胞培养上清液中白细胞介素-1β(IL-1β)的含量。结果:巨噬细胞在白念珠菌的刺激下,与空白对照组比较细胞内NLRP3 mRNA的表达、NF-κB和caspase-1p20活性和细胞培养上清液中的IL-1β含量均明显升高(P<0.01);加入雌激素干预后,随着雌激素浓度的增高,细胞内NLRP3 mRNA表达、NF-κB和caspase-1p20活性、细胞培养上清液中的IL-1β含量明显低于对照组(P<0.01)。结论:雌激素可能通过抑制NF-κB活性抑制白念珠菌诱导的NLRP3表达、caspase-1的活化和IL-1β的分泌。

Objective：To investigate the regulatory effects of estrogen on NLRP3 expression in murine per-itoneal macrophages induced by candida albicans. Methods： Murine peritoneal macrophages were cultured and randomly divided into five groups：blank control group, C. albicans group, C. albicans＋10^-10 mol/L-17β-estradiol group, C. albicans＋10^-9 mol/L- 17β-estradiol group and C. albicans＋10^-8 mol/L-17β-estradiol group.The mRNA expressions of NLRP3 in the cells were determined by real-time PCR. Western blot was per-formed to determine the activities of NF-κB and caspase-1p20. The protein of interleukin-1β（ IL-1β） were determined by enzyme-linked immunosorbent assay （ ELISA） . Results： The mRNA expressions of NLRP3, the activities of NF-κB and caspase-1p20 in macrophages and the levels of IL-1β in the supernatant in-creased significantly when compared to the controls （ P〈0.01） . After adding estrogen into the cells challenged with C. albicans, the mRNA expressions of NLRP3, the activities of NF-κB and caspase-1p20 in macropha-ges and the levels of IL-1βin the supernatant decreased with the increasing of estrogen concentrations, when compared to the control group （P〈0.01）. Conclusion： Estrogen can inhibit the expressions of NLRP3, the activation of caspase-1 and the production of pro-inflammatory cytokines IL-1β in macrophages challenged with C. albicans, through inhibiting the activities of NF-κB.