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目的 探讨孕激素对淋球菌(Ng)诱导的人宫颈癌细胞(Hela)NALP3炎性体基因表达的影响。方法 将体外培养的Hela细胞分为5组,即空白对照组、Ng组、Ng+10-9mol/L孕酮组、Ng+10-8mol/L孕酮组和Ng+10-7mol/L孕酮组。荧光定量PCR检测Hela细胞内NALP3mRNA表达,WesternBlot检测Hela细胞内NF-κBp65和caspase-1p20活性的变化,酶联免疫吸附试验法(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 Hela细胞在淋球菌的刺激下,细胞内NALP3mRNA表达、NF-κB和caspase-1p20活性、细胞培养上清液中的IL-1β含量均明显升高,Ng组与空白对照组比较差异有统计学意义(P<0.01);加入孕酮干预后,随着孕酮浓度的增高,细胞内NALP3mRNA表达、NF-κB和caspase-1p20活性、细胞培养上清液中的IL-1β含量明显降低,与Ng组比较,差异均有统计学意义(P均<0.01)。结论 孕酮可抑制淋球菌诱导的NALP3表达及caspase-1的活化和IL-1β的分泌,且这种抑制作用可能是通过抑制NF-κB活性实现的。
Objective To investigate the effects of progesterone on the gene expression of NALP3 induced by Neisseria gonorrhoeae (Ng) in human cervical carcinoma cells. Methods Hela cells cultured in vitro were divided into 5 groups: blank control group, Ng group, Ng + 10-9mol / L progesterone group, Ng + 10-8mol / L progesterone group and Ng + 10-7mol / L Ketone group. The expression of NALP3mRNA in Hela cells was detected by real-time fluorescence quantitative PCR. The changes of NF-κBp65 and caspase-1p20 activities in Hela cells were detected by Western Blot. The levels of interleukin-1β IL-1β) content. Results In Hela cells, the expression of NALP3mRNA, NF-κB and caspase-1p20 activity and IL-1βin cell culture supernatant were significantly increased under the stimulation of Neisseria gonorrhoeae, there were statistically significant differences between Ng group and blank control group (P <0.01). After the intervention of progesterone, the content of NALP3mRNA, the activity of NF-κB and caspase-1p20, the content of IL-1β in cell culture supernatant decreased obviously with the increase of progesterone concentration, Compared with Ng group, the difference was statistically significant (P all <0.01). Conclusion Progesterone can inhibit Neisseria gonorrhoeae-induced NALP3 expression and caspase-1 activation and IL-1β secretion, and this inhibition may be through inhibition of NF-κB activity.