孕激素对小鼠脾细胞淋球菌感染模型NLRP3炎性体表达及Th17/Treg分化的影响  

Effects of progesterone on the expression of NLRP3 and Th17/Treg differentiation in murine splenocyte gonococcal infection model

作　　者：许莉[1] 陈娜[1] XU Li;CHEN Na(Department of Dermatology,Wuhan No.1 Hospital,Wuhan 430022,China)

机构地区：[1]武汉市第一医院皮肤科,湖北武汉430022

出　　处：《南京医科大学学报（自然科学版）》2021年第11期1574-1578,1591,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：武汉市科技局应用基础前沿项目(2020020601012312);武汉市卫生健康委科研计划资助项目(WX19D28,WX15B13)。

摘　　要：目的:研究孕激素对小鼠脾细胞淋球菌(Neisseria gonorrhoeae,Ng)感染模型中NLRP3炎性体表达及Th17/Treg分化的影响。方法:体外分离、培养小鼠脾细胞后,分为如下5组:空白对照组、Ng组、Ng+1×10^(-9) mol/L孕酮组、Ng+1×10^(-8) mol/L孕酮组、Ng+1×10^(-7) mol/L孕酮组。培养3 d后收集细胞,采用流式细胞术检测Th17/Treg细胞比例,利用实时荧光定量PCR检测各组细胞NLRP3、Th17/Treg特异性转录因子RORγt、Foxp3 mRNA表达水平,Western blot检测各组细胞NLRP3、Caspase-1p20、RORγt、Foxp3蛋白表达。ELISA法检测脾细胞培养上清中白细胞介素(interleukin,IL)-1β及Th17/Treg相关细胞因子IL-17、转化生长因子(transforming growth facfor,TGF)-β、IL-10水平。结果:脾细胞在淋球菌的刺激下,Th17细胞和Treg细胞比例增多,细胞内NLRP3、Th17/Treg特异性转录因子RORγt、Foxp3 mRNA和蛋白表达水平升高,Caspase-1p20蛋白表达水平升高,细胞培养上清中IL-1β、IL-17、TGF-β、IL-10含量增高(P<0.01);加入孕酮预处理后,随着孕酮作用浓度的增高,Th17细胞比例、细胞内NLRP3和RORγt的mRNA和蛋白表达水平、Caspase-1p20蛋白表达水平、细胞培养上清中IL-1β和IL-17含量均明显降低,Treg细胞比例、细胞内Foxp3 mRNA和蛋白表达、细胞培养上清中TGF-β和IL-10含量均明显增高,与Ng组比较差异有统计学意义(P<0.01)。结论:孕激素可抑制Ng感染诱导的小鼠脾细胞NLRP3表达及Th17细胞分化,促进Ng感染诱导的Treg细胞分化。Objective:This study aims to investigate the effects of progesterone on the expression of NLRP3 and the differentiation of Th17/Treg in murine splenocytes infected by Neisseria gonorrhoeae(Ng).Methods:Murine splenocytes were isolated in vitro,and randomly divided into five groups:control group,Ng group,Ng+1×10^(-9)mol/L progesterone group,Ng+1×10^(-8)mol/L progesterone group,Ng+1×10^(-7)mol/L progesterone group.The splenocytes were collected after 3 days of culture,the proportions of Th17/Treg cells were detected by intracellular staining using flow cytometry.The mRNA expressions of NLRP3,Th17/Treg-specific transcription factors RORγt and Foxp3 in murine splenocytes were detected by real-time PCR.The protein expression levels of NLRP3,Caspase-1 p20,RORγt and Foxp3 in murine splenocytes were determined by Western blot.The concentrations of interleukin(IL)-1β and Th17/Treg related cytokines IL-17,transforming growth facfor(TGF)-β,IL-10 in the supernatant were detected by ELISA.Results:Under the stimulation of Ng,the proportions of Th17 cells and Treg cells,the mRNA and protein levels of NLRP3,Th17/Treg-specific transcription factors RORγt and Foxp3 in the splenocytes,the protein levels of Caspase-1 p20 and the concentrations of IL-1β,IL-17,TGF-β and IL-10 in the supernatant increased than those in the controls(P<0.01).After pretreated with progesterone,the proportion of Th17 cells,the m RNA and protein levels of NLRP3 and RORγt in the splenocytes,the protein levels of Caspase-1 p20 and the concentrations of IL-1β and IL-17 in the supernatant were significantly lower than those in Ng group.The proportion of Treg cells,the mRNA and protein levels of Foxp3 in the splenocytes and the concentrations of TGF-β and IL-10 in the supernatant were significantly higher than those in Ng group(P<0.01).Conclusion:Progesterone can inhibit the expression of NLRP3 and Th17 differentiation,and promote the differentiation of Treg cells in murine splenocytes induced by Ng infection.

关 键 词：淋球菌 孕激素 NLRP3 TH17/TREG 

分 类 号：R392.12[医药卫生—免疫学]