Page 12 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第41卷第11期
·1574 · Journal of Nanjing Medical University（Natural Sciences） 2021年11月

·基础研究·

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

许 莉 ，陈 娜
*
武汉市第一医院皮肤科，湖北 武汉 430022

［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 mol/L孕酮组、Ng+1×10 mol/L孕酮
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组、Ng+1×10 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
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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。
［关键词］ 淋球菌；孕激素；NLRP3；Th17/Treg
［中图分类号］ R392.12 ［文献标志码］ A ［文章编号］ 1007⁃4368（2021）11⁃1574⁃06
doi：10.7655/NYDXBNS20211102

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

*
XU Li ，CHEN Na
Department of Dermatology，Wuhan No.1 Hospital，Wuhan 430022，China

［Abstract］ 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 mol/L progesterone group，Ng+1×10 mol/L progesterone group，
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Ng+1×10 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⁃1p20，
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⁃1p20 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

［基金项目］ 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）
∗
通信作者（Corresponding author），E⁃mail：rita_76@163.com