NF-κB p65/TNF-α对PPROM孕妇胎膜细胞凋亡的影响  

Effect of NF-κB p65/TNF-α on the apoptosis of fetal membrane cell in pregnantwomen with PPROM

作　　者：唐石欢 肖玲 李细平 曾莉 周仁慧 李恒 阳孝明 张旋 胡琼 张红梅 张先平 Tang Shihuan;Xiao Ling;Li Xiping;Zeng Li;Zhou Renhui;Li Heng;Yang Xiaoming;Zhang Xuan;Hu Qiong;Zhang Hongmei;Zhang Xianping(Department of Obstetrics and Gynecology,the Affiliated Loudi Hospital,Hengyang Medical School,University of South China,Loudi Hunan 417000,P.R.China)

机构地区：[1]南华大学附属娄底医院(南华大学衡阳医学院)妇产科,湖南娄底417000

出　　处：《中国计划生育和妇产科》2023年第12期92-96,101,I0004,共7页Chinese Journal of Family Planning & Gynecotokology

基　　金：娄底市科技局重大专项项目(项目编号:娄财教指2022[2]号)。

摘　　要：目的探讨核转录因子kappa B p65(nuclear factor-kappa B p65,NF-κB p65)/肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)对未足月胎膜早破(preterm premature rupture of membranes,PPROM)孕妇胎膜细胞凋亡的影响。方法选取2021年6月至2022年6月于南华大学附属娄底医院分娩的57例孕妇为研究对象,PPROM孕妇27例为试验组,正常足月分娩孕妇30例为对照组,采集分娩时胎膜组织。利用HE染色法观察胎膜组织结构;免疫组织化学法检测胎膜NF-κB p65和磷酸化NF-κB p65(p-NF-κB p65)的表达;TUNEL检测胎膜细胞凋亡;Western Blotting检测胎膜NF-κB p65、p-NF-κB p65、TNF-α和半胱氨酸天冬氨酸蛋白水解酶-3(cysteinyl aspartate specific proteinase-3,Caspase-3)蛋白表达水平;qRT-PCR检测胎膜组织NF-κB p65、TNF-α和Caspase-3 mRNA表达;分析PPROM孕妇胎膜p-NF-κB p65/NF-κB p65比值、TNF-α蛋白表达水平与胎膜细胞凋亡指数的相关性。结果HE染色结果显示对照组胎膜组织结构较完整,试验组胎膜组织结构排列紊乱、胶原纤维层变薄、细胞胞质呈现大量空泡化;免疫组化结果显示NF-κB p65主要表达于胎膜细胞的细胞质,p-NF-κB p65表达于胎膜细胞的细胞核;试验组胎膜NF-κB p65、TNF-α、Caspase-3蛋白和mRNA表达以及细胞凋亡指数均高于对照组,差异有统计学意义(P<0.05);试验组p-NF-κB p65/NF-κB p65蛋白比值高于对照组(P<0.01);试验组p-NF-κB p65/NF-κB p65蛋白比值与细胞凋亡指数呈正相关(相关系数r_1=0.897,P<0.01);试验组p-NF-κB p65/NF-κB p65与TNF-α蛋白表达水平呈正相关(r_2=0.835,P<0.01);试验组TNF-α蛋白表达水平与胎膜细胞凋亡指数呈正相关(r_3=0.934,P<0.01)。结论NF-κB p65可通过促进下游基因TNF-α表达,进而上调Caspase-3,引起胎膜细胞凋亡,参与PPROM的发生。Objective To investigate the effect of nuclear factor-kappa B p65(NF-κB p65)/tumor necrosis factor-α(TNF-α)on the apoptosis of fetal membrane cells in pregnant women with preterm premature rupture of membranes(PPROM).Methods 57 pregnant women who delivered in the Affiliated Loudi Hospital,University of South China from June 2021 to June 2022 were selected as the research objects.27 PPROM pregnant women were included in the experimental group,and 30 pregnant women with normal term delivery were included in the control group,and fetal membrane tissue was collected at delivery.The tissue structure of the fetal membrane was observed by HE staining.Immunohistochemical detection of fetal membrane NF-κB p65 and phosphorylated NF-κB p65(p-NF-κB p65).TUNEL was applied to detect cell apoptosis in fetal membrane tissue.The expression of NF-κB p65,p-NF-κB p65,TNF-α and cysteinyl aspartate specific proteinase-3(Caspase-3)protein expression levels were detected by Western Blotting.The mRNA expression of NF-κB p65,TNF-α and Caspase-3 in fetal membrane tissue was detected by qRT-PCR.The correlation between the expression level of p-NF-κB p65/NF-κB p65,TNF-α protein and the fetal cell apoptosis index in PPROM was analyzed.Results HE staining showed that the structure of fetal membrane tissue in control group was complete,while the structure of fetal membrane tissue was disordered,the collagen fiber layer was thin,and the cytoplasm showed largely vacuolation in the experimental group.Immunohistochemical staining showed that the NF-κB p65 was mainly expressed in the cytoplasm of fetal membrane cells,and p-NF-κB p65 was mainly expressed in the nucleus of fetal membrane cells.The expression level of NF-κB p65,TNF-α,Caspase-3 proteins and mRNA,and the cell apoptosis index in the experimental group were higher than those in the control group(P<0.05).The p-NF-κB p65/NF-κB p65 protein ratio in the experimental group were significantly higher than that in the control group(P<0.01).In experimental group,the p-NF-κB p65

关 键 词：未足月胎膜早破 NF-κB p65 TNF-Α CASPASE-3 凋亡 

分 类 号：R714.433[医药卫生—妇产科学]