TFDP2在子痫前期胎盘中的表达及其对滋养细胞凋亡的影响  

Expression of TFDP2 in placentas of women with preeclampsia and its effect on apoptosis of trophoblast cells

作　　者：曹晨睿 刘丹 王志尹 赵光锋[1] 裴中锐 胡娅莉[1] Cao Chenrui;Liu Dan;Wang Zhiyin;Zhao Guangfeng;Pei Zhongrui;Hu Yali(Department of Obstetrics and Gynecology,Nanjing Drum Tower Hospital,the Affiliated Hospital of Nanjing University Medical School,Nanjing 210009,China)

机构地区：[1]南京大学医学院附属鼓楼医院妇产科,南京210009

出　　处：《中华围产医学杂志》2024年第2期133-142,共10页Chinese Journal of Perinatal Medicine

基　　金：国家重点研发计划(2021YFC2701603);国家自然科学基金(82271716);南京市卫生科技发展专项资金杰出青年基金(JQX21004)。

摘　　要：目的探讨子痫前期患者胎盘组织中转录因子二聚化配体2(transcription factor dimerization partner 2,TFDP2)的表达及其对滋养细胞凋亡的影响。方法回顾性选取2018年1月至2022年12月期间在南京大学医学院附属鼓楼医院剖宫产分娩的子痫前期产妇30例(子痫前期组),及同期在本院剖宫产分娩的健康正常产妇30例(对照组)的胎盘组织。利用免疫组织化学染色检测TFDP2在胎盘组织中的定位,实时荧光定量-聚合酶链反应和蛋白质印迹技术检测2组胎盘组织中TFDP2 mRNA和蛋白水平的差异。取合体滋养细胞BeWo细胞,转染小干扰RNA敲降TFDP2,利用蛋白质印迹和实时荧光定量-聚合酶链反应技术检测凋亡相关指标——B细胞淋巴瘤2蛋白(B cell lymphoma 2,Bcl2)和Bcl2相关X蛋白(Bcl2 associated X,Bax)及其mRNA的改变,利用末端脱氧核苷酸转移酶介导的dUTP原位切口末端标记染色和流式细胞技术观察凋亡细胞数量的改变,并探索相关下游信号通路的变化。采用两独立样本t检验、秩和检验和χ^(2)检验进行统计学分析。结果TFDP2主要表达于对照组胎盘合体滋养细胞和绒毛外滋养细胞中。子痫前期胎盘组织合体滋养细胞中TFDP2的表达量在mRNA水平(0.722±0.239与1.000±0.348,t=3.61,P=0.001)和蛋白水平(0.728±0.185与1.000±0.206,t=2.41,P=0.037)均低于对照组。与未敲降TFDP2比较,在BeWo细胞中敲降TFDP2,凋亡指标Bax/Bcl2比值升高(mRNA:1.755±0.452与1.000±0.279,t=3.48,P=0.006;蛋白:3.206±0.922与1.000±0.290,t=3.95,P=0.017),每个视野中凋亡细胞数量升高[(4.556±1.740)与(2.444±1.130)个,t=3.05,P=0.008],总凋亡细胞比例升高[(21.37±1.66)%与(12.61±0.38)%,t=8.92,P=0.001],BeWo细胞质中蛋白激酶A催化亚基的Thr197位点磷酸化活性降低(0.466±0.035与1.000±0.075,t=11.19,P<0.001),细胞核中β-连环蛋白的表达降低(0.250±0.093与1.000±0.269,t=4.57,P=0.010)。结论子痫前期患者胎盘组织合体滋养细胞中TFDPObjectiveTo investigate the expression level of transcription factor dimerization partner 2(TFDP2)in the placentas of women with preeclampsia,and analyze its effect on the apoptosis of trophoblast cells.MethodsPlacental tissues from thirty puerperae with preeclampsia who gave birth by cesarean section in Nanjing Drum Tower Hospital,the Affiliated Hospital of Nanjing University Medical School between January 2018 and December 2022(preeclampsia group)and 30 healthy puerperae undergoing cesarean section during the same period(control group)were retrospectively selected.Immunohistochemistry was used to localize TFDP2 in the placental tissues.Real-time quantitative-polymerase chain reaction(qRT-PCR)and Western blot were used to detect the differences in expression of TFDP2 at mRNA and protein levels in placental tissues between the two groups.Forskolin-exposed BeWo cells were transfected with small interfering RNA(siRNA)to knockdown TFDP2 and the changes in the expression of apoptosis-related indicators,B cell lymphoma 2(Bcl2)and Bcl2 associated X(Bax),at protein and mRNA levels were analyzed by Western blot and qRT-PCR,respectively.Besides,the change in the apoptosis level of BeWo cells was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining and flow cytometry.Downstream signaling pathways were analyzed to understand the involved molecular mechanisms.Two independent samples t-test,Wilcoxon rank-sum test,and Chi-square test were used for statistical analysis.ResultsTFDP2 was mostly localized in the syncytiotrophoblasts and the extravillous trophoblasts in the normal placentas.TFDP2 expression in the syncytiotrophoblasts was lower in the preeclampsia group than in the control group at both mRNA(0.722±0.239 vs.1.000±0.348,t=3.61,P=0.001)and protein(0.728±0.185 vs.1.000±0.206,t=2.41,P=0.037)levels.Comparing the group without knockdown of TFDP2,the knockdown of TFDP2 in BeWo cells elevated the Bax/Bcl2 ratio(mRNA:1.755±0.452 vs.1.000±0.279,t=3.48,P=0.006;protein:3.206�

关 键 词：先兆子痫 DNA结合蛋白质类 转录因子 滋养层 细胞凋亡 

分 类 号：R714.244[医药卫生—妇产科学]