摘要
目的探讨早发型子痫前期(early-onset preeclampsia, EOPE)和晚发型子痫前期(late-onset preeclampsia, LOPE)胎盘组织中自噬和凋亡水平的差异,利用人绒毛膜滋养层细胞株HTR 8/SVneo缺氧模型探讨自噬与凋亡的关系,为解释子痫前期(preeclampsia, PE)的发病机制提供新的思路。方法选取2018年1~6月上海交通大学医学院附属国际和平妇幼保健院产科剖宫产分娩孕妇26例的临床资料。按照PE发生程度分为EOPE组10例,LOPE组6例,正常孕妇组10例。通过透射电镜观察EOPE组、LOPE组和正常孕妇组胎盘组织滋养层细胞中的自噬溶酶体;采用实时荧光定量多聚核苷酸链式反应和免疫印迹方法检测EOPE组、LOPE组和正常孕妇组胎盘组织中自噬相关基因(LC 3和Beclin-1)、抗凋亡基因Bcl-2和促凋亡基因Bax mRNA和蛋白水平的表达;利用HTR 8/SVneo缺氧模型,检测LC 3、Beclin-1、Bcl-2和Bax mRNA水平的表达。结果与正常孕妇组胎盘组织相比,EOPE组自噬溶酶体含量降低,LC 3、Beclin-1和Bcl-2 mRNA和蛋白表达水平降低,Bax表达升高,差异均有统计学意义(P<0.05)。LOPE组自噬溶酶体含量增加并且出现凋亡细胞,LC 3、Beclin-1和Bax表达均升高,Bcl-2表达降低,差异均有统计学意义(P<0.05)。缺氧模型中,与正常HTR 8/SVneo细胞相比,缺氧12 h组LC 3和Beclin-1 mRNA表达水平均升高,Bcl-2和Bax mRNA表达水平均降低,差异有统计学意义(P<0.05),抑制自噬活性后,Bax mRNA表达水平显著升高,差异有统计学意义(P<0.05);24 h和48 h组LC 3、Beclin-1、Bcl-2和Bax的mRNA表达均降低,差异有统计学意义(P<0.05);72 h组LC 3、Beclin-1和Bax表达均升高,Bcl-2表达降低,差异有统计学意义(P<0.05)。结论 EOPE组胎盘滋养层细胞中自噬活性受损,凋亡水平增加;而LOPE组中自噬和凋亡水平均增加;短期缺氧激活滋养层细胞自噬,凋亡降低,抑制自噬凋亡增加;长期缺氧引发过度自噬和凋亡。
Objective To investigate the differences of trophoblastic autophagy and apoptosis between early-onset preeclampsia(EOPE)and late-onset preeclampsia(EOPE)and to explore the relationship between autophagy and apoptosis in the human trophoblast HTR 8/SVneo cell line under hypoxia condition, so as to provide new ideas for explaining the pathophysiology mechanism of the development of preeclampsia. Methods From Jannary 2018 to June 2018,26 pregnoncy nomen with cesarean section were selected from the International Peace Maternity and Child Health Hospital Affiliated to the School of Medicine of Shanghai Jiaotong University. According to the occurrence degree of PE,10 cases were devided into EOPE group,6 cases in LOPE group and 10 cases in normal pregnancy women group.The autolysosomes and apoptosis bodies in placenta tissues of EOPE and LOPE were elucidated by transmission electron microscopy;qRT-PCR and Western blot were used to detect the mRNA and protein expression of LC 3, Beclin-1, Bcl-2 and Bax in placenta tissues. Results The results showed that compared with normal placenta, the number of autolysosomes in EOPE group was reduced, the expressions of LC 3, Beclin-1 and Bcl-2 were decreased at the level of mRNA and protein, and the expression of Bax was increased(P<0.05);The number of autolysosomes and the expression of LC 3, Beclin-1 and Bax were increased in LOPE group, the expression of Bcl-2 was decreased(P<0.05). In cells model, the mRNA expression of LC 3 and Beclin-1 was increased compared with normal HTR 8/SVneo cells in the hypoxia group at 12 h, while the mRNA expression of Bcl-2 and Bax was deceased(P<0.05);The mRNA expression of Bax was significantly increased when autophagy inhibiting(P<0.05);In the hypoxia groups at 24 h and 48 h, both of the mRNA expression of LC 3, Beclin-1, Bcl-2 and Bax were decreased compared with control group(P<0.05). The mRNA expression of LC 3, Beclin-1 and Bax was increased in hypoxia group at 72 h while the expression of Bcl-2 was decreased(P<0.001). Conclusion There are im
作者
李亚梅
杨星宇
赵雪娅
程蔚蔚
LI Ya-mei;YANG Xing-yu;ZHAO Xue-ya;CHENG Wei-wei(Department of Obstetrics and Gynecology,The International Peace Maternity and Child Health Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai Key Laboratory of Embryo Original Diseases,Shanghai Municipal Key Clinical Specialty,Shanghai 200030,P.R.China)
出处
《中国计划生育和妇产科》
2019年第11期38-43,I0001,共7页
Chinese Journal of Family Planning & Gynecotokology
基金
国家自然科学基金资助项目(项目编号:81501256)
上海市卫生和计划生育委员会资助项目(项目编号:201640361)
徐汇区市卫生规划委员会重大疾病项目(项目编号:XHLHGG201805)
