高糖环境下beclin2介导的自噬增强绒毛膜滋养层细胞凋亡  

High glucose induces apoptosis of chorionic trophoblast cells via beclin2-mediated autophagy

作　　者：纪璐璐[1] 熊国平[2] 汪琳[1] Ji Lulu;Xiong Guoping;Wang Lin(Department of Histology and Embryology, School of Basic Medical Sciences,Wuhan University, Wuhan 430071, China;Department of Gynaecology and Obstetrics, The Central Hospital of Wuhan, Wuhan 430014, China)

机构地区：[1]武汉大学基础医学院组织学与胚胎学教研室,武汉430071 [2]武汉市中心医院妇产科,武汉430014

出　　处：《中国组织化学与细胞化学杂志》2017年第2期109-114,共6页Chinese Journal of Histochemistry and Cytochemistry

摘　　要：目的探讨高糖对人绒毛膜滋养层细胞早期凋亡及自噬水平的影响。方法取足月妊娠的正常及妊娠期糖尿病(gestational diabetes mellitus,GDM)患者的胎盘组织,透射电镜观察滋养层细胞超微结构的改变。体外用不同浓度的含糖培养基培养滋养层细胞系HTR8/Svneo细胞24h,分为低糖组(1.57mmol/L或LG2 2.25mmol/L),正常对照组(5.57mmol/L)和高糖组(10.57mmol/L、15.57mmol/L或40.57mmol/L);流式细胞术检测细胞早期凋亡率;q-PCR检测细胞内beclin1、beclin2和ATG7等自噬相关基因的m RNA表达水平;Western blot检测Ⅱ型自噬标志蛋白微管相关蛋白1轻链3(LC3-II)和p62蛋白表达水平。结果透射电镜下可见GDM组滋养层细胞中存在自噬小体且空泡数目明显多于正常足月妊娠组,其游离面微绒毛出现明显倒覆及坍塌现象;在体外培养的滋养层细胞中,流式细胞术检测显示1.57mmol/L低糖组及40.57mmol/L高糖组较正常对照组HTR8/SVneo细胞的早期凋亡率均明显增加;q-PCR分析发现40.57mmol/L高糖组beclin2及ATG7 m RNA表达水平较正常对照组升高,beclin1 m RNA表达无差异;Western blot检测表明,与正常对照组比较,40.57mmol/L高糖组LC3-II蛋白表达水平升高,p62蛋白表达水平降低。结论高糖可通过beclin2介导的自噬信号途径增强滋养层细胞自噬水平进而导致细胞凋亡,并可能与不良的妊娠结局相关。Objective To investigate the effect of high glucose on the early apoptosis and autophagy of human chorionic trophoblast cells.Methods Full-term placenta tissue was sampled from normal cases and gestational diabetes mellitus(GDM)cases,and the changes in the ultrastructure of trophoblast cells were observed by transmission electron microscopy.HTR8/SVneo trophoblast cells were cultured in vitro in low,normal and high glucose conditions(LG1:1.57mmol/L,LG2:2.25mmol/L,NC:5.57mmol/L,HG1:10.57mmol/L,HG2:15.57mmol/L,HG3:40.57mmol/L)for24hours.The early apoptosis rate was measured by flow cytometry.The mRNA levels of autophagy-related genes Beclin2,ATG7and Beclin1were analyzed by qPCR.The levels of LC3-II and p62protein were detected by western blot.Results Autophagosomes were observed by transmission electron microscopy in trophoblast cells in the GDM group,as well as an increased number of vacuoles compared with that of the normal group.Remarkable collapse of microvilli on the free surface was also observed in the GDM group.The flow cytometry results showed that the early apoptosis rate was significantly higher in groups LG1and HG3than in group NC.The qPCR test displayed that the levels of beclin2and ATG7mRNA increased obviously in group HG3,while that of beclin1mRNA showed no significant difference from group NC.The western blot results showed that the level of LC3-II protein was higher and that of p62lower in group HG3than in group NC.Conclusion High glucose promotes the autophagy of trophoblast cells via Beclin2-mediated signaling,which results in cell apoptosis and may lead to adverse fetal outcomes.

关 键 词：妊娠期糖尿病 HTR8/SVneo 自噬 凋亡 beclin2 

分 类 号：R329.1[医药卫生—人体解剖和组织胚胎学]