脂联素通过AdipoR2-AMPK-内质网应激-凋亡途径对高糖环境下滋养细胞损伤的保护作用及临床意义

项目名称： 脂联素通过AdipoR2-AMPK-内质网应激-凋亡途径对高糖环境下滋养细胞损伤的保护作用及临床意义

项目编号： No.81501258

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 韩云

作者单位： 南通大学

项目金额： 17万元

中文摘要： 本研究以妊娠期糖尿病胎盘滋养细胞存在过度凋亡和脂联素的抗凋亡作用为切入点，通过研究脂联素对妊娠期糖尿病孕妇胎盘滋养细胞及体外滋养细胞系Bewo细胞凋亡的影响，探讨脂联素通过AdipoR2-AMPK-内质网应激-凋亡途径对高糖环境下滋养细胞损伤的保护作用。（1）建立滋养细胞系Bewo高糖细胞模型，采用透射电镜、流式细胞仪、RT-PCR和Western blot技术检测孕妇及体外滋养细胞的内质网应激及凋亡情况；（2）观察分析妊娠期糖尿病胎盘滋养细胞内质网应激、凋亡与妊娠结局的关系；（3）探讨脂联素通过减轻内质网应激减轻滋养细胞凋亡，对滋养细胞起保护作用；（4）给予AMPK抑制剂阿糖腺苷(AraA）和小的干扰RNA抑制AdipoR2，探讨脂联素通过脂联素受体R2激活细胞内第二信使AMPK，让它参与内质网应激调控滋养细胞凋亡；最终为监测和改善妊娠期糖尿病妊娠结局提供有意义的实验依据和治疗的新靶点。

中文关键词： 妊娠期糖尿病；胎盘；脂联素；内质网应激；细胞凋亡

英文摘要： This study is based on the theory of the excessive apoptosis in placental cytotrophoblasts of gestational diabetes mellitus and antiapoptosis property of adiponectin. The experiments focus on identifying the mechanism how adiponectin protects injured cytotrophoblasts under high glucose environment through AdipoR2 - AMPK - endoplasmic reticulum stress response - apoptosis pathway by researching on the effect of adiponectin on the apoptosis of cytotrophoblasts from pregnant women with gestational diabetes mellitus and cell line Bewo cultured in vitro. The steps of this study are designed as below: (1) Culture the cytotrophoblast cell line Bewo under high glucose environment to build up the cell model. Observe the changes of endoplasmic reticulum stress and apoptosis of cytotrophoblast in placenta with gestational diabetes mellitus and under high glucose environment in vitro by transmission electron microscopy, flow cytometry instrument, RT-PCR and Western blot technique; (2) Clarify the association of the different endoplasmic reticulum stress and apoptosis with pregnancy outcomes in gestational diabetes mellitus; Clarify how adiponectin protects the cytotrophoblast by reducing the endoplasmic reticulum stress response and slowing down the apoptosis process; (4) AMPK inhibitor AraA and siRNA to suppress AdipoR2 are utilized, to identify if adiponectin activates the intracellular second messenger AMPK through AdipoR2 to participate in endoplasmic reticulum stress to regulate the apoptosis of cytotrophoblast. Thus, the results will be used to monitor and improve pregnancy outcomes of gestational diabetes mellitus, and lay a foundation of eugenic and superior nurture in our country.

英文关键词： Gestational diabetes mellitus ;placenta ;adiponectin;endoplasmic reticulum stress;apoptosis