高糖诱导Txnip及氧化应激反应对妊娠期糖尿病胎盘浸润力的作用及机制探讨

Gestational diabetes mellitus (GDM) seriously endangers the health of mother and child. With the development of society, the incidence of GDM is increasing year by year, but its pathogenesis of GDM is not clear. In clinic, it only can control glocemic index, still unable to prevent and treat GDM actively. Previous studies found that hyperglycemia stimulates placental trophoblast cells and can induce increased expression of mRNA and protein in Txnip, which aggravates oxidative stress reaction and increases apoptosis. In clinical sample, the expression of Txnip in the placenta of GDM patients is significantly higher than that of normal placenta. This project takes human placental trophoblast cells as a research object to establish a GDM placental cell model and studies the effect of Txnip genes on oxidative stress reaction, apoptosis and placental trophoblast cell characteristics (such as invasiveness and transference) through CO-IP, FACS and other techniques. It is to reveal the relationship between the hyperglycemia environment and placental trophoblast invasiveness and its damage, and explore the molecular mechanism of Txnip-Ros signal pathway in placental trophoblast and placenta formation and development in GDM. The project is expected to find the molecular mechanism of Txnip-Ros signal pathway in hyperglycemia induced damage of placentaltrophoblast cells, clarify the effect of Txnip onthe damage of GDMinplacentadevelopemnt, and provide experimental and theoretical basis for prevention and targeted treatment of GDM.

妊娠期糖尿病（GDM）严重危害母婴健康。随着社会发展，GDM的发病率逐年上升，但其发病机制尚不明确，临床上只能控制血糖，还无法进行主动预防和治疗。前期研究发现高糖刺激胎盘细胞，可诱导Txnip的mRNA和蛋白表达升高，加剧氧化应激反应及增加细胞凋亡，临床标本中发现，GDM患者胎盘中Txnip的表达高于正常胎盘。鉴于此，本项目以人胎盘滋养层细胞为研究对象建立GDM胎盘细胞模型，通过CO-IP、FACS等技术，研究Txnip基因对细胞氧化应激、细胞凋亡及滋养层细胞特性（如浸润、迁移等）的影响，揭示高糖环境与胎盘滋养层细胞浸润力变化及其损伤的关系，探索GDM中Txnip-Ros信号通路在胎盘滋养层以及胎盘的形成、发育过程中的分子机制。本项目预期发现Txnip-Ros信号通路在高糖诱导胎盘滋养层细胞损伤的分子机制，阐明Txnip在GDM胎盘发育损伤的影响，为GDM的预防和靶向治疗提供实验理论依据。

前言：我们前期研究已证实妊娠期糖尿病(GDM)患者胎盘中葡萄糖敏感基因-硫氧还蛋白互作用蛋白(TXNIP)表达升高，但TXNIP表达对妊娠期糖尿病(GDM)胎盘的影响及机制尚不清楚。在本研究中，我们通过在体外引入Tet-on“开关”模拟TXNIP的高表达，接近其在现实世界中的表达水平，探讨TXNIP异常的后续后果。方法:采用免疫荧光、Western blot和RT-QPCR方法研究TXNIP在GDM患者和健康对照组胎盘中的表达和定位。将Tet-on系统转染人胎盘滋养层细胞HTR -8/Svneo细胞，实现TXNIP的过表达。通过CRISPR/Cas9方法获得TXNIP基因敲除细胞系，通过IncuCyte成像系统和流式细胞仪观察细胞表型。结果:TXNIP在GDM胎盘组织中的表达比对照组高近2倍，TXNIP的位置在胎盘滋养层细胞。TXNIP表达升高时，诱导细胞迁移和侵袭，但对凋亡和增殖无明显改变。此外，Tet-on TXNIP细胞体积变大，Vimentin和E-cadherin表达升高，N-cadherin表达降低。STAT3在Tet-on TXNIP细胞中的表达及磷酸化水平较高。在KO-TXNIP细胞中观察到相反的上述变化。.结论:TXNIP异常表达可能与GDM胎盘功能受损有关，并通过STAT3和EMT相关通路影响胎盘滋养细胞的迁移和侵袭，因此，TXNIP可能是修复GDM患者胎盘功能缺陷的潜在治疗靶点。