缺氧复氧诱导自噬基因表达对滋养细胞生长的影响与机制

发布时间：2018-07-14 15:34

【摘要】：为探索子痫前期孕妇胎盘病变的发病机制,我们模拟体内缺血再灌注微环境,在体外建立胎盘滋养细胞HTR8/SVneo缺氧复氧模型,以探究缺氧复氧对细胞自噬的诱导作用及对细胞生长的影响。将实验分为对照组、缺氧复氧组及自噬抑制剂3-MA+缺氧复氧组,应用吖啶橙染色及LC3-Ⅱ免疫荧光染色检测经处理24 h后细胞自噬水平,MTT法检测细胞增殖能力,Real time PCR检测自噬基因Beclin-1、LC3-Ⅱ的表达,Western blot分析相应自噬蛋白的表达。结果显示缺氧复氧组HTR8/SVneo细胞自噬水平明显升高(p0.01),伴随Beclin-1、LC3-Ⅱ基因及蛋白表达显著增高(p0.01),细胞增殖同时显著受抑(p0.01),而加入3-MA后缺氧复氧组细胞自噬水平明显受抑(p0.01),Beclin-1、LC3-Ⅱ基因及蛋白表达显著下降(p0.01),细胞增殖能力显著提高(p0.01)。这表明滋养细胞HTR8/SVneo在缺氧复氧环境中启动细胞自噬,过度自噬时可能通过诱导Ⅱ型程序性死亡影响细胞增殖,当自噬被抑制后,细胞增殖能力明显恢复。
[Abstract]:In order to explore the pathogenesis of placental lesions in preeclampsia, we simulated the microenvironment of ischemia reperfusion in the body and established the HTR8/SVneo hypoxia reoxygenation model in vitro to explore the effect of hypoxia reoxygenation on the induction of autophagy and the effect on cell growth. The experiment was divided into the control group, the hypoxia reoxygenation group and the autophagy inhibitor 3-. MA+ hypoxia reoxygenation group, using acridine orange staining and LC3- II immunofluorescence staining to detect the autophagy level of cells after treatment 24 h, MTT method to detect cell proliferation, Real time PCR detection of autophagic gene Beclin-1, LC3- II expression, Western blot analysis of the corresponding autophagic protein expression. Results showed the level of autophagy in the hypoxia reoxygenation group. Significantly increased (P0.01), with Beclin-1, LC3- II gene and protein expression increased significantly (P0.01), cell proliferation was significantly inhibited (P0.01), but after the addition of 3-MA, the level of autophagy in the anoxic reoxygenation group was significantly inhibited (P0.01), Beclin-1, LC3- II gene and protein expression significantly decreased (P0.01), cell proliferation ability significantly increased (P0.01). This indicates that the proliferation of cells (P0.01). The cultured cell HTR8/SVneo initiating autophagy in the anoxic reoxygenation environment may affect cell proliferation by inducing type II programmed death. When autophagy is suppressed, the cell proliferation ability is obviously restored.
【作者单位】：深圳市妇幼保健院产科;
【基金】：广东省医学科研基金项目(B2013354)资助
【分类号】：R714.244

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