内质网应激通路参与不同缺氧条件下主动脉内皮细胞凋亡的研究

发布时间：2018-05-26 19:46

本文选题：缺氧 + 内质网应激　；参考：《华中科技大学》2013年硕士论文

【摘要】：目的：观察不同缺氧条件下内皮细胞凋亡情况及内质网应激因子表达变化情况，探讨内质网应激通路参与不同缺氧条件下主动脉内皮细胞凋亡的相关机制。方法： 24只SPF级wistar大鼠随机分成3组，每组8只（n=8）：常氧对照组（NC组）、间歇缺氧组（IH组）、持续缺氧组（PH组）。通过免疫组织化学方法检测主动脉弓处内皮细胞中内质网应激信号分子BIP、CHOP、caspase12的蛋白表达，TUNEL法检测主动脉弓处内皮细胞凋亡情况；体外培养人脐静脉内皮细胞，随机分组：常氧对照组（NC1组）、常氧对照+抑制剂组（NC1+TUDCA组）、间歇缺氧组（IH1组）、间歇缺氧+抑制剂组（IH1+TUDCA组）、持续缺氧4H组（PH1组）、持续缺氧4H+抑制剂组（PH1+TUDCA组）、持续缺氧12H组（PH2组）、持续缺氧12H+抑制剂组（PH2+TUDCA组）。通过流式细胞术检测各组细胞凋亡情况，real-time PCR检测各组细胞内质网应激因子BIP、CHOP、caspase12的mRNA表达水平变化。结果：体内、外研究结果均显示，与正常对照组相比，间歇缺氧组及持续缺氧组内皮细胞凋亡率增加（P0.05），与此同时，内质网应激因子BIP、CHOP、caspase12表达明显增高（P0.05），其中，同等时长下，，间歇缺氧组内皮细胞凋亡率及内质网应激因子表达增高较持续缺氧组明显（P0.05）。加入内质网应激抑制剂TUDCA后，内皮细胞凋亡率和内质网应激因子BIP、CHOP、caspase12表达明显下降（P0.05）。结论：缺氧（尤其是间歇缺氧）能导致主动脉内皮细胞凋亡，内质网应激通路可能是介导缺氧诱导主动脉细胞凋亡过程中一条很重要的途径。
[Abstract]:Objective: To observe the changes of endothelial cell apoptosis and endoplasmic reticulum stress factor expression under different hypoxia conditions, and to explore the mechanism of endoplasmic reticulum stress pathway involved in aortic endothelial cell apoptosis under different hypoxia conditions. Methods: 24 SPF grade wistar rats were randomly divided into 3 groups, 8 rats in each group: normal oxygen control group (NC group), intermittent hypoxia group (IH group) and persistent hypoxia group (PH group). The expression of endoplasmic reticulum stress signal molecule BIPHOPOP-caspase12 in endothelial cells of aortic arch was detected by immunohistochemistry and Tunel method was used to detect the apoptosis of endothelial cells in aortic arch, and human umbilical vein endothelial cells were cultured in vitro. They were randomly divided into three groups: normoxic control group (NC1), normoxic control inhibitor group (NC1 TUDCA), intermittent hypoxia group (IH1), intermittent hypoxia inhibitor group (IH1 TUDCA), persistent hypoxia (4H) group (PH1), persistent hypoxia (4H) inhibitor group (Ph 1 TUDCA), and persistent hypoxia (4H inhibitor group). In 12 H group, H 2 group was treated with continuous hypoxia, and 12 H inhibitor group was treated with H 2 TUDCA group. Apoptosis was detected by flow cytometry and real-time PCR was used to detect the mRNA expression of endoplasmic reticulum stress factor BIPCHOPP caspase12 in all groups. Results: In vivo and in vitro, the results showed that the apoptosis rate of endothelial cells in intermittent hypoxia group and continuous hypoxia group increased P0.05, while the expression of endoplasmic reticulum stress factor BIPCHOPP caspase12 was significantly higher than that in normal control group, among which, at the same time, the expression of endoplasmic reticulum stress factor BIPCHOPP caspase12 was significantly increased. The apoptosis rate of endothelial cells and the expression of endoplasmic reticulum stress factor in intermittent hypoxia group were significantly higher than those in continuous hypoxia group. After adding endoplasmic reticulum stress inhibitor (TUDCA), the apoptosis rate of endothelial cells and the expression of endoplasmic reticulum stress factor BIPP-CHOP-caspase12 decreased significantly (P 0.05). Conclusion: Hypoxia (especially intermittent hypoxia) can induce apoptosis of aortic endothelial cells. Endoplasmic reticulum stress pathway may be an important pathway in the process of hypoxia induced apoptosis of aortic cells.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563.9

【共引文献】