环氧二十碳三烯酸对内皮细胞内质网应激的作用及机制-内科学(心血管)专业论文.docxVIP

华 华 中 科 技 大 学 硕 士 学 位 论 文 PAGE 2 PAGE 2 细胞活性分别为 0.575±0.041 和 0.898±0.12，这表明，11,12-EET 能明显降低 2h-11 细胞的凋亡百分数并增加细胞活性（P0.05）；在 HUVECs 中，TM+DMSO 组和 TM+14,15-EET 组的凋亡百分数分别为：（41.673±3.756）%和（22.970±1.732）%， 细胞活性分别为 1.131±0.028 和 1.594±0.094，这表明，14,15-EET 能明显降低 HUVECs 的凋亡百分数并增加细胞活性（P0.05）。 3、G 蛋白抑制剂 GDP-β-s、Gs 蛋白抑制剂 NF-449、腺苷酸环化酶（AC）抑 制剂 SQ22536、蛋白激酶 A（PKA）抑制剂 H-89 和蛋白激酶 G（PKG）抑制剂 KT-5823 与 TM+EETs 组相比，Grp78 水平明显增加（P0.05）。而 EGFR、MEK、PI3K 和 PPAR-γ 抑制剂与 TM+EETs 组相比，Grp78 水平无明显改变，无统计学差异（P0.05）。 结论：1、外源性的EETs可以改善由衣霉素诱导的内皮细胞的内质网应激和细胞凋亡， 并增加细胞的活性。 2、外源性的EETs改善由衣霉素诱导的内皮细胞的内质网应激的机制可能是通过 G蛋白偶联受体，进而激活PKA和PKG。 3、外源性的 EETs 改善由衣霉素诱导的内皮细胞的内质网应激的机制可能不通 过激活 PI3K、EGFR、MEK 和 PPAR-γ。 关键词：环氧二十碳三烯酸；2h-11 细胞；脐静脉内皮细胞；内质网应激；细胞凋亡； The effects and mechanisms of epoxyeicosatrienoic acids on endoplasmic reticulum stress in endothelial cells Abstract Objective: To examine the effects of arachidonic cytochrome P450 epoxygenase metabolite epoxyeicosatrienoic acids (EETs) on endoplasmic reticulm (ER) stress in endothelial cells and potential molecular mechanisms. Methods: 1, Human umbilical vein endothelial cells (HUVECs) and 2h-11 cells were treated with 10μM tunicamycin (TM) to induce ER stress and then detected ER stress-related proteins, such as p-PERK, Grp78 and Xbp-1 using Western Blot. 2, Exogenous EETs (8,9-EET, 11,12-EET and 14,15-EET) were added. ER stress related proteins such as p-PERK, Grp78 and Xbp-1 were detected using Western Blots and apoptosis of endothelial cells was assessed by flow cytometry. Cells viability was determined with cells counting assay-8. 3, Related pathway inhibitors (Inhibitors of G protein, Gs protein, adenylate cyclase, protein kinase A, protein kinase G, PI3K, EGFR, MEK and PPAR-γ) were added. ER stress related proteins Grp78 was detected using Western Blots. Results: 1, TM treatment significantly increased the expression of ER stress related proteins p-PERK, Grp78 and Xbp-1 with maximum at 4h in 2h-11 cells and 8h in HUVECs (P0.05). 2, Exogenous EETs atten