[1]胡小春,刘浩宇,覃数,等.Exendin-4对人脐静脉内皮细胞内质网应激损伤的拮抗作用[J].陆军军医大学学报(原第三军医大学学报),2013,35(17):1827-1830.
 Hu Xiaochun,Liu Haoyu,Qin Shu,et al.Effect of exendin-4 on human umbilical vein endothelial cells with endoplasmic reticulum stress[J].J Amry Med Univ (J Third Mil Med Univ),2013,35(17):1827-1830.
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Exendin-4对人脐静脉内皮细胞内质网应激损伤的拮抗作用(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
35卷
期数:
2013年第17期
页码:
1827-1830
栏目:
论著
出版日期:
2013-09-15

文章信息/Info

Title:
Effect of exendin-4 on human umbilical vein endothelial cells with endoplasmic reticulum stress
作者:
胡小春刘浩宇覃数张冬颖刘剑
重庆医科大学附属第一医院心血管内科
Author(s):
Hu Xiaochun Liu Haoyu Qin Shu Zhang Dongying Liu Jian
Department of Cardiology, First Affiliated Hospital, Chongqing Medical University, Chongqing, 400016, China
关键词:
人脐静脉内皮细胞内质网应激钙联蛋白艾塞那肽细胞凋亡
Keywords:
human umbilical vein endothelial cells endoplasmic reticulum stress calnexin exendin-4 cell apoptosis
分类号:
R322.12; R329.28; R977.6
文献标志码:
A
摘要:
目的      研究exendin-4(Ex-4)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)内质网应激相关凋亡的拮抗作用。      方法        RPMI1640培养基培养HUVECs,待细胞长到培养瓶底部80%左右后,分4组干预细胞,每组8瓶,共32瓶。实验分组:①正常对照组(n=8):不做任何处理;②Ex-4组(n=8):(Ex-4,8 μg/mL,45 min);③EPBS组(n=8):(EPBS,100 μL/mL,30 min);④双药组(n=8):Ex-4预处理(8 μg/mL,45 min),后加EPBS(100 μL/mL,30 min)。免疫荧光法检测确认HUVECs上存在胰高血糖素样肽-1受体(glucagon-like peptide-1 receptor,GLP-1R);内源性过氧化氢酶抑制物(endogenous peroxidase blocking solution;EPBS)诱导HUVECs的凋亡,流式细胞仪检测细胞凋亡率变化;加入Ex-4预处理后,观察EPBS诱导HUVECs凋亡率的变化。Western blot检测细胞Calnexin、(C/EBP homologous protein,CHOP)和Bax表达情况,探讨内质网应激相关凋亡的机制。      结果        ① HUVECs上有GLP-1受体表达;②与正常对照组相比,EPBS组HUVECs的凋亡率显著增加[(52.27±3.66)%vs(7.25±0.62)%,P<0.01)];与EPBS组比较,Ex-4的预处理可明显降低EPBS诱导的HUVECs凋亡[(37.77±2.86)% vs(52.27±3.66)%,P<0.01)]。与正常对照组相比,EPBS能增加Calnexin [(1.76±0.16)vs(1.00±0.05),P<0.01]、CHOP [(4.58±0.32)vs(1.00±0.12),P<0.01]和Bax [(2.25±0.10)vs(1.00±0.06),P<0.01]的表达;与EPBS组比较,Ex-4的预处理可显著降低Calnexin [(0.42±0.05)vs(1.76±0.16),P<0.01]、CHOP [(0.64±0.10)vs(1.65±0.12),P<0.01]和Bax[(1.48±0.08)vs(2.25±0.10),P<0.01)]的表达。      结论        Ex-4对EPBS诱导的HUVECs凋亡有明显抑制作用,其抑制作用可能与EX-4减少内质网应激相关蛋白Calnexin、CHOP和Bax相关。
Abstract:
Objective       To study the antagonistic effect of exendin-4 (Ex-4) on human umbilical vein endothelial cells (HUVECs) with endoplasmic reticulum (ER) stress-induced cell apoptosis.       Methods        HUVECs were cultured with RPMI 1640 medium. When the HUVECs grew to around 80% at the bottom of culture flasks, the cells were divided into 4 groups (n=8) including: a control group (without any treatment), an Ex-4 group (Ex-4, 8 μg/ml for 45 min), an endogenous peroxidase blocking solution (EPBS) group (EPBS, 100 μL/mL for 30 min), and an Ex-4+EPBS group [Ex-4 pretreatment (8 μg/mL for 45 min) and EPBS treatment (100 μL/mL for 30 min)]. The glucagon-like peptide-1 receptor (GLP-1R) was verified by immunofluorescence method. The cell apoptosis was detected by flow cytometry. The expression of calnexin, C/EBP homologous protein (CHOP) and Bax were detected by Western blotting.       Results      GLP-1R existed in HUVECs. EPBS could significantly increase the apoptosis of HUVECs compared with the control group [(52.27±3.66)% vs (7.25±0.62)%, P<0.01)]. The rate of cell apoptosis in the Ex-4+EPBS group was significantly lower than that in the EPBS group [(37.77±2.86)% vs (52.27±3.66)%, P<0.01)]. Compared with the control group, EPBS group could increase the expression of calnexin [(1.76±0.16) vs (1.00±0.05), P<0.01], CHOP [(4.58±0.32)vs. (1.00±0.12), P<0.01] and Bax [(2.25±0.10) vs (1.00±0.06), P<0.01]. Compared with the EPBS group, Ex-4 group could significantly reduce the expression of calnexin [(0.42±0.05) vs (1.76±0.16), P<0.01], CHOP [(0.64±0.10)vs(1.65±0.12), P<0.01] and Bax [(1.48±0.08) vs (2.25±0.10), P<0.01)].       Conclusion        Ex-4 can inhibit EPBS-induced apoptosis in HUVECs through down-regulating the expression of ER stress-related proteins such as calnexin, CHOP and Bax.

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更新日期/Last Update: 2013-09-06