咖啡酸抗H2O2 诱导的人脐静脉内皮细胞凋亡及机制探讨

中国临床药理学与治疗学 ›› 2007, Vol. 12 ›› Issue (12): 1367-1375.

咖啡酸抗H₂O₂ 诱导的人脐静脉内皮细胞凋亡及机制探讨

熊丹, 严奉祥, 欧和生

南华大学药物药理研究所,衡阳 421001,湖南

收稿日期:2007-07-27 发布日期:2020-11-10
通讯作者: 严奉祥,男,教授,研究生导师,留美学者,研究方向:分子药理学和药物开发。Tel:0734-8281408 E-mail:fengxyan@sina.com
作者简介:熊舟,女,在读硕士生,研究方向:分子药理学。Tel:0734-8281408 E-mail:wjzzp520@163.com
基金资助:
湖南省卫生厅基金资助项目(204062);湖南省教育厅重点基金资助项目(03A040);国家教育部重点基金资助项目(204095);国家十五攻关项目(02EFN214300457)

Effects of caffeic acid on apoptosis of HUVEC-12 induced by H₂O₂ and the mechanism involved

XIONG Dan, YAN Feng-xiang, OU He-sheng

Institute of Pharmacology and Pharmacy, Nanhua University, Hengyang 421001, Hunan, China

Received:2007-07-27 Published:2020-11-10

摘要/Abstract

摘要： 目的: 用H₂O₂ 诱导人脐静脉内皮细胞株凋亡建立氧化应激模型,观察咖啡酸对H₂O₂ 诱导的人脐静脉内皮细胞株(HUVEC-12)凋亡的保护作用并初步探讨其机制。方法: 分别用不同浓度的咖啡酸和H₂O₂ 处理HUVEC-12,通过四甲基偶氮哇盐还原法(MTT法)检测细胞的活力,流式细胞术测定内皮细胞凋亡率,用吖叮橙溴化乙吱荧光染色(AO-EB染色)观察细胞凋亡形态,Western blotting检测NF-kB、P53和Bcl-2的表达。结果: 用 0.5 mmol/L H₂O₂和不同浓度咖啡酸共同孵育24 h,MTT提示咖啡酸能明显减轻H₂O₂ 对HUVEC-12的损伤,提高细胞的存活率,流式细胞检测术才是示咖啡酸能使H₂O₂ 诱导的 HUVE-12凋亡率由21.9%±2.1%降至3.8%± 1.1 %。AO-EB染色提示咖啡酸能降低H₂O₂ 诱导的细胞凋亡,使凋亡细胞减少;同时NF-kB和P53表达上调,Bcl-2表达下调。结论: 不同浓度的咖啡酸对H₂O₂ 引起的HUVEC-12损伤有保护作用,并呈现一定的浓度依赖性和时效性,其机制可能与调节NF KB、P53和Bcl-2蛋白的表达有关。

关键词: 咖啡酸, 人脐静脉内皮细胞, 凋亡, NF-kB

Abstract: AIM: To investigate the effects of caffeic acid on apoptosis of HUVEC-12 induced by H₂O₂, and to explore the related phaimacological mechanism. METHODS: The (cells were incubated with H₂O₂ (0. 5 mmol/L) and caffeic acid (1, 10, 100 µmol/L). The viability of HUVEC-12 was measured by MIT assay at different concentrations or times. Flow cytomet analysis was uesd to determine the rate of cell apoptosis. AO/EB dyeing was used to detect the morphological changes of cells. Westem blotting was used to detect the NF-kB, P53 or Bcl-2 protein expression. RESULTS: HUVEC-12 was pretreat ed with 1, 10, 100 µmol/L caffeic acid for 30 minutes followed by 0.5 mmol/L H₂O₂, the rate of cell survival reduced in dose-dependent manner. Results of flow cytoment demonstrated that the apoptotic rate of cells decreased from 21. 9 % ± 2. I % to 3. 8 % ± I. I %. Spontaneous apoptosis was significantly increased in the cells incubated with caffeic acid, meanwhile, the NF-kB protein was upregulated, Bcl-2 protein was down-re lated, ai1d P53 protein was up-regulated. CONCLUSION: Caffeic acid could resist the H₂O₂-induced cell apoptosis, with the possible molecular mechanism of regulating the NF-kB, P53 and BCL-2 expression.

Key words: caffeic acid, HUVEC-12, apoptosis, NF–κB

中图分类号:

R965.2

熊丹, 严奉祥, 欧和生. 咖啡酸抗H₂O₂ 诱导的人脐静脉内皮细胞凋亡及机制探讨[J]. 中国临床药理学与治疗学, 2007, 12(12): 1367-1375.

XIONG Dan, YAN Feng-xiang, OU He-sheng. Effects of caffeic acid on apoptosis of HUVEC-12 induced by H₂O₂ and the mechanism involved[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2007, 12(12): 1367-1375.

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咖啡酸抗H₂O₂ 诱导的人脐静脉内皮细胞凋亡及机制探讨

Effects of caffeic acid on apoptosis of HUVEC-12 induced by H₂O₂ and the mechanism involved

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