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TRPA1激动剂桂皮醛防止高糖介导的心肌细胞氧化应激损伤

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2018年第2期
页码:
136-140,145
栏目:
基础研究
出版日期:
2018-02-15

文章信息/Info

Title:
TRPA1 agonist cinnamaldehyde protects against high glucose-induced oxidative stress injury
作者:
王 芳; 王 丹; 侯霁芯; 万进东; 刘家欣; 阚竞宇; 王沛坚
（成都医学院第一附属医院心内科、四川省高校衰老与血管稳态重点实验室，四川 成都 610500）
Author(s):
WANG Fang;  WANG Dan;  HOU Ji-xin;  WAN Jin-dong;  LIU Jia-xin;  KAN Jing-yu;  WANG Pei-jian
(Department of Cardiology, First Affiliated Hospital, Chengdu Medical College & Key Laboratory of Aging and Vascular Homeostasis, Sichuan Provincial Universities, Chengdu 610500, Sichuan, China)
关键词:
桂皮醛; 瞬时受体电位通道A1亚型; 氧化应激; 心肌细胞; 糖尿病
Keywords:
cinnamaldehyde;  transient receptor potential cation channel;  subfamily A;  member 1;  oxidative stress;  cardiomyocytes;  diabetes
分类号:
R363
DOI:
-
文献标识码:
A
摘要:
目的 探讨桂皮醛对高糖介导的心肌细胞氧化应激损伤的作用及机制。方法 大鼠心肌细胞（H9C2）以高糖培养基（33 mmol/L D-葡萄糖）培养，以低糖培养基（5.5 mmol/L D-葡萄糖）作为对照。首先利用Western blot和免疫组织化学证实瞬时受体电位通道A1亚型（TRPA1）在H9C2心肌细胞中的表达；在此基础上分别利用线粒体超氧化物荧光探针（mitoSOX）及TUNEL细胞凋亡试剂盒观察桂皮醛对高糖环境下H9C2细胞线粒体活性氧(ROS)水平以及细胞凋亡的影响，以Western blot法观察Nrf2和TRPA1的表达；利用荧光定量PCR观察TRPA1及Nrf2 mRNA水平。结果 TRPA1在H9C2细胞中有表达，桂皮醛可显著抑制高糖介导线粒体ROS生成（P<0.01），并可减少心肌细胞的凋亡（P<0.01），而上述作用可被TRPA1的阻断剂HC 030031所阻断。桂皮醛可上调TRPA1、Nrf2的蛋白及mRNA表达（P<0.01），而通过HC 030031抑制TRPA1可显著减弱桂皮醛的上述作用（P<0.01）。结论 桂皮醛可防止高糖介导的心肌细胞氧化应激损伤，而该作用可能与其激活TRPA1，上调Nrf2有关。
Abstract:
AIM To explore the effect and mechanism of cinnamaldehyde on oxidative stress injury induced by high glucose. METHODS Rat myocardial cells (H9C2) were cultured with both high glucose medium (33 mmol/L D-glucose) and low glucose medium (5.5 mmol/L D- glucose-e.g., control group). Western blotting and immunohistochemistry were used to observe the expression of TRPA1 in H9C2 cells. Then, mitochondrial superooxide fluorescence probe (mito-SOX) and TUNEL apoptosis kits were used to observe the effects of cinnamaldehyde on ROS, cell apoptosis in H9C2 and the expressions of Nrf2 and TRPA1 were observed by Western blotting. TRPA1 and Nrf2 mRNA levels were detected by fluorescence quantitative PCR. RESULTS Western blotting and immunohistochemistry confirmed that TRPA1 was expressed in H9C2 cells. Cinnamaldehyde inhibited the ROS generation from mitochondria (P<0.01) and decreased the cells apoptosis (P<0.01) under high glucose condition and these effects were reversed by pretreatment with TRPA1 blocker HC 030031. Fluorescence quantitative PCR and Western blotting indicated that cinnamaldehyde up-regulated both the protein and mRNA levels of TRPA1 and Nrf2 (P<0.01) under high glucose condition and these effects were also blocked by HC 030031 (P<0.01). CONCLUSION Cinnamaldehyde prevents the oxidative stress injury induced by high glucose as a TRPA1 agonist and this effect is associated with Nrf2 activation.

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备注/Memo

备注/Memo:
收稿日期：2017-04-13.基金项目：国家自然科学基金项目资助(81400289)；四川省科技厅应用基础研究项目资助(2015JY0275)；四川省科技计划青年基金项目资助(2016JQ0032)；四川省卫生计生委科研项目资助(150047)；四川省教育厅自然科学重点项目资助（16ZA0277） 通讯作者：王沛坚，副教授，主要从事心血管及代谢性疾病的基础及临床研究 Email:wpjmed@aliyun.com 作者简介：王芳，住院医师，硕士 Email:273037430@qq.com
更新日期/Last Update: 1900-01-01