摘要
目的·研究选择性α_1肾上腺素能受体激动剂去氧肾上腺素(phenylephrine, Phe)及拮抗剂哌唑嗪(prazosin,Pra)对阿霉素(doxorubicin,DOX)诱导心肌细胞凋亡的影响。方法·将大鼠H9C2心肌细胞分为4组;除对照组外,其他3组均加入1.8μmol/L DOX;DOX组仅给予DOX,激动剂组同时加入0.1 mmol/L Phe,拮抗剂组同时加入10μmol/L Pra。细胞培养24 h后,采用流式细胞术及TUNEL染色检测细胞凋亡;Western blotting检测凋亡效应物cleaved caspase 3的表达;实时定量PCR检测Bcl-2家族抗凋亡基因及促凋亡基因在m RNA水平的表达;CCK-8检测细胞活力。结果·Phe抑制DOX诱导的心肌细胞凋亡,cleaved caspase 3表达量降低;Pra促进DOX诱导的心肌细胞凋亡,cleaved caspase 3表达量升高。与DOX组比较,激动剂组Bcl-2、Bcl-w在m RNA水平的表达升高,Bax、Bad在m RNA水平的表达降低;而拮抗剂组Bcl-2、Bcl-w的表达降低,Bax、Bad的表达升高。给药24 h后,激动剂组心肌细胞活力较DOX组增高,拮抗剂组细胞活力与DOX组无明显差异。结论·DOX诱导的心肌细胞凋亡受α_1肾上腺素能信号通路调节。
Objective · To investigate the effects of α 1-adrenergic receptor agonist phenylephrine (Phe) and antagonist prazosin (Pra) on cardiomyocyte apoptosis induced by doxorubicin (DOX). Methods · H9C2 cardiomyocytes were divided into 4 groups. Except for the control group incubated with medium alone, all other groups were treated with 1.8 μmol/L DOX. For agonist group and antagonist group, 0.1 mmol/L Phe and 10 μmol/L Pra were added respectively in DOX-treated cells. After culture for 24 h, flow cytometry and TUNEL assay were performed to detect the apoptosis rate. Western blotting was used to detect the expression of cleaved caspase 3. Real-time PCR was used to detect the expression of anti-and pro-apoptotic Bcl-2 family genes. CCK-8 assay was used to detect the cell viability. Results · The DOX-induced apoptosis was inhibited by Phe with decreased apoptosis rate of H9C2 and decreased expression of cleaved caspase 3, but promoted by Pra. Increased expression of Bcl-2 and Bcl-w and decreased expression of Bax and Bad at mRNA levels were found in agonist group in comparison with the cells treated with DOX alone;while decreased expression of Bcl-2 and Bcl-w and increased expression of Bax and Bad were found in antagonist group. The cell viability after 24 h of treatment with agonist was higher than cells treated with DOX alone, but no signifiant difference was found in cell viability between antagonist group and DOX group. Conclusion ·α 1-Adrenergic signaling pathway may be involved in endogenous myocardial protection in the process of cardiomyocyte apoptosis induced by DOX.
作者
叶玉娇
李卓妍
王青洁
李文娟
陈笋
YE Yu-jiao;LI Zhuo-yan;WANG Qing-jie;LI Wen-juan;CHEN Sun(Department of Pediatric Cardiology, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China)
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2019年第3期233-238,共6页
Journal of Shanghai Jiao tong University:Medical Science
基金
国家自然科学基金(81741066
81700226)~~
