全反式维A酸抑制细胞内凋亡信号和改善心肌缺血再灌注损伤的研究  

All-trans retinoid acid protects against myocardial ischemia reperfusion injury via inhibiting apoptosis signaling

作　　者：赵潇然 陈岩[2] 朱劲舟[1] 杨克[1] 陈秋静[1] 朱政斌[1] 金清清 张凤如[3] 

机构地区：[1]上海交通大学医学院附属瑞金医院心血管内科,上海200025 [2]民航上海医院内科,上海200336 [3]上海交通大学医学院附属瑞金医院老年病科,上海200025

出　　处：《内科理论与实践》2014年第3期189-192,共4页Journal of Internal Medicine Concepts & Practice

基　　金：国家自然科学基金青年科学基金项目(项目编号:81000050)

摘　　要：目的:探讨全反式维A酸(ATRA)对心肌缺血再灌注(IR)损伤的影响和可能机制。方法:用ATRA对大鼠心肌母细胞株H9c2进行预处理,之后通过缺氧-再氧干预建立体外心肌IR模型。干预后的H9c2细胞用水溶性四唑盐试剂(WST-1)法检测细胞存活率,膜联蛋白Ⅴ流式细胞术检测细胞凋亡情况,蛋白质印迹检测与细胞凋亡相关的活化胱天蛋白酶-3、B淋巴细胞瘤-2基因(Bcl-2)的表达以及促分裂原活化蛋白激酶(MAPKs)家族信号p38 MAPK(p38)、c-Jun氨基末端激酶(JNK)和细胞外信号调节蛋白激酶(ERK)的磷酸化表达变化。结果 :IR后ATRA0.01μmol/L和1μmol/L处理组H9c2细胞的存活率升高(P<0.01)。与对照组相比,ATRA 1μmol/L处理组明显抑制了H9c2细胞由IR引起的细胞凋亡(5.8%±0.5%比2.6%±0.2%,P<0.01),使IR后H9c2细胞的Bcl-2表达升高(0.78±0.01比1.08±0.02,P<0.01),胱天蛋白酶-3表达降低(1.24±0.03比0.97±0.04,P<0.01),p38(1.28±0.18比0.81±0.08)、JNK(1.21±0.01比0.93±0.03)、ERK 1/2(1.19±0.02比0.93±0.02)信号的磷酸化表达降低(均P<0.01)。结论:ATRA提高了IR后H9c2细胞的存活率,抑制了IR后H9c2细胞的凋亡,这种作用可能与MAPKs信号表达下调有关。Objective To investigate the effect of all-trans retinoid acid （ATRA） on myocardial ischemia-reperfusion （IR） injury and explore the related mechanism. Methods H9c2 myocardial cells were pretreated with ATRA for 24 h and then subjected to hypoxia/re-oxygenation injury （6-hour hypoxia and 1-hour re-oxygenation）. ATRA＇s effect on cell viability in H9c2 ceils was measured using a water soluble tetrazolium salt-1 （WST-1） assay, and cell apoptosis was quantified by annexin V-fluorescein isothiocyanate （FITC） based flow cytometry. Apoptosis-related activated caspase-3, Bcl-2 and mitogen-activated protein kinase （MAPKs） signaling were measured by Western blotting. Results WST-1 assay revealed greater cell viability in ATRA 0.01 Ixmol/L and 1 μmollL treated group （P〈0.01）. Flow-cytometry analysis demonstrated ATRA＇s （1 μmol/L） anti-apoptotic effect on IR in H9c2 cells （5.8%±0.5% vs 2.6%±0.2%, P〈0.01）. Western blotting revealed that compared with controls, ATRA pretreatment increased Bcl-2 expression （0.78±0.01 vs 1.08±0.02, P〈0.01）, reduced activated caspase-3 （1.24±0.03 vs 0.97±0.04, P〈0.01） and phosphorylation of p38 （1.28±0.18 vs 0.81± 0.08）, c-Jun Nterminal kinases （JNK） （1.21±0.01 vs 0.93±0.03）, and extracellular signal-regulated kinases （ERK） （1.19± 0.02 vs 0.93±0.02） （all P〈0.01）. Conclusions Our results showed that ATRA effectively increased H9c2 cell survival rate and reduced H9c2 cell apoptosis after I/R in vitro. This anti-apoptotic effect might be associated with down-regulation of MAPKs signaling.

关 键 词：全反式维A酸 心肌缺血再灌注损伤 细胞凋亡 促分裂原活化蛋白激酶家族 

分 类 号：R541[医药卫生—心血管疾病]