黄酮醇类化合物对心肌细胞凋亡过程caspase-3、Bcl-2、Bax蛋白表达影响  

Effect of Flavonols on Caspase-3,Bcl-2 and Bax Expression in Cardiomyocytes Apoptosis

作　　者：王丽伟[1] 刘凤芝[1] 陆景坤[1] 刘有昌[2] 杨宝峰[3] 

机构地区：[1]哈尔滨医科大学药学院药化与天然药化教研室,哈尔滨150081 [2]哈尔滨医科大学药学院物理化学教研室,哈尔滨150081 [3]哈尔滨医科大学药学院药理学教研室,哈尔滨150081

出　　处：《中国药学杂志》2007年第10期749-753,共5页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(30371684);黑龙江省自然科学基金资助项目(D0301)

摘　　要：目的研究槲皮素、杨梅素、桑色素和芦丁对大鼠心肌细胞凋亡的保护作用。方法体外培养Wistar乳鼠心肌细胞,建立心肌细胞凋亡模型;随机分组:正常组,损伤组和用药组。用药组分别加入30μmol.L-1的4种黄酮醇类化合物,孵育1h后,除正常组之外,加入H2O2诱导细胞凋亡。加入不同浓度的药物后,采用MTT法观察细胞存活情况,来进行药物有效浓度的筛选。采用TUNEL、流式细胞仪法检测心肌细胞凋亡情况,免疫组化法检测caspase-3蛋白、Bcl-2、Bax蛋白的表达情况。结果MTT结果显示,4种黄酮醇类在5和30μmol.L-1时对细胞没有毒性。4种黄酮醇类化合物与损伤组比较,槲皮素、杨梅素、桑色素的心肌细胞凋亡率降低、Bcl-2/Bax比值增大、抑制caspase-3蛋白的活化。结论槲皮素、杨梅素和桑色素对心肌细胞凋亡有保护作用,可以通过抑制caspase-3蛋白的活化、上调Bcl-2蛋白的表达或是降低Bax的表达来发挥作用,槲皮素表现出较强的抗凋亡活性。OBJECTIVE To study the protective effect of flavonols （ quercetin, myricetin, morin and rutin） on apoptosis of neonatal rat cardiomyocytes induced by H2 O2. METHODS The neonatal rat cardiomyocytes were cultured for 48 h in vitro, H2 02 （ 100μmol·L^-1 ） was used to build an oxidative stress-induced injury model. Cardiomyocytes were randomly divided into several groups： normal, model, cultured with different flavonols （30 μmol ·L^-1）. After being cultured for 1 h, apoptosis of Neonatal rat cardiomyocytes were induced by H2 O2 except normal group. The cytotoxicity of different flavonols were determined by MTT assay. Apoptosis rates was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling（TUNEL） and flow cytometry（FCM） . Im- munohistochemistry was used for the detection of caspase-3, Bcl-2 and Bax protein. RESULTS Each flavonol showed no cytotoxicity at 5, 30 μmol · L^-1 by MTT assay. Compared with model group, the apoptosis rates of quercetin, myricetin and morin were de- creased,increased the ratio of Bcl-2/Bax, and inhibited the activiation of caspase-3. CONCLUSION Quercetin, myricetin, morin have protective effects on apoptosis of cardiomyocytes. The protective mechanism could be related to its ability to inhibit the activation of caspase-3 protein , up-regulate the expression of Bcl-2, down-regulate the expression of Bax, quercetin shows a strong anti-apoptotic activity.

关 键 词：心肌细胞 黄酮醇类 凋亡 

分 类 号：R965[医药卫生—药理学]