【作者】 王丽伟； 刘凤芝； 陆景坤； 刘有昌； 杨宝峰；

【Author】 Li-Wei Wang 1 , Feng-Zhi Liu 1, Jing-Kun Lu 1, you-chang Liu 2,Bao-Feng Yang3 (1. Department of Pharmacochemistry and Natural Pharmacochemistry, 2. Department of Physical chemistry, 3. Department of pharmacology,Harbin Medical University, Harbin 150081, China)

【机构】 哈尔滨医科大学药化与天然药化教研室； 哈尔滨医科大学物理化学教研室； 哈尔滨医科大学药理学教研室；

【摘要】 目的研究黄酮醇类化合物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。用药组分别加入四种黄酮醇类化合物,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。结果四种黄酮醇类化合物与损伤组比较,杨梅素槲皮素、桑色素的心肌细胞凋亡率降低、Caspase-3蛋白表达降低、Bcl-2/Bax比值增大。结论黄酮醇类化合物对心肌细胞凋亡有保护作用,可以通过上调Caspase-3蛋白、Bcl-2蛋白的表达或是降低Bax的表达来发挥作用。更多还原

【Abstract】 Object To study the protective effect of Flavonols(quercetin, myricetin, morin and rutin)on apoptosis of neonatal rat cardiomyocytes induced by H2O2. Methods The neonatal rat cardiomyocytes were cultured for 48 hours in vitro. H2O2 (100 μmol.L-1) was used to build an oxidative stress-induced injury model. Cardiomyocytes were randomly divided into several groups: control, model, cultured with different flavonols for 1 hour. The cytotoxicity of different flavonols were determined by MTT assay; Apoptosis rate was determined by flow cytometry(FCM) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling(TUNEL); Immunohistochemistry was used for detection of caspase-3, Bcl-2 and Bax protein. Results It showed that each flavonol did not have noticeable cytotoxicity in 5, 30 μmol.L-1 by MTT assay. Compared with model group, the apoptosis rate of quercetin, myricetin and morin decreased (P<0.05), caspase-3 activity attenuated and the ratio of Bcl-2/Bax increased (P<0.05). Conclusion Quercetin, myricetin, morin have protective effects on apoptosis of cardiomyocytes. The protective mechanism could be related to its ability to regulate the expression of Bcl-2, Bax and caspase-3 protein.更多还原