摘要
目的:观察不同类型的黄酮类化合物红花黄色素A(查儿酮)、高良姜素(黄酮醇)、陈皮素(二氢黄酮)对H2O2诱导的心肌细胞凋亡及Bcl-2、Bax、Caspase-3蛋白表达的影响。方法:取Wistar大鼠乳鼠心肌细胞做原代培养,制备H2O2诱导的心肌细胞凋亡模型。MTT法检测3种黄酮类化合物适宜的实验浓度,TUNEL法、流式细胞术检测细胞凋亡率,免疫组化法测定Bcl-2、Bax、Caspase-3蛋白表达。结果:MTT法显示各组药物只有5μmol/L组与空白组相比差异无统计学意义(P<0.05)。仅30μmol/L组细胞活力显著高于模型组,随后实验选用30μmol/L作为试验浓度。TUNEL法检测细胞凋亡率,除红花黄色素A组外各药物组细胞凋亡率均显著小于模型组(P<0.05)。流式细胞术检测显示,各药物组均能降低细胞凋亡率和坏死率。免疫组化显示,Caspase-3蛋白阳性表达除红花黄色素A组(与空白对照组比较P>0.05)外各药物组均显著小于模型组(P<0.05)。Bcl-2/Bax比值除红花黄色素A组(与正常组比较P>0.05)外各药物组均显著高于模型组(P<0.05)。TUNEL法检测凋亡率、Caspase-3蛋白阳性表达、Bcl-2/Bax比值均显示陈皮素和高良姜素组间无统计学差异(P>0.05)。结论:陈皮素和高良姜素抑制100μmol/LH2O2诱导新生乳鼠心肌细胞凋亡的效果相当,可调节Bcl-2、Bax、Caspase-3蛋白表达,起到保护心肌细胞作用;而红花黄色素A对细胞坏死的影响可能比对细胞凋亡的作用更强。
To investigate the effect of different types of flavonoids, galangin （flavonol）, hesperetin （flavonone） and hydroxysafflor yellow A （HYSA, chalcone）, on cardiomyocytic injury induced by H202, and explore the possible signal pathways involved. METHODS： The cytotoxicity of different flavonoids was determined by MTTassay. Apoptosis rate was determined by flow cytometry （FCM） and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL）. lmmunohistochemistry was used for detection of Bcl-2, Bax and Caspase-3 protein. RESULTS： It showed that each flavonoid did not have noticeable cytotoxicity at a concentration of 5μmol/L by MTr assay. Flavonoids at concentrations of 5, 15 and 30 5μmol/L significantly increased cell viability compared to model group induced by H2O2 （100 5 μmol/L）. Flavonoids also increased apoptosis rote and neorobiosisrate determined by FCM compared to model group. Galangin and hesperetin significantly decreased the apoptotie rate determined by TUNEL and the expression of Caspase 3 and inereased the ratio of Bcl-2/Bax （ P 〈 0.05） compared with model group. CONCLUSION： The results suggest that galangin and hesperetin inhibit the apoptosis induced by H2O2 in neonatal rat cardiomyoeytes and that there was not noticeable difference between galangin and hesperefin in TUNEL test when H2O2 was used at a concentration of 100 5 μmol/L. They regulate the expression of Bcl-2, Bax and Caspase-3 proteins. HYSA may have stronger effect on neorobiesis than on apoptesis.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2007年第6期620-625,共6页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
国家自然科学基金项目(30371684)
