西红花酸对阿霉素诱导大鼠心肌细胞损伤的影响  

Effects of Crocetin on Adriamycin-induced Cardiomyocyte Injury in Rats

作　　者：韩笑[1] 王攀 刘建勋[1] 陶夏莉 任钧国[1] 付建华[1] 许勇刚[1] HAN Xiao;WANG Pan;LIU Jianxun;TAO Xiali;REN Junguo;FU Jianhua;XU Yonggang(Institute of Basic Medical Sciences of Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing Key Laboratory of Pharmacology of Chinese Materia Medica,Beijing 100091,China;Beijing Reno Technology Co.,Ltd.,Beijing 100089,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所,北京市中药药理重点实验室,北京100091 [2]北京瑞诺众德科技有限公司,北京100089

出　　处：《中国中医药信息杂志》2021年第10期64-69,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家科技重大专项-重大新药创制(2017ZX09301018)。

摘　　要：目的观察西红花酸对阿霉素诱导的大鼠心肌细胞损伤的影响,从线粒体角度探讨其作用机制。方法培养H9c2大鼠心肌细胞,阿霉素诱导建立心肌细胞损伤模型。细胞分为正常组、模型组(阿霉素10μmol/L)、西红花酸高剂量组(阿霉素10μmol/L+西红花酸20μmol/L)、西红花酸低剂量组(阿霉素10μmol/L+西红花酸10μmol/L),给药6 h后CCK-8法检测细胞存活率,DCFH-DA荧光探针检测细胞内活性氧(ROS)水平,JC-1荧光探针检测线粒体膜电位,流式细胞术检测细胞凋亡率,Western blot检测Caspase-3、Bcl-2、Bax蛋白表达,免疫荧光检测Caspase-8蛋白表达。结果与正常组比较,模型组H9c2细胞存活率降低(P<0.01),细胞内ROS水平升高(P<0.001),线粒体膜电位降低(P<0.001),细胞凋亡率升高(P<0.001),cleaved Caspase-3蛋白表达升高,cleaved Caspase-3/pro Caspase-3比值升高(P<0.01),Bcl-2蛋白表达降低,Bax蛋白表达升高,Bcl-2/Bax比值显著降低(P<0.001),Caspase-8蛋白表达明显升高(P<0.001);与模型组比较,西红花酸高、低剂量组H9c2细胞存活率显著升高(P<0.01,P<0.05),ROS水平显著降低(P<0.001),线粒体膜电位升高(P<0.001),细胞凋亡率降低(P<0.01),cleaved Caspase-3蛋白表达降低,cleaved Caspase-3/pro Caspase-3比值降低(P<0.001,P<0.05),Caspase-8蛋白表达显著降低(P<0.001),西红花酸高剂量组Bcl-2蛋白表达升高,Bax蛋白表达降低,Bcl-2/Bax比值显著升高(P<0.01)。结论西红花酸可保护阿霉素致心肌细胞损伤,其机制可能与保护线粒体、抑制细胞凋亡相关。Objective To observe the effects of crocetin on adriamycin-induced cardiomyocyte injury in rats;To discuss its mechanism from the perspective of mitochondria.Methods H9c2(rat cardiomyocytes)were cultured and stimulated by adriamycin to make cardiomyocyte damage.The experiment was divided into normal group,model group(adriamycin 10μmol/L),crocetin high-dosage group(adriamycin 10μmol/L+crocetin 20μmol/L),and crocetin low-dosage group(adriamycin 10μmol/L+crocetin 10μmol/L).After 6 hours of treatment,CCK-8 method was used to detect the cell survival rate;DCFH-DA was used to detect intracelluar ROS level;JC-1 fluorescent probe was used to detect mitochondrial membrane potential;flow cytometry was used to detect apoptosis rate;Western blot was used to detect the protein expressions of Caspase-3,Bcl-2 and Bax;immunofluorescence was used to detect the expression of Caspase-8.Results Compared with the normal group,the survival rate of the model group decreased(P<0.01),the level of intracelluar ROS increased(P<0.001),the mitochondrial membrane potential decreased(P<0.001),the apoptosis rate increased(P<0.001),the expressions of cleaved Caspase-3 and cleaved Caspase-3/pro Caspase-3 increased(P<0.01),the expression of Bcl-2 decreased,the expression of Bax increased,the ratio of Bcl-2/Bax decreased significantly(P<0.001),and the expression of Caspase-8 increased(P<0.001).Compared with the model group,crocetin high-and low-dosage groups significantly increased cell survival rate(P<0.01,P<0.05),decreased intracellular ROS level(P<0.001),increased mitochondrial membrane potential(P<0.001),and decreased apoptosis rate(P<0.01).The protein expression of cleaved Caspase-3 decreased and cleaved Caspase-3/pro Caspase-3 significantly decreased(P<0.001,P<0.05),and Caspase-8 protein expression was significantly reduced(P<0.001).The expressions of Bcl-2 increased and Bax decreased in crocetin high-dosage group,and the ratio of Bcl-2/Bax significantly increased(P<0.01).Conclusion Crocetin can protect adriamycin-induced cardiomyocyte i

关 键 词：西红花酸 阿霉素 线粒体 凋亡 心肌细胞 大鼠 

分 类 号：R285.5[医药卫生—中药学]