摘要
目的:研究甘木通总黄酮对表柔比星心肌毒性的抑制作用及机制。方法:从初生SD大鼠分离培养原代心肌细胞,以1.0μmol/L表柔比星建立心肌毒性模型,采用表面活性剂-超声协同方法提取甘木通中总黄酮,设3个药物浓度组,在损伤模型的基础上分别加入终浓度为25、50、100μg/m L的甘木通总黄酮,阳性对照组加入10μmol/L右丙亚胺,各组均作用24 h,采用CCK-8检测细胞存活率,Hoechst33258染色观察细胞的凋亡形态,JC-1荧光探针检测线粒体膜电位变化,Western blot检测凋亡相关信号蛋白Bax、Bcl-2及cleaved Caspase-3的表达量,采用比色法测定超氧化物歧化酶(SOD)活性以及丙二醛(MDA)的生成量。结果:与模型组比较,甘木通总黄酮各剂量均可显著提高心肌细胞的存活率,降低表柔比星对线粒体的损伤,并可通过降低Bax及上调Bcl-2的表达量抑制cleaved Caspase-3的生成,进而抑制细胞的凋亡(P<0.05);同时能提高SOD的活性,抑制脂质过氧化反应从而降低MDA的生成(P<0.05)。结论:甘木通总黄酮可通过抑制脂质过氧化反应干预细胞凋亡的线粒体通路从而降低表柔比星的心肌毒性。
Objective：To study the protective effect and mechanism of total flavones of Clematis filamentosa（TFC）mitigating cardiotoxicity which induced by epirubicin（Epi）.Methods：The myocardial cells separated from neonate SD rats was induced to apoptosis with 1.0 μmol/L epirubicin to establish cardiotoxicity model.Three experimental groups were treated with 25,50,100 μg/mL TFC respectively for 24 h,while the dexrazoxane group was treated with 10 μmol/L dexrazoxane for the same period.Viability of myocardial cells was measured by CCK-8 kit.Apoptotic morphology was observed by fluorescent inverted microscope after Hoechst33258 staining.Mitochondrial transmembrane potential was determined by JC-1 kit.Expression levels of Bax,Bcl-2 and cleaved Caspase-3 were determined by Western blot.The concentration of SOD and MDA was assayed by colorimetric kit to investigate oxidative damage.Results：Compared with model group,viability of myocardial cells was significantly higher and the apoptosis was lower in TFC treament groups.Mitochondrion was also protected by TFC from the toxicity of epirubicin.The activity of SOD was enhanced,and the production of MDA was reduced significantly which treated by TFC （P 〈 0.05）,and the apoptosis was attenuated by causing higher expression of Bax,Bcl-2 and inhibited the production of cleaved Caspase-3 which treated with TFC.Conclusion：TFC can inhibit mitochondria-dependent apoptosis pathway via attenuating lipid peroxidation and thus mitigated cardiac toxicity of epirubicin.
出处
《中药材》
CAS
北大核心
2017年第2期436-440,共5页
Journal of Chinese Medicinal Materials
基金
广东省中医药局中医药强省项目(20141197
20161035)
关键词
甘木通
黄酮
表柔比星
右丙亚胺
心肌毒性
Clematis filamentosa Dunn
Flavones
Epirubicin
Dexrazoxane
Cardiac toxicity
