Xu Fan, Li Xiao, Liu Lanfang, Xiao Xu, Zhang Li, Zhang Shenglin, Lin Pingping, Wang Xiaojie, Wang Yongwei, Li Qingshan
Department of Oncology, Affiliated Hospital of Chengde Medical College, Chengde, Hebei 067000, P.R. China.
Department of Radiology, Affiliated Hospital of Chengde Medical College, Chengde, Hebei 067000, P.R. China.
Exp Ther Med. 2017 Sep;14(3):2216-2220. doi: 10.3892/etm.2017.4763. Epub 2017 Jul 10.
The protective effects and mechanisms of esculetin on doxorubicin (DOX)-induced injury of H9c2 cells were investigated. H9c2 cells were cultured and the logarithmic growth phase of the cells was divided into a control group, a DOX group and an esculetin + DOX group. Cell viability was detected by MTT assay. Annexin V-PI (AV-PI) double staining flow cytometry was carried out to detect cell apoptosis. Intracellular reactive oxygen species (ROS) were detected by flow cytometry. Transmission electron microscope (TEM) was used to evaluate cell ultrastructure. Cleaved caspase-3, cleaved PARP, Bcl-2, Bid and Bmi-1 proteins levels were investigated by western blot analysis. Bmi-1 siRNA was used to detect the role of Bmi-1 in the protective effects of esculetin against DOX-induced toxicity in H9c2 cells. The MTT and AV-PI double staining results showed that esculetin significantly increased H9c2 cell viability. Compared with the control group, the levels of cleaved caspase-3, cleaved PARP, Bid and ROS levels were significantly decreased, but the expression of Bcl-2 and Bmi-1 were significantly increased in the esculetin + DOX group. TEM showed that the cell structure of the mitochondria was protected by esculetin. The results of Bmi-1 siRNA showed that esculetin could protect DOX-induced cardiotoxicity by modulating Bmi-1 expression. Esculetin can protect DOX-induced cardiotoxicity and the effects may be attributable to modulation of Bmi-1 expression, provoking intracellular ROS accumulation, protecting the structure of mitochondria and reducing cell apoptosis.
研究了七叶亭对阿霉素（DOX）诱导的H9c2细胞损伤的保护作用及机制。培养H9c2细胞，将细胞对数生长期分为对照组、DOX组和七叶亭+DOX组。采用MTT法检测细胞活力。进行Annexin V-PI（AV-PI）双染流式细胞术检测细胞凋亡。采用流式细胞术检测细胞内活性氧（ROS）。用透射电子显微镜（TEM）评估细胞超微结构。通过蛋白质免疫印迹分析研究裂解的caspase-3、裂解的PARP、Bcl-2、Bid和Bmi-1蛋白水平。使用Bmi-1 siRNA检测Bmi-1在七叶亭对H9c2细胞DOX诱导毒性的保护作用中的作用。MTT和AV-PI双染结果表明，七叶亭显著提高了H9c2细胞活力。与对照组相比，七叶亭+DOX组中裂解的caspase-3、裂解的PARP、Bid水平和ROS水平显著降低，但Bcl-2和Bmi-1的表达显著增加。TEM显示七叶亭保护了线粒体的细胞结构。Bmi-1 siRNA的结果表明，七叶亭可通过调节Bmi-1表达来保护DOX诱导的心脏毒性。七叶亭可以保护DOX诱导的心脏毒性，其作用可能归因于调节Bmi-1表达、引发细胞内ROS积累、保护线粒体结构和减少细胞凋亡。
