摘要
该研究探讨木香烃内酯(costunolide,Cos)对阿霉素(doxorubicin,DOX)诱导心肌细胞损伤的保护作用及机制。在细胞实验中,将H9C2心肌细胞分为正常组、模型组、低剂量组、中剂量组、高剂量组和Rapamycin组,以DOX建立损伤模型。EDU检测细胞增殖活力、流式细胞术检测细胞凋亡率、免疫荧光染色检测LC3表达情况。在动物实验中,以SD大鼠为对象,将其随机分成5组:Ctrl组、Model组、L-Cos组、M-Cos组和H-Cos组,每组10只。彩色多普勒超声检测大鼠心脏功能。HE和Masson染色检测心肌组织病理变化。蛋白免疫印迹检测并分析与凋亡、自噬过程密切相关的蛋白表达水平,以揭示所研究生物体系中的潜在分子机制。电镜下观察心肌细胞线粒体内自噬体数目。(1)在细胞实验中,与正常组相比,模型组H9C2细胞的EDU阳性率显著降低(P<0.05);细胞凋亡率、LC3荧光强度显著增加(P<0.05)。与模型组相比,低剂量组、中剂量组和高剂量组H9C2细胞的EDU阳性率显著增加(P<0.05);细胞凋亡率、LC3荧光强度显著降低,呈剂量依赖性(P<0.05)。(2)在动物实验中,与Ctrl组相比,Model组大鼠的LVESD、LVEDD、p53、Bax和Bnip3蛋白表达水平,LC3-Ⅱ/LC3-Ⅰ值,心肌细胞线粒体自噬体数目显著增加(P<0.05);LVFS、LVEF、Bcl-2、P62蛋白表达水平显著降低(P<0.05)。与Model组相比,L-Cos组、M-Cos组和H-Cos组大鼠的LVESD、LVEDD、p53、Bax和Bnip3蛋白表达水平,LC3-Ⅱ/LC3-Ⅰ值,心肌细胞线粒体自噬体数目显著降低(P<0.05);LVFS和LVEF,Bcl-2、P62蛋白表达水平显著增加,呈剂量依赖性(P<0.05)。Cos能够阻断细胞凋亡信号通路、减缓心肌细胞线粒体过度自噬进程、增强细胞代谢活力以及减轻心肌纤维化病变程度,最终改善心脏功能,有效减轻DOX引发的心肌毒性损伤。
This study was to investigate the protective effect and mechanism of Cos(costunolide)on DOX(doxorubicin)-induced cardiomyocyte injury.In cell experiments,H9C2 cardiomyocytes were divided into normal group,model group,low-dose group,medium-dose group and high-dose group,and DOX was used to establish injury model.EDU was used to detect cell proliferation activity;flow cytometry was employed to precisely measure the apoptosis rate of the cells,while immunofluorescence staining was utilized to accurately detect the expression of LC3 protein,providing valuable insights into the cellular physiological state.In the animal experiment,a total of SD rats were randomly allocated into five distinct groups:Ctrl group,Model group,L-Cos group,M-Cos group,and H-Cos group,with ten rats in each group.The cardiac function parameters of these rats were measured.Additionally,HE staining and Masson’s trichrome staining were employed to examine the pathological alterations in the myocardial tissue.Western blot was meticulously employed to detect and analyze the expression levels of proteins that were closely associated with apoptosis and autophagy processes,aiming to uncover the underlying molecular mechanisms in the studied biological system.The number of autophagosomes in mitochondria of cardiomyocytes was observed under electron microscope.(1)In the cellular experiment,the proportion of EDU-positive H9C2 cells in the model group was notably decreased compared with that in the normal group(P<0.05).The rates of apoptosis and the fluorescence intensity of LC3 experienced a significant elevation(P<0.05).Compared with the model group,the EDU positive rate of H9C2 cells in the low-dose group,middle-dose group and high-dose group was significantly increased(P<0.05).The apoptosis rate and LC3 fluorescence intensity were significantly decreased in a dose-dependent manner(P<0.05).(2)In animal experiments,compared with the Ctrl group,LVESD,LVEDD,p53,Bax,and Bnip3 protein expression,LC3-II/LC3-I ratio,and the count of mitochondrial autophagosomes within cardiomyocytes were notably higher in the Model group(P<0.05).LVFS and LVEF,along with the protein expression levels of Bcl-2 and P62,were significantly reduced(P<0.05).Compared with the Model group,LVESD,LVEDD,p53,Bax,and Bnip3 protein expression,LC3-II/LC3-I ratio,and the number of mitochondrial autophagosomes in cardiomyocytes were significantly decreased in the L-Cos group,M-Cos group,and H-Cos group(P<0.05).LVFS and LVEF,Bcl-2 and P62 protein expression levels were significantly increased in a dose-dependent manner(P<0.05).Cos can inhibit the process of excessive mitophagy in cardiomyocytes,block the apoptotic signaling pathway,enhance the metabolic activity of cells and reduce the degree of myocardial fibrosis,and ultimately improve cardiac function and effectively resist DOX-induced myocardial toxic injury.
作者
吴玉
罗智
胡春华
WU Yu;LUO Zhi;HU Chunhua(Department of Cardiovascular Medicine,Mianyang People’s Hospital,Mianyang 621000,China;Department of Obstetrics,Mianyang People’s Hospital,Mianyang 621000,China)
出处
《中国细胞生物学学报》
2025年第11期2855-2866,共12页
Chinese Journal of Cell Biology
关键词
木香烃内酯
阿霉素
心肌细胞损伤
凋亡
自噬
costunolide
doxorubicin
myocardial cell injury
apoptosis
autophagy
