摘要
目的:通过构建细胞和动物模型,从不同层面探讨没食子酸对脓毒性心肌病的保护作用。方法:进行体外实验:用脂多糖(2μg/mL)诱导H9c2细胞建立损伤模型,将培养的H9c2细胞分为四组:空白对照组、模型组、没食子酸对照组和没食子酸治疗组;采用DAPI染色法检测细胞核凝聚;采用非荧光探针DCFH-DA评估细胞活性氧ROS堆积。进行体内实验:采用盲肠结扎穿孔术(CLP)构建脓毒症小鼠模型,将雄性小鼠随机分为四组:CLP组、sham组、没食子酸对照组和没食子酸治疗组,于造模后24 h处死小鼠,开胸摘取心脏,采用DHE染色法检测心肌组织ROS水平。结果:体外实验:与对照组相比,模型组心肌细胞死亡率和ROS水平均升高;与模型组相比,没食子酸治疗组的上述指标均有明显下降,差异具有统计学意义。体内实验:CLP组心肌组织ROS水平高于sham组,而没食子酸治疗组心肌组织ROS水平低于CLP组。结论:没食子酸对脂多糖诱导的H9c2细胞损伤和CLP诱导的脓毒性心肌损伤具有一定的保护作用,其机制可能与抑制细胞死亡、减少ROS产生有关。
Objective:To investigate the protective effect of gallic acid on septic cardiomyopathy from different levels by constructing cell and animal models.Methods:In vitro experiment:H9c2 cells were induced by lipopolysaccharide(2μg/mL)to establish damage model.Cultured H9c2 cells were divided into four groups:blank control group,model group,gallic acid control group and gallic acid treatment group.Nuclear agglutination was detected by DAPI staining.Non-fluorescent probe DCFH-DA was used to evaluate the accumulation of reactive oxygen species(ROS).In vivo experiments:Sepsis mouse model was constructed by cecal ligation and perforation(CLP),and male mice were randomly divided into four groups:CLP group,sham group,gallic acid control group and gallic acid treatment group.The mice were sacrificed 24 h after modeling,and the hearts were removed by chest opening.The ROS levels of myocardial tissue were detected by DHE staining.Results:In vitro experiment:Compared with the control group,the myocardial cell mortality and ROS levels were increased in the model group.Compared with the model group,the above indexes in the gallic acid treatment group were significantly decreased,and the difference was statistically significant.In vivo experiment:The ROS level of myocardium in CLP group was higher than that in sham group,while the ROS level of myocardium in gallic acid treatment group was lower than that in CLP group.Conclusion:Gallic acid can protect H9c2 cell damage induced by lipopolysaccharide and septic myocardial damage induced by CLP.The mechanism may be related to inhibiting cell death and reducing ROS production.
作者
王鹤晶
周雨桦
邓悠然
罗安妮
何杉
李欣芳
陈维倩
WANG Hejing;ZHOU Yuhua;DENG Youran;LUO Anni;HE Shan;LI Xinfang;CHEN Weiqian(Soochow Medical College,Soochow University,Suzhou Jiangsu 215123;Department of Cardiac and Vascular Surgery,The First Affiliated Hospital of Soochow University,Suzhou Jiangsu 215006;Institute of Cardiovascular Disease,Soochow Medical College,Soochow University,Suzhou Jiangsu 215006)
基金
苏州大学校级大学生创新创业训练计划项目(2021xj067)。
