摘要
目的:研究参附注射液对慢性心力衰竭模型大鼠心肌内源代谢产物的影响,探讨益气温阳法疗效机制。方法:将SD大鼠随机分为正常组、模型组,模型组通过多点皮下注射异丙肾上腺素法制备慢性心力衰竭心阳虚证大鼠模型,造模7 d后喂养14 d,成模大鼠随机分为模型组、参附注射液组(6.0 mL·kg^(-1))、曲美他嗪组(10.0 mg·kg^(-1)),进行药物干预15 d。正常组、模型组注射等剂量生理盐水,完成后取材。进行心脏彩超检测,苏木素-伊红(HE)染色观察组织病理形态学,酶联免疫吸附测定法(ELISA)检测血清N末端脑钠肽前体(NT-proBNP)含量,透射电镜(TEM)观察心肌细胞线粒体形态结构变化,利用超高效液相色谱-质谱联用法(UHPLC-Q-MS)进行代谢轮廓分析,结合正交偏最小二乘法-判别分析(OPLS-DA)等方法筛选差异代谢物并进行鉴别,再运用MetaboAnalyst数据库进行进一步的筛选,通过通路富集分析得到相关生物学途径。受试者工作特征(ROC)曲线评价各潜在生物标志物对心肌损伤的诊断价值及药效的评估价值。结果:彩超结果表明,参附注射液可以明显改善模型大鼠心功能指标(P<0.05);HE染色结果显示,参附注射液可有效改善模型大鼠的心肌组织结构紊乱、炎性细胞浸润等病理现象;ELISA结果显示,参附注射液有效调节模型大鼠的血清NT-proBNP水平;TEM观察发现,参附注射液可有效恢复线粒体形态结构;代谢组学结果显示各组心肌样本的代谢表型差异明显,参附注射液组可显著回调9种差异代谢物,涉及3条代谢通路,包括丙酮酸代谢,组氨酸代谢,柠檬酸(TCA)循环等能量代谢途径;ROC曲线分析结果显示,所有代谢物的曲线下面积(AUC)值均介于0.75~1.0,表明差异代谢物对心肌损伤具有较高的诊断准确性,其表达水平变化可作为潜在的药效评估标志物。结论:参附注射液通过改善能量代谢重构显著改善慢性心力衰竭心阳虚证模型大鼠的心功能、心肌及线粒体结构损伤,其生物学机制涉及能量代谢等相关途径,益气温阳法是治疗慢性心力衰竭心阳虚证的重要方法之一。
Objective:To examine the influences of Shenfu injection on the endogenous metabolic byproducts in the myocardium of the rat model exhibiting chronic heart failure,thus deciphering the therapeutic mechanism of the Qi-reinforcing and Yang-warming method.Methods:SD rats were randomly allocated into a control group and a modeling group.Chronic heart failure with heart-Yang deficiency syndrome in rats was modeled by multi-point subcutaneous injection of isoproterenol,and the rats were fed for 14 days after modeling.The successfully modeled rats were randomized into model,Shenfu injection(6.0 mL·kg^(-1)),and trimetazidine(10 mg·kg^(-1))groups and treated with corresponding agents for 15 days.The control group and the model group were injected with equal doses of normal saline,and the samples were collected after the intervention was completed.Cardiac color ultrasound was performed.Hematoxylin-eosin(HE)staining was used to observe histopathological morphology,and the serum level of N-terminal pro-brain natriuretic peptide(NT-proBNP)was assessed by enzyme-linked immunosorbent assay(ELISA).The mitochondrial morphological and structural changes of cardiomyocytes were observed by transmission electron microscopy,and the metabolic profiling was carried out by ultra high performance liquid chromatography-quantitative exactive-mass spectrometry(UHPLC-QE-MS).Differential metabolites were screened and identified by orthogonal partial least squares-discriminant analysis(OPLS-DA)and other methods,and then the MetaboAnalyst database was used for further screening.The relevant biological pathways were obtained through pathway enrichment analysis.The receiver operating characteristic(ROC)curve was established to evaluate the diagnostic value of each potential biomarker for myocardial injury and the evaluation value for drug efficacy.Results:The results of color ultrasound showed that Shenfu Injection improved the cardiac function indexes of model rats(P<0.05).The results of HE staining showed that Shenfu injection effectively alleviated the pathological phenomena such as myocardial tissue structure disorder and inflammatory cell infiltration in model rats.The results of ELISA showed that Shenfu injection effectively regulated the serum NT-proBNP level in the model rats.Transmission electron microscopy(TEM)showed that Shenfu injection effectively restored the mitochondrial morphological structure.The results of metabolomics showed that the metabolic phenotypes of myocardial samples presented markedly differences between groups.Nine differential metabolites could be significantly reversed in the Shenfu injection group,involving three metabolic pathways:pyruvate metabolism,histidine metabolism,and citric acid cycle(TCA cycle).The results of ROC analysis showed that the area under the curve(AUC)values of all metabolites were between 0.75 and 1.0,indicating that the differential metabolites had high diagnostic accuracy for myocardial injury,and the changes in their expression levels could be used as potential markers for efficacy evaluation.Conclusion:Shenfu injection significantly alleviated the damage of cardiac function,myocardium,and mitochondrial structure in the rat model of chronic heart failure with heart-Yang deficiency syndrome by ameliorating energy metabolism remodeling.Reinforcing Qi and warming Yang is a key method for treating chronic heart failure with heart-Yang deficiency syndrome.
作者
宁欣玥
赵震宇
张梦娜
郭阳
向芝佳
廉坤
胡志希
李琳
NING Xinyue;ZHAO Zhenyu;ZHANG Mengna;GUO Yang;XIANG Zhijia;LIAN Kun;HU Zhixi;LI Lin(Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Provincial Key Laboratory of Traditional Chinese Mdeicine Diagnostics,Hunan University of Chinese Medicine,Changsha 410208,China;The Domestic First-class Discipline Construction Project of Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)
出处
《中国实验方剂学杂志》
北大核心
2026年第5期178-186,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82305092,82274412)
湖南中医药大学科研基金“揭榜挂帅”专项(2022XJJB002)
湖南省青年科技创新人才项目(2024RC3199)
2024年湖南省大学生创新创业训练计划项目(S202410541015)。
关键词
慢性心力衰竭
能量代谢重构
中药干预
参附注射液
代谢组学
chronic heart failure
energy metabolic remodeling
traditional Chinese medicine intervention
Shenfu injection
metabolomics
