Song Miao-Yu, Zhu Cui-Ling, Li Yi-Zhuo, Li Xing-Yuan, Liu Gang, Li Xiao-Hui, Sun Yan-Qin, DU Ming-Yuan, Jiang Lei, Yue Chao-Chong
Heart Center/National Regional(Traditional Chinese Medicine)Cardiovascular Diagnosis and Treatment Center,the First Affiliated Hospital of Henan University of Chinese Medicine Zhengzhou 450000,China Henan University of Chinese Medicine Zhengzhou 450046,China.
Heart Center/National Regional(Traditional Chinese Medicine)Cardiovascular Diagnosis and Treatment Center,the First Affiliated Hospital of Henan University of Chinese Medicine Zhengzhou 450000,China.
Zhongguo Zhong Yao Za Zhi. 2025 Apr;50(8):2184-2192. doi: 10.19540/j.cnki.cjcmm.20241217.401.
This study aims to explore the protective effect of Chaihu Jia Longgu Muli Decoction on rats with heart failure after myocardial infarction, and to clarify its possible mechanisms, providing a new basis for basic research on the mechanism of classic Chinese medicinal formula-mediated inflammatory response in preventing and treating heart failure induced by apoptosis after myocardial infarction. A heart failure model after myocardial infarction was established in rats by coronary artery ligation. The rats were divided into sham group, model group, and low, medium, and high-dose groups of Chaihu Jia Longgu Muli Decoction, with 10 rats in each group. The low-dose, medium-dose, and high-dose groups of Chaihu Jia Longgu Muli Decoction were given 6.3, 12.6, and 25.2 g·kg~(-1) doses by gavage, respectively. The sham group and model group were given an equal volume of distilled water by gavage once daily for four consecutive weeks. Cardiac function was assessed using color Doppler echocardiography. Myocardial pathology was detected by hematoxylin-eosin(HE) staining, apoptosis was measured by TUNEL assay, and mitophagy was observed by transmission electron microscopy. The levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-1β, and N-terminal pro-B-type natriuretic peptide(NT-proBNP) in serum were detected by enzyme-linked immunosorbent assay(ELISA). The expression of apoptosis-related proteins B-cell lymphoma 2(Bcl-2), Bcl-2-associated X protein(Bax), and cleaved caspase-3 was detected by Western blot. Additionally, the expression of phosphorylated nuclear transcription factor-κB(NF-κB) p65(p-NF-κB p65)(upstream) and nuclear factor kappa B inhibitor alpha(IκBα)(downstream) in the NF-κB signaling pathway was assessed by Western blot. The results showed that compared with the sham group, left ventricular ejection fraction(LVEF) and left ventricular short axis shortening(LVFS) in the model group were significantly reduced, while left ventricular end diastolic diameter(LVEDD) and left ventricular end systolic diameter(LVESD) increased significantly. Myocardial tissue damage was severe, with widened intercellular spaces and disorganized cell arrangement. The apoptosis rate was increased, and mitochondria were enlarged with increased vacuoles. Levels of TNF-α, IL-1β, and NT-proBNP were elevated, indicating an obvious inflammatory response. The expression of pro-apoptotic factors Bax and cleaved caspase-3 increased, while the anti-apoptotic factor Bcl-2 decreased. The expression of p-NF-κB p65 was upregulated, and the expression of IκBα was downregulated. In contrast, the Chaihu Jia Longgu Muli Decoction groups showed significantly improved of LVEF, LVFS and decreased LVEDD, LVESD compared to the model group. Myocardial tissue damage was alleviated, and intercellular spaces were reduced. The apoptosis rate decreased, mitochondrial volume decreased, and the levels of TNF-α, IL-1β, and NT-proBNP were lower. The expression of pro-apoptotic factors Bax and cleaved caspase-3 decreased, while the expression of the anti-apoptotic factor Bcl-2 increased. Additionally, the expression of p-NF-κB p65 decreased, while IκBα expression increased. In summary, this experimental study shows that Chaihu Jia Longgu Muli Decoction can reduce the inflammatory response and apoptosis rate in rats with heart failure after myocardial infarction, which may be related to the regulation of the IκBα/NF-κB signaling pathway.
本研究旨在探讨柴胡加龙骨牡蛎汤对心肌梗死后心力衰竭大鼠的保护作用,并阐明其可能机制,为经典中药方剂介导的炎症反应防治心肌梗死后凋亡所致心力衰竭的机制基础研究提供新依据。采用冠状动脉结扎法建立大鼠心肌梗死后心力衰竭模型。将大鼠分为假手术组、模型组以及柴胡加龙骨牡蛎汤低、中、高剂量组,每组10只。柴胡加龙骨牡蛎汤低、中、高剂量组分别按6.3、12.6、25.2 g·kg⁻¹剂量灌胃给药。假手术组和模型组每日灌胃等体积蒸馏水,连续四周。采用彩色多普勒超声心动图评估心功能。通过苏木精-伊红(HE)染色检测心肌病理,采用TUNEL法检测凋亡,通过透射电子显微镜观察线粒体自噬。采用酶联免疫吸附测定(ELISA)法检测血清中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β和N末端B型利钠肽原(NT-proBNP)水平。采用蛋白质印迹法检测凋亡相关蛋白B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和裂解的半胱天冬酶-3的表达。此外,采用蛋白质印迹法评估核因子κB(NF-κB)信号通路中磷酸化核转录因子κB p65(p-NF-κB p65)(上游)和核因子κB抑制蛋白α(IκBα)(下游)的表达。结果显示,与假手术组相比,模型组左心室射血分数(LVEF)和左心室短轴缩短率(LVFS)显著降低,而左心室舒张末期内径(LVEDD)和左心室收缩末期内径(LVESD)显著增加。心肌组织损伤严重,细胞间隙增宽,细胞排列紊乱。凋亡率增加,线粒体肿大,空泡增多。TNF-α、IL-1β和NT-proBNP水平升高,表明存在明显的炎症反应。促凋亡因子Bax和裂解的半胱天冬酶-3表达增加,而抗凋亡因子Bcl-2表达降低。p-NF-κB p65表达上调,IκBα表达下调。相反,与模型组相比,柴胡加龙骨牡蛎汤各剂量组LVEF、LVFS显著改善,LVEDD、LVESD降低。心肌组织损伤减轻,细胞间隙减小。凋亡率降低,线粒体体积减小,TNF-α、IL-1β和NT-proBNP水平降低。促凋亡因子Bax和裂解的半胱天冬酶-3表达降低,而抗凋亡因子Bcl-2表达增加。此外,p-NF-κB p65表达降低,而IκBα表达增加。综上所述,本实验研究表明柴胡加龙骨牡蛎汤可降低心肌梗死后心力衰竭大鼠的炎症反应和凋亡率,这可能与调节IκBα/NF-κB信号通路有关。