二参真武汤对慢性心力衰竭大鼠炎性反应及凋亡途径的影响

Therapeutic effects of Ershen Zhenwu Decoction on inflammatory response and apoptotic pathways in chronic heart failure rats

关键词:  慢性心力衰竭  二参真武汤  网络药理学  分子机制

Key words:  chronic heart failure  Ershen Zhenwu Decoction  network pharmacology  molecular mechanism

摘要:

[目的] 通过网络药理学结合实验验证，探析二参真武汤治疗慢性心力衰竭（CHF）的分子机制。[方法] 通过TCMSP和Uniprot数据库筛选二参真武汤的成分及靶点，使用Genecards和OMIM数据库确定CHF的相关靶点；利用STRING平台获取交集靶点的蛋白相互作用数据，借助Cytoscape 3.8.2构建核心靶点网络；利用David数据库进行基因本体富集分析（GO富集分析）和京都基因与基因组百科全书富集分析（KEGG富集分析），并绘制药物-成分-靶点-通路图。将35只大鼠随机分为正常组、模型组及二参真武汤低、高剂量组、对照组，使用阿霉素腹腔注射建立CHF大鼠模型。通过检测血清氨基末端B型钠尿肽前体（NT-ProBNP）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）含量及苏木精-伊红（HE）染色观察心肌组织变化，并通过蛋白免疫印迹（Western blot）、实时荧光定量聚合酶链式反应（qPCR）法验证相关凋亡靶点的表达水平。[结果] 筛选得到85个药物活性成分、174个药物靶点和118个药物-疾病交集靶点；关键核心靶点为丝氨酸和苏氨酸激酶（AKT1）、B细胞淋巴瘤/白血病-2（Bcl-2）、基质金属蛋白酶-9（MMP9）、肿瘤坏死因子（TNF）、半胱氨酸天冬氨酸特异性蛋白酶3（CASP3）和IL-6等；GO分析涵盖810个条目；KEGG分析显示肿瘤坏死因子（TNF）、程序性细胞死亡（Apoptosis）、磷脂酰肌醇-3-激酶（PI3K）-蛋白激酶B（Akt）和丝裂原活化蛋白激酶（MAPK）等信号通路与交集靶点密切相关；药物-成分-靶点网络显示，儿茶素、芍药苷和丹参酮ⅡA与交集靶点高度关联。动物实验结果显示，与正常组比较，模型组人氨基末端B型钠尿肽前体（NT-proBNP）、IL-6、TNF-α、半胱氨酸天冬氨酸特异性蛋白水解酶（Caspase）-3和Bcl-2相关X蛋白（Bax）表达升高，Bcl-2表达下降（P<0.05），心肌组织炎性浸润增加；与模型组比较，二参真武汤组NT-proBNP、IL-6、TNF-α、Caspase-3和Bax表达下降，Bcl-2表达上升（P<0.05），心肌组织炎性浸润得到改善。[结论] 二参真武汤对CHF大鼠心肌具有保护作用，其作用机制可能与调控炎性反应和抑制细胞凋亡相关。

Abstract:

[Objective] To investigate the molecular mechanisms of Ershen Zhenwu Decoction（EZD） in treating chronic heart failure（CHF） through network pharmacology and experimental validation. [Methods] Active components and targets of EZD were screened using TCMSP and UniProt databases，while CHF-related targets were identified from GeneCards and OMIM databases. Protein-protein interaction networks of intersecting targets were constructed using STRING and visualized with Cytoscape 3.8.2. GO and KEGG enrichment analyses were performed using DAVID database. Thirty-five rats were randomly divided into normal，model，low-dose EZD，high-dose EZD，and control groups. The CHF model was established by intraperitoneal injection of doxorubicin. Serum levels of NT-proBNP，IL-6， and TNF-α were measured，and myocardial histopathology was examined by HE staining. The expression of apoptosis-related proteins was detected by Western blot and qRT-PCR. [Results] The study identified 85 active components，174 drug targets，and 118 drug-disease intersecting targets. Core targets included AKT1，Bcl-2，MMP9，TNF，CASP3，and IL-6. GO analysis revealed 810 enriched terms，while KEGG analysis showed significant associations with TNF，apoptosis，PI3K-Akt，and MAPK signaling pathways. The component-target network demonstrated strong associations between catechin，paeoniflorin，tanshinoneⅡA and the intersecting targets. Compared with normal group，model group showed elevated NT-proBNP，IL-6，TNF-α，Caspase-3 and Bax（P<0.05），decreased Bcl-2（P<0.05），and increased inflammatory infiltration. EZD treatment reversed these changes（P<0.05） and improved myocardial pathology. [Conclusion] EZD exerts cardioprotective effects in CHF rats，potentially through modulating inflammatory responses and inhibiting apoptosis.