■中药(TCM)桂枝甘草汤(GGD)的作用机制尚不清楚。
■本研究探讨了GGD抗心肌肥厚的机制。
■进行网络药理学分析以确定GGD的潜在靶标。体内实验,将C57BL/6J小鼠分为Con,去氧肾上腺素(PE,10mg/kg/d),2-氯腺苷(CADO,腺苷的稳定类似物,2mg/kg/d),ggd(5.4g/kg/d)和ggd(5.4g/kg/d)+CGS15943(一种非选择性腺苷受体拮抗剂,4mg/kg/d)。体外实验,原代新生大鼠心肌细胞(NRCM)分为Con,PE(100µM),CADO(5µM),GGD(10-5g/mL)和GGD(10-5g/mL)+CGS15943(5µM)。超声波,H&E和Masson染色,进行肥大基因的表达和细胞表面积以验证GGD的疗效。通过实时聚合酶链反应(PCR)检测腺苷受体(ADORs)的表达,蛋白质印迹和免疫荧光分析。
■网络药理学在GGD的核心靶标中确定了ADORs。体外实验表明,GGD减弱了PE诱导的表面积增加(EC50为5.484×10-6g/mL)。体内数据显示GGD减弱了PE诱导的心室壁增厚。体外和体内数据表明,GGD减轻了PE诱导的肥大基因表达(例如,ANP,BNP和MYH7/MYH6),A1AR过度表达和A2aAR下调。此外,CADO的作用类似于GGD,而CGS15943消除了GGD的大部分影响。
■我们的研究结果表明了GGD抑制心脏肥大的机制,强调ADORs的调节是HF的潜在治疗策略。
UNASSIGNED: The mechanisms of Traditional Chinese Medicine (TCM) Guizhi-Gancao Decoction (GGD) remain unknown.
UNASSIGNED: This study explores the mechanisms of GGD against cardiac hypertrophy.
UNASSIGNED: Network pharmacology analysis was carried out to identify the potential targets of GGD. In vivo experiments, C57BL/6J mice were divided into Con,
phenylephrine (PE, 10 mg/kg/d), 2-chloroadenosine (CADO, the stable analogue of adenosine, 2 mg/kg/d), GGD (5.4 g/kg/d) and GGD (5.4 g/kg/d) + CGS15943 (a nonselective adenosine receptor antagonist, 4 mg/kg/d). In vitro experiments, primary neonatal rat cardiomyocytes (NRCM) were divided into Con, PE (100 µM), CADO (5 µM), GGD (10-5 g/mL) and GGD (10-5 g/mL) + CGS15943 (5 µM). Ultrasound, H&E and Masson staining, hypertrophic genes expression and cell surface area were conducted to verify the GGD efficacy. Adenosine receptors (ADORs) expression were tested via real-time polymerase chain reaction (PCR), western blotting and immunofluorescence analysis.
UNASSIGNED: Network pharmacology identified ADORs among those of the core targets of GGD. In vitro experiments demonstrated that GGD attenuated PE-induced increased surface area (with an EC50 of 5.484 × 10-6 g/mL). In vivo data shown that GGD attenuated PE-induced ventricular wall thickening. In vitro and in vivo data indicated that GGD alleviated PE-induced hypertrophic gene expression (e.g., ANP, BNP and MYH7/MYH6), A1AR over-expression and A2aAR down-expression. Moreover, CADO exerts effects similar to GGD, whereas CGS15943 eliminated most effects of GGD.
UNASSIGNED: Our findings suggest the mechanism by which GGD inhibits cardiac hypertrophy, highlighting regulation of ADORs as a potential therapeutic strategy for HF.