桃红四物汤活性成分的筛选及阿魏酸对心肌损伤保护作用的研究

目的：运用系统药理学方法筛选桃红四物汤中的活性成分并通过细胞实验研究其对心肌损伤的保护作用。方法：通过TCMSP数据库和Pharmmaper数据库获取桃红四物汤所有成分及其作用靶点,同时应用Genecards数据库获取心肌损伤相关疾病的潜在靶点。再利用Cytoscape构建并分析成分-疾病交集靶点网络图以及成分贡献度,筛选出桃红四物汤作用于心肌的主要活性成分。进一步应用CoCl2诱导H9c2心肌细胞建立缺氧损伤模型,检测所筛选出的活性成分对细胞活力、心肌酶水平、能量代谢的影响。明确桃红四物汤中的活性成分通过影响能量代谢发挥对心肌细胞损伤的保护作用。结果：筛选发现桃红四物汤包含73个活性成分,其对应340个作用靶点,而心肌损伤相关的疾病靶点为1 544个;根据成分-疾病交集靶点等网络图,筛选出槲皮素、阿魏酸和芍药苷是桃红四物汤中抗心肌损伤的主要活性成分。进一步实验发现阿魏酸能够显著升高CoCl2诱导的H9c2心肌细胞活力（P<0.05）,显著降低CoCl2刺激后心肌酶中肌酸激酶（CK）、肌酸激酶同工酶（CK-MB）水平以及乳酸脱氢酶（LDH）含量（P<0.01）。阿魏酸能够抑制CoCl2刺激后乳酸（LD）水平的升高,以及三磷酸腺苷（ATP）水平的降低（P<0.01）。 结论：本研究筛选发现桃红四物汤中的阿魏酸是抗心肌损伤的主要活性成分,进一步研究证明阿魏酸可通过抑制心肌损伤后的心肌酶水平和改善能量代谢障碍发挥保护心肌的作用。

Study on the screening of active constituents of Taohong Siwu Decoction and the protective effect of ferulic acid on myocardial injury

AIM: To screen the active constituents in Taohong Siwu Decoction(THSWD) by Systems Pharmacology and to study its protective effect on myocardial injury through cell experiments. METHODS: The TCMSP database and Pharmmaper database were used to retrieve the constituents and active constituent targets of THSWD. The Genecards database was used to retrieve the myocardial injury related targets gene. Then construct and analyze the constituents-disease intersection target network and the contribution value of constituents, and screen out the main active constituents of THSWD acting on the myocardium. Furthermore, CoCl2 was used to induce H9c2 to establish a hypoxic injury model, and the effects of the selected active constituents on cell viability were detected. The level of myocardial enzyme and energy metabolism were detected to investigate whether the active constituents in THSWD play a protective effect on myocardial injury by affecting energy metabolism. RESULTS: The screening found that THSWD contained 73 active constituents, which corresponded to 340 targets, while 1 544 disease targets related to myocardial injury. According to the network diagrams of constituents-disease intersection targets, quercetin, ferulic acid and paeoniflorin were screened out as the main active constituents of THSWD against myocardial injury. Further experiments found that ferulic acid can significantly increase the viability of H9c2 induced by CoCl2 (P<0.05), and significantly reduce the levels of CK, CK-MB and LDH after CoCl2 stimulation (P<0.01). In addition, ferulic acid can inhibit the increase of LD level and the decrease of ATP level after CoCl2 stimulation (P<0.01). CONCLUSION: Ferulic acid in THSWD is the main active constituent against myocardial injury and it can protect the myocardium by inhibiting myocardial enzyme levels and improving energy metabolism disorders.