基于网络药理学和ApoE^(-/-)小鼠动脉粥样硬化模型探究血府逐瘀汤治疗冠心病的核心作用靶点及机制研究

Core Target and Mechanism of Xuefu Zhuyu Decoction in the Treatment of Coronary Heart Disease Based on Network Pharmacology and ApoE^(-/-) Mice Atherosclerosis Model

目的基于网络药理学探索血府逐瘀汤治疗冠心病的核心作用靶点及机制,然后制备ApoE^(-/-)小鼠动脉粥样硬化模型进行验证。方法筛选血府逐瘀汤药物活性成分、作用靶点及冠心病致病靶点,然后筛选血府逐瘀汤与冠心病的交集靶点并构建成分-靶点网络;使用Cytoscape软件分析核心靶点,使用GO功能和KEGG通路富集分析预测血府逐瘀汤与冠心病的交集靶点所包含的信号通路。选取5~6周龄雄性C57BL/6小鼠7只和ApoE^(-/-)小鼠30只,分为低脂对照组:C57BL/6小鼠7只,进食低脂饲料;高脂模型组:ApoE^(-/-)小鼠10只,进食高脂饲料;低剂量灌胃组:ApoE^(-/-)小鼠10只,进食高脂饲料,然后每10 g鼠重给予0.1 ml血府逐瘀汤药液(1 g/ml)灌胃;高剂量灌胃组:ApoE^(-/-)小鼠10只,进食高脂饲料,然后每10 g鼠重给予0.1 ml血府逐瘀汤药液(2 g/ml)灌胃。比较四组小鼠血脂指标,高脂模型组、低剂量灌胃组、高剂量灌胃组小鼠主动脉组织胶原纤维面积/斑块总面积、斑块总面积/血管腔面积及缺氧诱导因子1α(HIF-1α)、血管内皮生长因子受体2(VEGFR2)阳性面积/斑块总面积。结果GO功能富集分析结果显示,根据q值进行排序,前3个条目为磷代谢过程的正向调控(q=7.43E^(-48))、细胞内信号转导的正向调节(q=3.45E^(-47))、细胞运动的正向调节(q=5.60E^(-41))。KEGG通路富集分析结果显示,根据q值降序排列,HIF-1α信号通路位列第1。高脂模型组小鼠LDL、TC高于低脂对照组,HDL低于低脂对照组(P<0.05);低剂量灌胃组、高剂量灌胃组小鼠LDL、TC低于高脂模型组,HDL高于高脂模型组(P<0.05)。低剂量灌胃组和高剂量灌胃组小鼠主动脉组织胶原纤维面积/斑块总面积高于高脂模型组,斑块总面积/血管腔面积低于高脂模型组,高剂量灌胃组小鼠主动脉组织胶原纤维面积/斑块总面积高于低剂量灌胃组,斑块总面积/血管腔面积低于低剂量灌胃组(P<0.05)。低剂量灌胃组和高剂量灌胃组小鼠主动脉组织HIF-1α、VEGFR2阳性面积/斑块总面积低于高脂模型组,高剂量灌胃组小鼠主动脉组织HIF-1α、VEGFR2阳性面积/斑块总面积低于低剂量灌胃组(P<0.05)。结论HIF-1α信号通路是血府逐瘀汤治疗冠心病的核心作用靶点之一,血府逐瘀汤可以通过降低血脂、抑制HIF-1α信号通路而减小动脉粥样硬化斑块面积及维持斑块的稳定性,进而有效治疗冠心病。

Objective To explore the core target and mechanism of Xuefu Zhuyu decoction in the treatment of coronary heart disease based on network pharmacology and then verify them via the ApoE^(-/-)mice atherosclerosis model.Methods The active components,action targets of Xuefu Zhuyu decoction,and pathogenic targets of coronary heart disease were screened.Then drugs-disease junction targets were screened,the component target network was constructed.Core targets were analyzed using Cytoscape software,and GO function and KEGG pathway enrichment analysis were used to predict the signal pathways involved in drug-disease junction targets.Seven male C57BL/6 mice and 30 ApoE^(-/-)mice aged 5-6 weeks were selected and divided into the following groups:low-fat control group:seven C57BL/6 mice were fed with low-fat diet;high-fat model group:10 ApoE^(-/-)mice were fed with high-fat diet;low-dose gavage group:10 ApoE^(-/-)mice were fed with the high-fat diet,and then 0.1 ml of Xuefu Zhuyu decoction(1 g/ml)was given every 10 g of mice weight;high-dose gavage group:10 ApoE^(-/-)mice were fed with the high-fat diet,and then 0.1 ml of Xuefu Zhuyu decoction(2 g/ml)was given every 10 g of mice weight.The blood lipid indexes were compared in the four groups;collagen fiber area/total plaque area,total plaque area/vascular lumen area,hypoxia-inducible factor 1α(HIF-1α)and vascular endothelial growth factor receptor 2(VEGFR2)positive area/total plaque area of the aortic tissue were compared in the high-fat model group,low-dose gavage group and high-dose gavage group.Results GO function enrichment analysis results showed that the top three items were the positive regulation of phosphorus metabolism(q=7.43E^(-48)),the positive regulation of intracellular signal transduction(q=3.45E^(-47)),and the positive regulation of cell movement(q=5.60E^(-41))according to the q value.KEGG pathway enrichment analysis results show that the HIF-1αsignaling pathway ranked first according to the q value.LDL and TC of mice in the high-fat model group were higher than those in the low-fat control group,and HDL level was lower than those in the low-fat control group(P<0.05);LDL and TC levels of mice in low-dose gavage group and high-dose gavage group were lower than those in the high-fat model group,while HDL level was higher than those in high-fat model group(P<0.05).The collagen fiber area/total plaque area of the aortic tissue in the low-dose gavage group and high-dose gavage group was higher than that in the high-fat model group,total plaque area/vascular lumen area of the aortic tissue in the low-dose gavage group and high-dose gavage group was lower than that in the high-fat model group,and the collagen fiber area/total plaque area of the aortic tissue in the high-dose gavage group was higher than that in the low-dose gavage group,total plaque area/vascular lumen area of the aortic tissue in the high-dose gavage group was lower than that in the low-dose gavage group(P<0.05).The HIF-1αand VEGFR2 positive area/total plaque area of the aortic tissue in the low-dose gavage group and the high-dose gavage group were lower than those in the high-fat model group,and the HIF-1αand VEGFR2 positive area/total plaque area of the aortic tissue in the high-dose gavage group were lower than those in the low-dose gavage group(P<0.05).Conclusion HIF-1αsignal pathway is one of the core targets of Xuefu Zhuyu decoction in the treatment of coronary heart disease.Xuefu Zhuyu decoction can reduce the area of atherosclerotic plaque and maintain the stability of plaque by decreasing blood lipid and inhibiting the HIF-1αsignal pathway,and then effectively treat the coronary heart disease.