基于网络药理学探讨养血培本健脑膏对治疗阿尔茨海默病的作用机制

Study on the mechanism of blood-nourishing,root-fostering and brain-strengthening cream in the treatment of Alzheimer's disease based on network pharmacology

目的探讨养血培本健脑膏对治疗阿尔茨海默病(AD)的作用机制。方法利用TCMSP数据库结合文本挖掘方式,提取养血培本健脑膏的化合物组成、化合物靶点关系,利用OMIM和GeneCards网站提取“Alzheimer′s disease”“mild cognitive impairment(MCI)”“dementia”的疾病靶点,R语言V3.6.3绘制Venn图以获得交集靶点,相关信息导入Cytoscape V3.7.2以获得化合物-靶点-疾病网络;依据交集靶点,STRING网站获得蛋白互作网络,R语言V3.6.3进行GO、KEGG富集分析,并对交集靶点数最多的10种化合物进行分子对接研究;双侧海马注射β淀粉样蛋白(Aβ)25~35建立AD动物模型,并用养血培本健脑膏灌胃治疗28 d,免疫组织化学染色观察大脑海马Aβ、Bcl-2蛋白表达。结果养血培本健脑膏含有97种化合物,与疾病的交集靶点149个,靶点间存在相互作用,部分化合物可调节β淀粉样蛋白前体(APP)、丝裂原活化蛋白激酶(MAPK)1、白细胞介素(IL)-6活性;GO、KEGG富集分析发现,神经递质受体通路、G蛋白耦联胺受体、流体剪切应力与老年性硬化、糖尿病并发症AGE-RAGE通路等可能为养血培本健脑膏治疗AD的机制;动物实验显示养血培本健脑膏可改善海马CA1区神经元Aβ、Bcl-2的表达(P<0.05)。结论养血培本健脑膏治疗AD具有多靶点、多途径和多通路的特点,其治疗机制可能与MAPK1、IL-6活性有关,且与其降低海马CA1区神经元Aβ表达、提高Bcl-2表达有关。

Objective To explore the mechanism of blood-nourishing, root-fostering and brain-strengthening cream in the treatment of Alzheimer’s disease(AD). Methods TCMSP database and text mining were used to extract the compound composition and compound target relationship of blood-nourishing, root-fostering and brain-strengthening cream. OMIM and GeneCards websites were used to extract the disease targets of "Alzheimer’s disease", "mild cognitive improvement(MCI)" and "dementia". Venn map was drawn in R language V3.6.3 to obtain the intersection targets, and relevant information was imported into Cytoscape V3.7.2 to obtain the compound-target-disease network. According to the intersection targets, the STRING website obtained the protein interaction network, the R language V3.6.3 carried on the GO, KEGG enrichment analysis, and the molecular docking research to the 10 compounds with the most intersection targets was carried on. The animal model of AD was established by injecting Aβ25~35 into the bilateral hippocampus, and the rats were treated with blood-nourishing, root-fostering and brain-strengthening cream orally for 28 days. The expressions of Aβ and Bcl-2 protein in the hippocampus were observed by immunohistochemistry.Results There were 97 kinds of compounds in blood-nourishing, root-fostering and brain-strengthening cream, 149 intersection targets with diseases, and some of them could regulate the activities of APP, MAPK1 and IL-6. The enrichment analysis of GO and KEGG showed that neurotransmitter receptor pathway, G protein-coupled amine receptor, fluid shear stress and atherosclerosis and AGE-RAGE signaling pathway in diabetic complications might be the mechanism of prescription in the treatment of AD. Animal experiments showed that the prescription could improve the expression of Aβ and Bcl-2 in hippocampal CA1 neurons(P<0.05).Conclusions Blood-nourishing, root-fostering and brain-strengthening cream had the characteristics of multi-target, multi-channel and multi-pathway in the treatment of AD. Its mechanism might be related to MAPK1 and IL-6 activity, and it might be related to the decrease of Aβ expression and the increase of Bcl-2 expression in hippocampal CA1 neurons.