数字中医药（英文）2021，Vol.4Issue(3)：214-228,15.DOI:10.1016/j.dcmed.2021.09.006
平阳降压方治疗高血压的网络药理学研究及体内实验验证
Mechanism of Pingyang Jiangya Formula in treating hypertension based on network pharmacology and in vivo study
摘要
Abstract
Objective This study aimed to analyze the mechanism of action of the Pingyang Jiangya Formula (平阳降压方, PYJYF) in treating hypertension, based on network pharmacology, and to verify the subsequent predictions through animal experiments. Methods The active components and related target genes of PYJYF were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Bioinformatics Analysis Tool for Molecular Mechan-ism of Traditional Chinese Medicine (BATMAN-TCM), Encyclo-pedia of Traditional Chinese Medicine (ETCM), and DrugBank databases and available literature. The hypertension target genes were screened based on Therapeutic Target Database (TTD), GeneCards, Online Mendelian Inheritance in Man (OMIM), Uni-Prot, and relevant literature. The component-disease-target net-work intersection target genes were inputted into the STRING database, and the key target genes were selected according to the degree algorithm. Gene Ontology (GO) analysis and Kyoto Encyc-lopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed to explore the multitarget mechanism of action and molecular regulatory network of PYJYF in the treat-ment of hypertension. To verify this prediction, we used PYJYF to intervene in spontaneously hypertensive rats (SHRs) and Wistar–Kyoto rats (WKY) as normal control, and the noninvasive tail artery manometry method was used to measure systolic blood pressure (SBP) in the rat tail before PYJYF intervention. After drug intervention, the SBP of each group rats were measured and com-pared every week. Enzyme-linked immunosorbent assay (ELISA) was used to test plasma renin, angiotensin Ⅱ(Ang Ⅱ), and aldos-terone (Ald) levels, and hematoxylin-eosin (HE) staining was used to observe pathological damage to the renal vessels in each group of rats. Western blot and reverse transcription real-time quantitative PCR (RT-PCR) were used to detect the protein and mRNA expression levels of PI3K, AKT1, BAX, and Bcl-2, respect-ively.Results A total of 4123 hypertension targets were obtained from related databases. From the TCMSP and chemical databases, 78 active components of PYJYF and the corresponding 401 drug tar-gets were retrieved. Data analysis revealed that 208 drug targets directly interacted with the hypertension targets in PYJYF. The 10 targets most closely related to hypertension target proteins in PYJYF were directly retrieved from relevant databases. GO ana-lysis revealed that 10 direct target proteins were involved in all as-pects of the antihypertensive effects of PYJYF, as well as molecu-lar biological processes, such as the regulation of blood pressure, renin-angiotensin-aldosterone system (RAAS), angiotensin-me-diated ligand reactions, and biological stimulation of cardiomyo-cyte apoptosis. KEGG pathway enrichment analysis revealed that PYJYF directly affected 20 signaling pathways associated with hy-pertension. In animal experiments, PYJYF reduced the protein and mRNA levels of PI3K, Akt, and Bax and upregulated the ex-pression of the protein and mRNA levels of Bcl-2, reduced plasma renin, Ang Ⅱ, and Ald levels, improved the hyperactivity of RAAS, and significantly reduced SBP in SHRs. Conclusion PYJYF is effective for hypertension therapy that acts through multiple compounds and targets. The possible underly-ing molecular mechanism includes regulating the PI3K/Akt sig-naling pathway to suppress RAAS, increasing the ratio of Bcl-2/Bax proteins, and inhibiting apoptosis, thereby mediating the re-pair of renal and renal vascular damage caused by hypertension. These findings warrant further research for use in clinical set-tings.关键词
平阳降压方/高血压/网络药理学/PI3K/Akt信号通路/RASS/细胞凋亡/生物信息学Key words
Pingyang Jiangya Formula (平阳降压方,PYJYF)/Hypertension/Network pharmacology/PI3K/Akt signaling pathway/Renin-angiotensin-aldosterone system (RAAS)/Apoptosis/Bioinformatics引用本文复制引用
刘德果,李姿蓉,陈其华,王宇红,肖长江..平阳降压方治疗高血压的网络药理学研究及体内实验验证[J].数字中医药（英文）,2021,4(3):214-228,15.基金项目
We thank for the funding support from the National Natural Science Foundation of China(No.81874464),National Major New Drug Development Project(No.2019ZX09301-103),and Provincial Department of Graduate Research Innovation Project of Hunan(No.CX20190565). （No.81874464）
