Study On The Effect And Mechanism Of Jiangya Kangxian Formula Conducted By Network Pharmacology

Objective:To build a new Chinese medicine compound Jiangya Kangxian Formula for the treatment of hypertensive myocardial fibrosis using network pharmacology and traditional Chinese medicine(TCM)theory,and to explore its therapeutic effects and mechanisms on hypertensive myocardial fibrosis in vivo and vitro.Method:(1)Data of Chinese herbs,ingredients,targets and diseases were obtained from TCMSP databases.Chinese herbs acting on targets of hypertension and myocardial fibrosis were selected using Cytoscape,then further screened according to TCM theory and combined together into a new Chinese medicine compound called Jiangya Kangxian Formula accoding the principles of group prescription.(2)High performance liquid chromatography-mass spectrometry(HPLC/MS)was used to construct the fingerprnnt of Jiangya Kangxian Formula,and the similarity analyses of each batch of drugs were carried out with the finger:print similarity evaluation system to establish the quality standard of Jiangya Kangxian Formula;(3)30 16-week-old Spontaneously hypertensive rats(SHRs)were taken as disease model and randomly divided into model group:spontaneous hypertensive rat group(SHR),Jiangya Kangxian Formula low dose group(KXL,2g/kg),Jiangya Kangxian Formula moderate dose group(KXM,4g/kg),Jiangya Kangxian Formula high dose group(KXH,8g/kg)and captopril positive control group(CAP,13.5mg/kg).Five rats with normal blood pressure(WKY)were taken as blank group.The rats were orally administered with drugs for consecutively 20 weeks and they were fasting for 12h before being put to death.Noninvasive tail voltmeter and Color doppler were used to detect blood pressure and cardiac function.Tissue staining and biochemical kit were used to observed heart histomorphology and serum biochemical indexes.(4)After analyzing active ingredients of Jiangya Kangxian Formula using HPLC/MS,the targets of the active ingredients were found by molecular docking,and the signaling pathways were analyzed through functional enrichment.The expression of proteins in related pathway were verified in vivo and vitro.(5)Gene chip was used to analyze differentially expressed genes in myocardium after administeration of Jiangya Kangxian Formula,and functional enrichment was analyzed.Ultimately,the signal pathways regulated by Jiangya Kangxian Formula were verified in vivo and vitro.Result:(1)It was confirmed that Jiangya Kangxian Decoction consist of Salvia miltiorrhiza,Panax notoginseng,Astragalus membranaceus,Millettia,and Cinnamomum camphora.(2)Fingerprint of Jiangya Kangxian Formula was constructed and the quality standard was establish by HPLC/MS,which ensure the quality of Jiangya Kangxian Formula in subsequent experiments.(3)The results of animal experiments showed that,compared with WKY group,the blood pressure of SHR group increased significantly(P<0.01),the cardiac function index decreased(P<0.01),and the levels of serum myocardial enzymes increased(P<0.05,P<0.01).The level of inflammatory factors was increased(P<0.05,P<0.01).Compared with SHR group,KXF treatment could effectively reverse the above indicators(P<0.05,P<0.01).(4)11 active ingredients of Jiangya Kangxian Formula were identified through HPLC/MS.The result of molecular docking and functional enrichment analysis indicated that Jiangya Kangxian Formula might play a role in treating hypertensive myocardial fibrosis by regulating mitochondrial function and oxidative stress reaction.As evidence,Jiangya Kangxian Formula could repair the function of damaged mitochondria,and also inhibit oxidative stress reaction in SHR myocardium.(5)The result of gene chip revealed 137 differentially expressed gene in myocardial tissues after treatment with Jiangya Kangxian Formula.Functional enrichment analysis showed that Jiangya Kangxian Formula might improve myocardial fibrosis through regulating TGF-beta signaling pathway.According,Jiangya Kangxian Formula reduced the protein expressions in TGF-beta signaling pathway in SHR myocardium.Besides,Jiangya Kangxian Formula can inhibit the proliferation,migration and collagen secretion of cardiac fibroblasts,as well as the protein expressions in TGF-beta signaling pathway.Conclusion:Jiangya Kangxian Formula constructed by network pharmacology and TCM theory has therapeutic effect on hypertensive myocardial fibrosis in SHR.The mechanism may be related to the improvement of mitochondrial function,oxidative stress response and the inhibition of TGF-beta signaling pathway.