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Abstract

Study on the Mechanism of Zhenwu Decoction in Decreasing the Fats in Non-Alcoholic Fatty Liver Disease Using System Pharmacology and Cytological Validation Test

Author(s): C. J. Zhang, Z. Y. Fang, Yu Qin Tang, L. Zheng, Hai Yan Zhu, Lili Huang, Lu Yun Xia, Wen Ying Huai, Q. Z. Yin, Yan Qiu Wang and Tian E Zhang*
Department of Basic Medical Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, Wenjiang 611137, 1Key Biology Laboratory for Traditional Chinese Medicine Zang-Xiang Research of Sichuan University, Chinese Medical Center of Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 610075, China

Correspondence Address:
Tian E Zhang, Key Biology Laboratory for Traditional Chinese Medicine Zang-Xiang Research of Sichuan University, Chinese Medical Center of Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 610075, China, E-mail: zhte2003@cdutcm.edu.cn


Purpose of this study was to screen the active compounds of Zhenwu decoction and their gene targets in the treatment of non-alcoholic fatty liver disease by network pharmacology and cytological validation test. The candidate compounds and related targets of Zhenwu decoction were obtained from traditional Chinese medicine system pharmacology database and PharmMapper. The non-alcoholic fatty liver disease-related genes were obtained from online Mendelian inheritance in man, GeneCards and DisGeNET databases. Molecular docking was carried out to simulate the binding affinities between potential core compounds and key target genes, and the chemical constituents of Zhenwu decoction were analyzed and identified by Ultra-Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry technology. Therapeutic effect and mechanism of Zhenwu decoction on non-alcoholic fatty liver disease were validated using in vitro analysis. The protein-protein interaction network analysis identified the albumin, protein kinase B1, epidermal growth factor receptor, caspase 3 and peroxisome proliferator activated receptor gamma as the key target genes. Gene ontology annotation analysis showed that the Zhenwu decoctionnon- alcoholic fatty liver disease target genes were mainly involved in the response to steroid hormone and lipid catabolic process. The results of the Kyoto encyclopedia of genes and genomes pathway enrichment analysis were mainly related to the Forkhead box O signaling pathway and phosphatidylinositol 3-kinaseprotein kinase B signaling pathway. Paeoniflorgenone had the highest binding affinities for albumin, protein kinase B1, epidermal growth factor receptor, caspase 3 and peroxisome proliferator activated receptor gamma. The in vitro experiment showed that 19.5 mg/ml (p≤0.05) and 39 mg/ml (p≤0.01) of Zhenwu decoction could reduce lipid accumulation. Real-time quantitative polymerase chain reaction analyses revealed that Zhenwu decoction induced down-regulation of epidermal growth factor receptor messenger ribonucleic acid levels and up-regulation of heat shock protein 90 alpha family class a member 1, mitogen-activated protein kinase 1 and phosphatidylinositol 3-kinase messenger ribonucleic acid levels.

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