Jia Guo,1,2,* Anhua Shi,1,3,4,* Yanhong Sun,1 Shunzhen Zhang,1 Xiaoyi Feng,1,3,4 Yifan Chen,1 Zheng Yao1,3,4 

1School of Basic Medical Sciences, Yunnan University of Chinese Medicine, Kunming, Yunnan, 650500, People’s Republic of China; 2Dongtai Hospital of Traditional Chinese Medicine, Dongtai, Jiangsu, 224200, People’s Republic of China; 3The Key Laboratory of Microcosmic Syndrome Differentiation, Education Department of Yunnan, Yunnan University of Chinese Medicine, Kunming, Yunnan, 650500, People’s Republic of China; 4Yunnan Key Laboratory of Integrated Traditional Chinese and Western Medicine for Chronic Disease in Prevention and Treatment, Yunnan University of Chinese Medicine, Kunming, Yunnan, 650500, People’s Republic of China

*These authors contributed equally to this work

Correspondence: Zheng Yao, School of Basic Medical Sciences, Yunnan University of Chinese Medicine, Kunming, Yunnan, 650500, People’s Republic of China, Tel +8618908719365, Email YaoYNUCM@163.com

Objective: In this study, we investigated the effect of Shenling Baizhu San(SLBZS), Quzhi Ruangan Fang(QZRGF) and Gexia Zhuyu Tang(GXZYT) on the intestinal flora of NAFLD rats through network pharmacology and experimental validation.
Materials and Methods: Protein-protein interaction, Gene Ontology (GO), and molecular docking were performed. Male Sprague-Dawley (SD) rats were divided into 6 groups: Normal, Model, SLBZS (7.2g/kg), QZRGF (27.72g/kg), GXZYT (28.8 g/kg) and positive control (Fenofibrate, 18mg/kg); the NAFLD model was established by High-fat diet. After one week of acclimatisation feeding consecutively, continuous gavage was given for 8 W and 12 W. Serum, liver and faeces were collected and biochemical and pathological indices were determined. The diversity and abundance of intestinal flora were also analyzed using 16S rDNA amplified sequencing.
Results: A total of 132 active ingredients were obtained from the screening results of SLBZS. A total of 202 active ingredients were obtained from the screening results of GXZYT. The screening results of QZRGF obtained 34 active ingredients. Nine common hub genes were screened from the PPI network. GO functional analysis reported that these targets were mainly closely related to the response to bacterial molecules. The molecular docking results indicated that the 11 core constituents in three compound prescriptions has good binding ability with MAPK1, AKT1, CASP3, FOS, TP53, STAT3, MAPK3.
Conclusion: The Chinese herbal compounds SLBZS, QZRGF and GXZYT may exert lipid-lowering effects through multi-components, multi-targets and multi-methods for the treatment of NAFLD while improving the diversity and abundance of the intestinal flora of the rats, and the best effect was achieved with SLBZS.

Keywords: network pharmacology, Shenling Baizhu San, Quzhi Ruangan Fang, Gexia Zhuyu Tang, NAFLD, intestinal flora