Integrated strategy of RNA-sequencing and network pharmacology for exploring the protective mechanism of Shen-Shi-Jiang-Zhuo formula in rat with non-alcoholic fatty liver disease

• Published in: Pharmaceutical biology 2022-12, Vol.60 (1), p.1819-1838
• Main Authors: Xu, Zheng, Wu, Fan-Wei, Niu, Xuan, Lu, Xiao-Peng, Li, Yan-Rong, Zhang, Shu-Ting, Ou, Jun-Zhao, Wang, Xue-Mei
• Format: Article
• Language: English
• Subjects: 1-Phosphatidylinositol 3-kinase; AKT protein; Alanine transaminase; Aspartate aminotransferase; CD36 antigen; Cholesterol; Collagen; Drug development; Fatty liver; Fibrosis; hepatic fibrosis; Hepatic lipid accumulation; Lipid metabolism; Liver diseases; liver function damage; Pharmacology; PI3K/AKT pathway; Ribonucleic acid; RNA; Therapeutic targets; Traditional Chinese medicine; Triglycerides; γ-Glutamyltransferase
• ISSN: 1388-0209; 1744-5116
• DOI: 10.1080/13880209.2022.2106250

Shen-Shi-Jiang-Zhuo formula (SSJZF) exhibits a definite curative effect in the clinical treatment of non-alcoholic fatty liver disease (NAFLD). To explore the therapeutic effect and mechanism of SSJZF on NAFLD. Sprague Dawley rats were randomly divided into control, NAFLD, positive drug (12 mg/kg/day), SSJZF high-dose (200 mg/kg/day), SSJZF middle-dose (100 mg/kg/day), and SSJZF low-dose (50 mg/kg/day) groups. After daily intragastric administration of NAFLD rats for 8 weeks, lipid metabolism and hepatic fibrosis were evaluated by biochemical indices and histopathology. Then we uncovered the main active compounds and mechanism of SSJZF against NAFLD by integrating RNA-sequencing and network pharmacology, and PI3K/AKT pathway activity was verified by western blot. High dose SSJZF had the best inhibitory effect on hepatic lipid accumulation and fibrosis in rats with NAFLD, which significantly down-regulated total triglycerides (58%), cholesterol (62%), aspartate aminotransferase (57%), alanine aminotransferase (41%) andγ-glutamyl transpeptidase (36%), as well as the expression of ACC (5.3-fold), FAS (12.1-fold), SREBP1C (2.3-fold), and CD36 (4.4-fold), and significantly reduced collagen deposition (67%). Then we identified 23 compounds of SSJZF that acted on 25 key therapeutic targets of NAFLD by integrating RNA-sequencing and network pharmacology. Finally, we also confirmed that high dose SSJZF increased p-PI3K/PI3K (1.6-fold) and p-AKT/AKT (1.6-fold) in NAFLD rats. We found for first time that SSJZF improved NAFLD in rats by activating the PI3K/Akt pathway. These findings provide scientific support for SSJZF in the clinical treatment of NAFLD and contribute to the development of new NAFLD drugs.