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

Abstract

Context Shen-Shi-Jiang-Zhuo formula (SSJZF) exhibits a definite curative effect in the clinical treatment of non-alcoholic fatty liver disease (NAFLD). Objective To explore the therapeutic effect and mechanism of SSJZF on NAFLD. Materials and methods 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. Results High dose SSJZF had the best inhibitory effect on hepatic lipid accumulation and fibrosis in rats with NAFLD, which significantly down-regulated t otal 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. Discussion and conclusion 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.

Widespread exposure to SARS-CoV-2 in wildlife communities

“…We performed a detailed computational investigation into the impact of the E 471 V and G 798 D mutations for the structure of the S protein using molecular modelling and molecular mechanics/generalized borne surface area (MM/GBSA) free energy calculations. The open conformation structure of S protein trimer from the glycosylated BA.2 Omicron variant was used for residue variant mapping [PDB 7XO8; (14)]. Because the crystal structure shows the trimer bound to the human ACE2 (hACE2) receptor, it is possible to investigate the impact of mutations in the hACE2 RBD and predict favorable interactions between SARS-CoV-2 S protein and hACE2 as previously reported (15) (Fig.…”

“…Our focus was on examining the local impact of E 471 V and G 798 D on the favorability of S protein-hACE2 interaction and the modifications in the S2 site as a hypothesis for the experimentally observed traits found in SARS-CoV-2 wildlife reservoir. Here, the open conformation structure of the S protein trimer from the glycosylated BA.2 Omicron variant was used for residue variant mapping [PDB 7XO8] 23 . Because the crystal structure shows the trimer bound to the human ACE2 (hACE2) receptor, it is possible to investigate the local impact of mutations in the RBD on interactions between SARS-CoV-2 S protein and the human ACE2, as a prediction for mechanistic impact on S protein-ACE2 interaction favorability as previously reported and validated experimentally 24 (Fig.…”

“…Computational studies were carried out using the structure of the glycosylated Omicron BA.2 variant bound to the human ACE2 receptor [Protein Data Bank 7XO8; (16)]. We incorporated point mutations in BA.2 using PyMOL (The PyMOL Molecular Graphics System v. 2.4.0, Schrödinger, LLC) to generate a new structure containing both E471V and G798D mutations.…”

Chinese medicine in the treatment of non-alcoholic fatty liver disease based on network pharmacology: a review

“… Wu, et al (2023b) found through network pharmacological study that The liver protective effect of QTHX is closely related to oxidative stress, anti-inflammatory response, and lipid receptor signaling, and QTHX has been shown to treat liver inflammation in rats by activating the SCOS1/NF-κB/TLR4 pathway. Xu, et al (2022) conducted a comprehensive analysis of Shen-Shi-Jiang-Zhuo formula (SSJZF) by RNA sequencing and network pharmacology, and found 23 main compounds of SSJZF and 25 key therapeutic targets for NAFLD. SSJZF was proven to improve NAFLD in rats by activating the PI3K/Akt pathway.…”