Semaglutide (HY-114118, MCE) is suitable for in vivo experiments. A recommended reference (PMID: 35143711) investigated the inhibitory effect of Semaglutide on non-alcoholic steatohepatitis (NASH) in a GAN diet-induced obese (DIO) mouse model of NASH. The experimental details are as follows:

• Treatment group: Semaglutide (30 nmol/kg/day, subcutaneous injection, once daily),.
• Control group: Vehicle (physiological saline with 0.1% bovine serum albumin, subcutaneous injection, once daily).
• Normal control group: Mice fed a normal chow diet, subcutaneous injection of physiological saline.
• Treatment duration: 8 and 12 weeks to assess short-term and long-term effects, respectively.
• Dose escalation protocol: To minimize initial adverse effects caused by GLP-1 receptor agonists (such as taste aversion), Semaglutide dosage was gradually increased from 0.6 nmol/kg following the sequence 0.6 → 1.2 → 2.4 → 4.8 → 12 → 30 nmol/kg, reaching the target dose on day 6 and maintained until the end of the study.

• Blood biochemistry: Measurement of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides (TG), total cholesterol (TC), among others.
• Liver histology: Liver tissues were fixed and paraffin-embedded, followed by hematoxylin and eosin (H&E) staining to evaluate steatosis, inflammation, and ballooning degeneration, and Picrosirius Red (PSR) staining to assess fibrosis. Automated scoring of NAS and fibrosis staging were performed using a deep learning tool (GHOST).
• Molecular level analysis: RNA sequencing of liver tissues was conducted to analyze transcriptomic changes related to inflammation, fibrosis, and metabolism-associated gene expression.

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