Screening-Based Optimization of a Herbal Mixture (JH01) with Robust Anti-Obesity Effects in High-Fat Diet-Induced Obesity

, using a screening-based analytical approach combined with experimental validation. Individual herbal extracts and their mixture were screened at 100 and 500 μg/mL in 3T3-L1 adipocytes. Based on superior anti-adipogenic efficacy, JH01 was selected for further study. Its effects were evaluated in vitro by Oil Red O staining and quantitative real-time PCR analysis of adipogenic genes, and in vivo using a high-fat diet (HFD)-induced obese mouse model, assessing body weight, serum lipid profiles, liver function markers, adipokine levels, and hepatic histology. JH01 showed markedly stronger inhibition of lipid accumulation than individual herbal components. JH01 significantly suppressed adipocyte differentiation and downregulated PPARγ, C/EBPα, and SREBP-1 expression in 3T3-L1 cells. Furthermore, JH01 modulated inflammatory cytokines and adipokine levels, as evidenced by reduced TNF-α, IL-6, and IL-1β levels and increased adiponectin levels. In HFD-fed mice, JH01 reduced body weight gain, serum triglyceride and total cholesterol levels, improved ALT and AST levels, decreased leptin concentrations, and attenuated hepatic steatosis. JH01 exerts potent anti-obesity and hepatoprotective effects through coordinated regulation of lipid metabolism and adipogenesis, supporting its potential as a multi-herbal therapeutic strategy for obesity-related metabolic disorders.