## 3 Results  
### Confirmation of FRVE Detoxification and Main Components (Fustin and Fisetin) of FRVE  
To confirm FRVE detoxification, the presence of the allergen urushiol was determined by high-performance liquid chromatography (HPLC). Urushiol was not detected, thereby confirming the successful detoxification of FRVE (Figure 1), whereas the urushiol peak appeared in the standard at 21.818 min. Figure 1: HPLC chromatograms of FRVE. Peaks of two main components (fustin (1) and fisetin (2)) and allergen urushiol. Mobile phase: 0.1% (_v/v_) Formic acid in water (solvent A) and 100% (_v/v_) Methanol (solvent B) at a flow rate of 0.7 mL/min, with gradient as follows: 0–17 min, 100% B; 17–20 min, 100% B; 20–23 min, 0% B; 23–30 min, 0% B. and detected at 254 nm. Fustin (PubChem CID 5317435) and fisetin (PubChem CID 5281614) were the main active components present in FRVE (Figure 1). The peaks showed retention times of fustin and fisetin as 14.501 min and 16.778 min, respectively. FRVE Decreased Body Weight, Intra-Abdominal Fat Weight, and Liver Weight in the HFD-Induced NAFLD Model  
The bodyweights of the NCD, HFD-fed, FRVE, and MT (milk thistle extract)-treated groups were measured once a week for 13 weeks. As shown in Figure 2A, the bodyweight in the HFD-fed control group increased significantly after two weeks of feeding when compared to the bodyweight of the NCD group. In contrast, the FRVE and MT treated groups abrogated the bodyweight gain from the third week of FRVE and MT treatment. Eleven weeks after treatment, the mean bodyweights of HFD-fed control group (\(49.1\pm 1.18\) g) were notably higher than that of the NCD group (\(29.75\pm 1.03\) g). However, treatment with FRVE and MT suppressed the bodyweight to \(44.6\pm 2.07\) g and \(44.76\pm 2.65\) g, respectively (Figure 2A). The mean intra-abdominal fat pad weights were significantly high in HFD-fed mice (\(1.8\pm 0.31\) g) as compared to the NCD group mice (\(0.25\pm 0.02\) g). In contrast, FRVE and MT-treated mice exhibited decreased intra-abdominal fat pad weights of \(0.97\pm 0.09\) g and \(1.15\pm 0.34\) g, respectively (Figure 2B). Furthermore, HFD-fed mice presented an increased liver size and solid pink-colored livers. The liver weights of HFD-fed mice (\(2.58\pm 0.3\) g) were 1.53 times higher than the NCD group. In contrast, despite being on HFD, the FRVE (\(1.69\pm 0.07\) g) and MT-treated (\(1.63\pm 0.03\) g) groups showed decreased liver weights and size of bright pink color, similar to the NCD group (Figure 2C) (Supplementary Figure S1). Figure 2: Effect of FRVE on the body, intraabdominal fat, and liver weights in HFD-induced NAFLD model. (**A**) Bodyweight. * \(p<0.05\), ** \(p<0.01\) and *** \(p<0.001\) (compared to HFD control group). (**B**) intraabdominal fat. (**C**) liver weight. Data are expressed as mean \(\pm\) SD. * \(p<0.05\) (compared to HFD control group). *** \(p<0.001\) (compared to NCD control group). FRVE Downregulated Serum Liver Function and Lipid Markers in an HFD-Induced Nonalcoholic Fatty Liver Model  
As shown in Figure 3A,B, activities of the hepatotoxicity markers alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were significantly increased in the HFD-fed mice group. FRVE treatment downregulated the enhanced ALT and AST activities induced by HFD. The levels of ALT and AST were lower in the FRVE treated group than in the positive control group. In addition, FRVE treatment inhibited the HFD-induced elevation in serum total cholesterol (TC) and triglycerides (TG) levels (Figure 3C,D). FRVE Inhibited Hepatic Steatosis and Lipid Accumulation in an HFD-Induced Nonalcoholic Fatty Liver Model  
We confirmed HFD-induced hepatic steatosis in liver tissue sections by H&E staining. Histological analysis revealed a significant increase in the typical histological features of a nonalcoholic fatty liver, such as macro- or micro-vesicular fatty changes, and large swollen hepatocytes with ballooning degeneration in HFD-fed mice (Figure 4A).