Taohong Huazhuo decoction(桃 红化 浊汤)alleviates carbon tetrachloride-induced hepatic fibrosis in ratsby inhibiting the nicotinamide adenine dinucleotide phosphate oxidase 4/nucleotide-binding oligomerization domain-like...OA

OBJECTIVE:To explore the mechanism underlying the anti-fibrotic effects of Taohong Huazhuo decoction(THHZD,桃红化浊汤). METHODS:Forty percent carbon tetrachloride(CCl4)dissolved in olive oil(2 mL/kg)was intraperitoneally injected twice a week for 8 weeks to induce hepatic fibrosis in rats.The normal group received the same volume of olive oil alone.At week 5,rats injected with CCl4 were intragastrically administered normal saline,0.1 mg/kg colchicine,3.285 g/kg THHZD,6.57 g/kg THHZD,and 13.14 g/kg THHZD once a day for 4 weeks.A biochemical analyzer was used to detect the levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),and total bilirubin(TBIL).Hematoxylin and eosin(HE)staining and Masson staining were performed to evaluate pathological changes in the liver.Enzyme-linked immunosorbent assay(ELISA)was used to determine glutathione(GSH)and malondialdehyde(MDA)levels in the liver,and interleukin-1 β(IL-1β),and IL-18 in serum.The levels ofα-smooth muscle actin(α-SMA)and reactive oxygen species(ROS)in liver tissues were analyzed by immunohistochemistry and immunofluorescence,respectively.Western blot(WB)was used to analyze the expression levels of nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4),nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3),IL-1β,IL-18,caspase-1,and collagen Ⅰ in liver.Finally,in vitro experiments were conducted using hepatic stellate cells(HSC-T6). RESULTS:THHZD improved the pathological changes caused by CCl4 in the liver.Compared with the normal group,the levels of serum ALT,AST,TBIL,IL-1β,and IL-18 in the model group were significantly increased.NOX4,ROS,NLRP3,IL-1 β,IL-18,caspase-1,collagen Ⅰ,α-SMA,and MDA levels were increased in the liver,while GSH levels were decreased.The administration of THHZD to CCl4-exposed rats significantly reversed or eliminated the changes in the above-mentioned indicators.Moreover,THHZD inhibited transforming growth factor-β1(TGF-β1)-induced high expression of α-SMA and collagen Ⅰ in HSC-T6 cells,while downregulating the expression of proteins related to the NOX4/NLRP3 signaling pathway in vitro. CONCLUSION:THHZD reduced the activation of HSCs by inhibiting the NOX4/NLRP3 signaling pathway,thereby exerting anti-hepatic fibrosis effects.