Mechanic study of Qushi Kaiyu decoction on non-alcoholic fatty liver disease model rats based on the inhibition of TLR4/NF-ĸB pathway

Background:The objective of this research was to examine the impact of the Chinese herbal formula Qushi Kaiyu(QSKY)on rats with non-alcoholic fatty liver disease(NAFLD)and its inhibitory effect on the TLR4/NF-ĸB pathway.Methods:NAFLD model rats was constructed through high-fat diet.Meanwhile,rats were treated with QSKY(6.4 g/kg)by gavage.The therapeutic effect of QSKY on NAFLD was assessed by testing body weight change,the liver index,lipid concentrations in blood,and antioxidant and inflammatory levels;assessing liver function;and performing pathological staining including hematoxylin-eosin and Oil Red O.The protein levels of key factors in the TLR4/NF-ĸB pathway(TLR4,MyD88,p65 and IKB)in rat liver tissue were determined using western blotting in order to explore the mechanism responsible for the therapeutic effects of QSKY in rats with NAFLD.Results:QSKY significantly reduced the liver index and body weight value;reduced triglyceride,cholesterol,alanine aminotransferase,and aspartate aminotransferase levels in NAFLD rats;improved the pathological changes,such as ballooning degeneration,fat accumulation,necrosis,and inflammation;elevated GSH-Px and superoxide dismutase activities and lowered malondialdehyde levels,indicating that QSKY enhanced the antioxidant capacity;and reduced inflammatory cytokine(IL-6,IL-1β,and TNF-α)levels.Western blotting results showed that QSKY significantly reduced TLR4,MyD88,and decreased the phosphorylation of IKB and p65 protein levels in the livers of rats with NAFLD.Conclusions:QSKY showed therapeutic effects on NAFLD and can alleviate oxidative stress and inflammation.This mechanism may be related to an improvement in TLR4/NF-ĸB pathway.

Qushi Huayu Decoction (祛湿化瘀方) Inhibits Protein and Gene Expression of Cathepsin B in HepG2 Cells Induced by Free Fatty Acids

Objective： To study the experimental efficacy of Qushi Huayu Decoction （祛湿化瘀方,QHD） on protein and gene expression of cathepsin B （ctsb） in HepG2 cells induced by free fatty acids （FFAs）.Methods： The model of HepG2 steatosis and tumor necrosis factor-α （TNF-α） secretion was induced by long-chain FFAs.HepG2 cells were divided into 4 groups： control group （group C）,model group （group M）,low-dose QHD group （group L） and high-dose QHD group （group H ）.Long-chain FFAs were added to groups M,L and H.The 10% blank-control serum was added to group C and M,while 5% and 10% QHD-containing sera were added to group L and H,respectively.The levels of serum TNF-α and cellular triglyceride （TG） were detected.Cellular p-IκB and ctsb expression were detected using Western blot and PCR.The expression and distribution of ctsb were observed by immunofluorescence.Results： After incubating with FFA for 24 h,TG deposition in HepG2,TNF-α content in cell supernatant,the protein expression of cellular ctsb and P-IκB,as well as mRNA expression of ctsb increased markedly in group M compared with group C （P〈0.05,P〈0.01）.Compared with group M,TG deposition,the expression of cellular ctsb,P-IκB and ctsb mRNA in groups L and H,as well as TNF-α content in group H,decreased significantly （P〈0.05）.Cell immunochemical fluorescence studies showed that ctsb was released from lysosomes and distributed in the cytoplasm extensively and diffusedly after being stimulated with FFA.In this study,these above-mentioned changes were inhibited markedly in groups L and H.Conclusion： QHD might have a direct inhibitory effect on the ctsb target in the FFA-ctsb-TNFα pathway of hepatic lipotoxicity.