Effect Of Acanthus Ilicifolius Alkaloid A On PI3k/Akt And NF-?B Signaling Pathways In Rats With Hepatic Fibrosis

Objective:This study was to investigate the effect of Acanthus ilicifolius alkaloid A?HBOA?on PI3K/Akt and NF-?B signaling pathways in rats with hepatic fibrosis induced by carbon tetrachloride?CCl₄?,exploring the underlying mechanism of HBOA against liver fibrosis.Methods:Rats were randomly divided into the normal group,model group,high,medium and low-dose HBOA-treated groups?100,50,25 mg/kg?,and colchicine group?0.4 mg/kg?.Except for the normal group,the rats in other groups were given 50%CCl₄ olive oil solution?2 mL/kg?twice a week for 12weeks to induce liver fibrosis.From the ninth week,the drug-administered group was given the corresponding test drug once a day for 4 weeks.At the end of treatment,the body change and liver index were calculated.The levels of alanine aminotransferase?ALT?and aspartate aminotransferase?AST?in the liver homogenate of each group were detected.The degree of hepatic fibrosis was observed by Sirius red staining.Serum IL-1?and IL-10 contents were detected by Enzyme linked immunosorbent assay?ELISA?.The protein expressions of NF-?Bp65,NF-?Bp50 and p-IKK?/?in liver tissue were determined by immunohistochemistry.The NF-?Bp65,NF-?Bp50,IL-6 and TNF-?mRNA expressions were measured by real-time fluorescence quantitative PCR.The protein expressions of NF-?Bp65,p-NF-?Bp65,NF-?Bp50,I?B?,p-I?B?,IKK?/?,p-IKK?/?,PI3K,p-PI3K,Akt,p-Akt,mTOR,p-mTOR,p70S6K,p-p70S6K,IL-6,TNF-?,ICAM-1 in liver tissue were detected by Western blot.Results:?1?HBOA improved liver function and and reduced collagen hyperplasiaThe experimental results showed that compared with the normal control group,the body weight of rats was significantly decreased,while the liver index,ALT and AST levels were increased in liver tissue in the model group?P<0.05?.Compared with the model group,the body weight of rats was significantly increased,and the liver index,ALT and AST levels were decreased in liver tissue in HBOA-treated groups?P<0.05?.Moreover,the results of Sirius red staining showed that the collagen fiber deposition was significantly reduced in HBOA-treated groups.These results demonstrated that HBOA improved liver function and and reduced collagen hyperplasia induced by CCl₄.?2?HBOA alleviated the inflammatory reactionELISA results showed that compared with the normal control group,serum IL-1?content was increased,while IL-10 content was decreased in the model group?P<0.05?.Compared with the model group,serum content of IL-1?level was decreased,serum content of IL-10 was increased in HBOA groups?P<0.05?.Moreover,Western blot and RT-PCR experimental results showed that compared with normal control group,the protein expression of IL-6,TNF-?and ICAM-1 as well as the expression of IL-6,TNF-?mRNA in liver tissue were significantly increased in model group?P<0.05?.Compared with the model group,the protein expression of IL-6,TNF-?and ICAM-1 as well as the expression of IL-6,TNF-?mRNA in liver tissue were significantly were significantly decreased in HBOA-treated groups?P<0.05?.These results demonstrated that HBOA alleviated the inflammatory reactionon in hepatic fibrosis rats.?3?HBOA inhibited the PI3K/Akt and NF-?B signaling pathwaysImmunohistochemistry and RT-PCR results showed that compared with normal control group,the protein expressions of NF-?Bp65,NF-?Bp50 and p-IKK?/?as well as the expressions of NF-?Bp65 and NF-?Bp50 mRNA in liver tissue were significantly increased in the model group?P<0.05?.Compared with the model group,the protein expression of NF-?Bp65,NF-?Bp50 and p-IKK?/?as well as the expressions of NF-?Bp65,NF-?Bp50mRNA in liver tissue were significantly decreased in HBOA-treated groups?P<0.05?.Moreover,Western blot experimental result showed that compared with normal control group,the phosphorylations of PI3K,Akt,mTOR,p70S6K,NF-?Bp65,I?B?and IKK?/?as well as the protein expression of NF-?Bp50in liver tissue were significantly increased in the model group?P<0.05?.Compared with the model group,the phosphorylations of PI3K,Akt,mTOR,p70S6K,NF-?Bp65,I?B?and IKK?/?as well as the protein expression of NF-?Bp50 were significantly decreased in the HBOA-treated groups?P<0.05?.These results demonstrated HBOA inhibited the PI3K/Akt and NF-?B signaling pathways.Conclusion:HBOA has a protective effect on hepatic fibrosis rats,and its mechanism may be associated with relieving inflammatory reaction by inhibiting the PI3K/Akt and NF-?B signaling pathways.