The Effects Of Acanthus Ilicifolius Alkaloid A And Its Derivatives On Adiponetin, PPARγ Expressions In Rats With Hepatic Fibrosis

Objective:This study was designed to evaluate the effects of Ilicifolius Alkaloid A (4-hydroxy-2-benzoxazolone, HBOA) and its acetylated derivatives 4-acetoxy-2-benzoxazolone (TC-2) and 3-acetyl-4-acetoxy-2-benzoxazolonc (TC-3), on adiponetin, PPARy expressions in rats with hepatic fibrosis.Method:Experimental used SD rats were randomly divided into two groups as normal control group, and CC14 induced chemical liver fibrosis model group. Liver fibrosis model group received 50% CC14 oil solution, with the volume of 1 ml kg-1 by intragastric administration, two times per week for the duration of 8 weeks to induce liver fibrosis. In addition, the normal control group received the same volume of physiological saline. After 8 weeks administration, they were randomly divided into model group, colchicine group, HBOA group, TC-2 group and TC-3 group. The dosage for colchicine was 100mg.kg-1 and 3 treatment groups (HBOA, TC-2, TC-3) received the same dosage as 100mgkg-1. Each group of rats was given related treatment once a day for 4 weeks. After 4 weeks treatment, the rats were sacrificed and liver tissues were accurately weighed to calculating liver index [rat liver weight (g)/rat body weight (g)] ×100. A portion of the liver was fixed for histopathology and the left lateral lobe of the liver was collected from each group for RT-PCR evaluation. We used immunohistochemical method to detect the protein expression of adiponetin, TGF-β1 and TLR-4, and to explore the protective effect of Acanthus ilicifolius alkaloid A and its derivatives on liver fibrosis and related mechanisms.Result:Histopathological results showed that HBOA and its acetylated derivatives can obviously alleviate the liver fibrosis degree induced by CC14. Moreover, compare with the model group, HBOA, TC-2, TC-3 dosage groups can obviously inhibit the mRNA expression of TNF-a and IL-6 (P<0.01).In addition, the mRNA expression of adiponetin and PPARy in treated groups was increased significantly compared with the model group (P＜0.01 or P<0.05). Immunohistochemical results showed that compared with the normal control group, TGF-β1 and TLR-4 protein expression was significantly greater in the model group (P＜0.01) and lower in the treated groups (P<0.01). In contrast, the protein expression of adiponetin was obviously increased in HBOA, TC-2, TC-3 group compared with the model group (P＜0.01).Conclusion:Our study indicates that compared with the model group, HBOA, TC-2 and TC-3 groups had significantly decreased the production of pro-inflammatory cytokines which in turn will decrease the proliferation of HSC. From the results, it can be suggested that HBOA and its derivatives have beneficial effects against liver fibrosis, and the mechanisms may be related to the inhibition of inflammatory cytokines adiponetin, TNF-a, IL-6, PPARy, TGF-β1 and TLR-4 and the regulation of their responses.