The Protective Effects Of Ilicifolius Alkaloid A And Its Derivates In Rats With Immunological Liver Fibrosis Induced By Porcine Serum

Objective: To investigate the anti-fibrosis effects and potentialmechanisms of Acanthus ilicifolius alkaloid A and its two derivates (TC-2, TC-3)in rats with liver fibrosis induced by porcine serum (PS).Methods:60Sprague Dawley (SD) rats were randomly divided into2groups: normal control group (n=10) and immunological liver fibrosis modelgroup (n=50). Normal control group received physiological saline and liverfibrosis model group received0.5ml PS via intraperitoneal injection twiceweekly for12weeks. After8weeks, the successful liver fibrosis models wereestablished and then randomly divided into5groups: model control group,colchicines (Col) group (0.4mg·kg-1), HBOA group (100mg·kg-1), TC-2group(100mg·kg-1) and TC-3group (100mg·kg-1). Four drugs were administrated byintragastric gavage once daily respectively. After4weeks treatment, blood wasdrawn from inferior caval vein. The levels of hyaluronic acid (HA) and laminin(LN) were measured by enzyme linked immunosorbent assay (ELISA) method. Liver tissues were excised and weighed for calculating liver index. The liverswere stained with HE to observe the liver tissues pathology. And then the degreeof liver damage was examined. The PCI, PCIII, TIMP-1, MMP-13, TLR4,MyD88, NF-κB mRNA expression of each group in rat liver tissues weredetected by RT-PCR. The TLR4, NF-κBp65, COX-2protein expression of livertissues were detected by immunohistochemistry.Results: Serological experimental results showed that, compared with thenormal control group, the levels of HA and LN were increased significantly inmodel group (P<0.01); compared with the model group, the levels of HA andLN in HBOA, TC-2and TC-3groups were decreased (P<0.05or P<0.01).Pathological results showed that HBOA, TC-2and TC-3groups could alleviatethe liver injury induced by PS. RT-PCR experiments showed that the expressionof PCI, PCIII, TIMP-1, TLR4, MyD88and NF-κB mRNA in model group ratswere increased significantly compared with normal control group (P<0.05orP<0.01), the expression of MMP-13mRNA was decreased significantly(P<0.05); compared with model group, the expression of PCI, PCIII, TIMP-1,TLR4, MyD88and NF-κB mRNA in HBOA, TC-2and TC-3groups weredecreased obviously (P<0.05or P<0.01), the expression of MMP-13mRNAwas increased significantly (P<0.05). Immunohistochemical experimentalresults showed that the protein expression of TLR4, NF-κBp65, COX-2inmodel group were increased compared with normal control group (P<0.05);compared with model group, the protein expression of TLR4, NF-κBp65,COX-2in HBOA, TC-2and TC-3groups were decreased significantly (P<0.05).Conclusion: Our data suggest that HBOA and its derivates can efficientlyinhibit PS-induced hepatic fibrosis in rats and the protective effect maybe due to the inhibition of ECM and regulation of TLR4、NF-κB and expression of COX-2.These may account for their hepatoprotective effect on hepatic fibrosis inducedby PS. However, TC-3has better anti-fibrosis effect in the three compounds.