Application Of Dicliptera Chinensis (L.) Juss (Acanthaceae) Polysaccharide In The Preparation Of Drugs For Immune Liver Injury

Objective: In this study,Lipopolysaccharide(LPS)was used to establish an injury model in vivo and in vitro.The aims of this study are to investigate the protective effect of Dicliptera chinensis(L.)Juss(Acanthaceae)polysaccharide(DCP)on LPS-induced immune liver injury,and to explore the effects of DCP on the NF-κB inflammatory pathway and the Fas/Fas L ligand system,in order to find a new method to improve immune liver injury.Methods: Sixty mice were randomly divided into six groups: control group,model group,silymarin group(150mg/kg)and DCP dose group(200,100,50 mg/kg).The immune liver injury model was established by intraperitoneal injection of LPS.Hematoxylin and Eosin(H&E)staining was used to observe the pathological changes of liver tissues.Reagent kits of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total bilirubin(TBIL),malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were employed for biochemical detection of serum.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)in the liver were detected by enzyme-linked immunosorbent assay(ELISA).The expression levels of p-NF-κBp65 and p-IKKα/β in liver were detected by immunohistochemistry.RT-PCR was used to detect the m RNA expressions of NF-κB and Fas/Fas L pathways.Western blot was used to detect protein expression levels of p-NF-κB,TNF-α,IL-6,IL-1β,Fas,Fas L,FADD,Caspase-8,Caspase-3,Bax and Bcl2.The LO2 cells were divided into normal group,LPS group and DCP dose group(0.125,0.25,0.5 mg/m L).Immunofluorescence was used to detect the expression level of NF-κBp65 in LO2 cells interfered with LPS.RT-PCR was used to detect the m RNA expression levels of NF-κB and Fas/Fas L pathway.Western blot was used to detect protein expression levels of p-NF-κB,TNF-α,IL-6,IL-1β,Fas,Fas L,FADD,Caspase-8,Caspase-3,Bax and Bcl2.Results:(1)DCP has a protective effect on LPS-induced immune liver injury in mice,and can significantly reduce liver cell necrosis.(2)ELISA results showed that DCP significantly reduce the expression of inflammatory cytokines TNF-α,IL-6 and IL-1β,and inhibit the inflammatory response to protect the liver.(3)The result of immunohistochemical showed that DCP can reduce the expression levels of p-NF-κBp65 and p-IKKα/β in the liver.(4)The immunofluorescence result showed that DCP could reduce the expression of NF-κBp65 in LO2 cells interfered with LPS.(5)DCP can inhibit NF-κB and Fas/Fas L pathways and significantly down-regulated NF-κB,TNF-α,IL-6,IL-1β,Fas,Fas L,FADD,Caspase-8,caspase-3,and Bax,and up-regulate the expression of Bcl2.Conclusions: DCP can alleviate LPS-induced immune liver injury by reducing the release of inflammatory cytokines in the liver,inhibiting inflammation response and cell apoptosis.