The Therapeutic Effect And Mechanisms Of Ginsenoside Rgl On Con A-induced Liver Injury

ObjectiveThis study was designed to investigate the therapeutic effect of Rg1and the potentialmechanisms on Con A-induced liver injury in mice.Methods1. C57BL male healthy mice were randomly divided into four groups.Group A(NSgroup) was the vehicle control, in which mice were not administrated Con A or Rg1, butthey were given saline100μl equally.Group B(Rg1group) was the Rg1control, in whichmice were administrated Rg1(5mg/ml,100ul)pretreatment by tail vein, without Con Atreatment6hours after pretreatment. Group C (Con A group) was the Con A controlgroup, in which mice were injected Con A (4mg/ml,100μl)by tail vein, without Rg1pretreatment. Group D (Rg1/Con A group) was the treatment group, in which mice wereinjected Rg1(5mg/ml,100ul) by tail vein with6hours before being administrated ConA(15mg/kg,100μl).2. Blood was harvested18h after pretreatment. Plasma was detached to measure ALTand AST by automatic biochemical analyzer, which was to assess the degree of liver damage.3. Livers of mice were harvested18h after pretreatment.It was to assess the degreeof liver damage by liver pathology.4. Blood of mice was harvested18h after pretreatment. Plasma was detached tomeasure levels of TNF-α, IL-6and IFN-γ by ELISA.5. Livers of mice were harvested18h after pretreatment. It was to detect mRNAlevels of TNF-α、IFN-γ、IL-6、CXCL-10、ICAM-1、NF-κB in liver by RT-PCR.6. Livers of mice were harvested18h after pretreatment. It was to detect proteincontent of phospho-p65、phospho-IκBα in liver by Westenblot.7. Mice blood, livers and spleens were harvested18h after pretreatment to detectcell counts of CD4~+T lymphocytes, CD8~+T lymphocytes by Flow cytometry. Results1. The ALT, AST levels of mice in Con A group were significantly higher than theother three groups (p<0.01).But it didn’t show difference among NS group, Rg1group andRg1/Con A group (p>0.05).2. The treatment of mice with Rg1pretreatment before Con A can significantlyreduce the incidence of liver necrosis. The pathological scores showed that liver injury inCon A group was more serious than that in Rg1/Con A group (p<0.05).3. Compared with mice in Con A group,ELISA showed levels of TNF-α、IFN-γ、IL-6in Rg1/Con A group were lower than that in Con A group.(p<0.01).4. Compared with mice in Con A group,RT-PCR showed TNF-α、IFN-γ、IL-6、CXCL-10、ICAM-1、NF-κB mRNA expression of Rg1/Con A group were lower than thatin Con A group in liver.(p<0.01).5. Compared with mice in Con A group, Westernblot showed protein content ofphospho-p65and phospho-IκBα in Rg1/ConA group were lower than that in ConA groupin liver.6. Liver CD4~+T cell counts of Rg1/Con A group were lower than Con A group(p<0.01),but CD8~+T lymphocytes cell counts between Rg1/ConA group and ConA groupwere not much difference (p>0.05).CD8~+T lymphocytes and CD4~+T lymphocytes cellcounts of blood, spleens didn’t show difference amoung the four groups (p>0.05).Conclusion1. Ginsenosides Rg1alleviating the severity of hepatic inflammation via suppressingpro-inflammatory TNF-α, IFN-γ,IL-6secretion by downregulating expression of NF-κBin liver.2. Ginsenoside Rg1reducing CD4~+T lymphocyte infiltration in liver may bemediated via inhibition of the expression of adhesion ICAM-1and chemokine CXCL-10.