Effect And Its Potential Mechanism Of Prophylactic Satidroside In Mice With Concanavaline A-induced Liver Injury

ObjectiveThis study was aimed to investigate the effect of prophylactic salidroside (SAL) andits potential mechanisms on liver injury in mice induced by Con A via measuring thecytokines, pathological changes, protein content, etc.Methods1. C57BL/6male healthy mice were randomized divided into three groups. Normalsaline (NS) group, in which mice were administrated with normal saline100μl first and1h later with another dose of100μl normal saline. Concanavaline A (Con A) group, inwhich mice were administrated with normal saline100μl, and1h later with Con A(20mg/kg,100μl). Salidroside (SAL) group, in which mice were administrated withsalidroside (50mg/kg,100ul) via tail vein, and1h later were administrated with ConA(20mg/kg,100μl).2. Mice (n=30) in the three groups were anesthetized with sevoflurane12h after thelast injection. Blood was harvested via heart puncture and the plasma was collected tomeasure the level of alanine aminotransferase (ALT) and aspartate aminotransferase (AST)by automatic biochemical analyzer. Livers of mice were captured and stained with thehematoxylin and eosin (HE) reagent according to standard protocols. The severity ofliver injury was observed under light microscopy.3. Mice (n=30) in the three groups were anesthetized with sevoflurane12h after thelast injection. Blood of mice was collected using2ml work drum via heart puncture andthe plasma was used to measure the level of TNF-α, IFN-γ, IL-6and IL-10byenzyme-linked immunosorbent assay (ELISA); Livers of mice were captured and grinded.The mixture fuils of grinde livers was used to detect mRNA levels of TNF-α, IFN-γ, IL-6,IL-10and CXCL-10via RT-PCR.4. Mice (n=30) in the three groups were anesthetized with sevoflurane12h after thelast injection. Blood was harvested via heart puncture. Livers and spleens of mice wereharvested and grinded. All samples of mice blood, liver and spleens were used tomeasure cell counts of CD4+, CD8+T lymphocytes by flow cytometry.5. Mice (n=30) in the three groups were anesthetized with sevoflurane12h after the last injection. Livers of mice were captured and used to measure NF-κB p65and IκBαprotein content via westen-blot.Results1. The levels of ALT and AST in Con A groups were dramatically higher than that inNS groups (p<0.01). The levels of ALT and AST were decreased significantly in SALgroups (p<0.01). Nevertheless, no difference was detected between NS group and SALgroup (p>0.05). Pathologicial changes including massive and submassive tissue necrosis,dedma, and cell necrosis were detected via HE staining under light microscopy in theCon A group and the pathological scores in Con A group was significantly higher thanthat in NS group. However, the pathological changes were attenuated by salidroside viaH&E staining under light microscopy. The pathological scores in SAL group wassignificantly lower than that in Con A group (p<0.01) and higher in NS group (p<0.01).2.The number of CD4+T and CD8+T lymphocytes measured by flow cytometrywas significantly lower in Con A group compared with NS group in blood and spleen(p<0.01) and higher in liver (p<0.01). The number of CD4+T and CD8+T lymphocytesdidn’t show any difference in SAL group compared with Con A group in blood (p>0.05),but was significantly lower in liver (p<0.01) and higher in spleen (p<0.01). The numberof CD4+T and CD8+T lymphocytes was significantly lower in SAL group comparedwith NS group in blood and spleen (p<0.05), and the number of CD4+T lymphocyteswas significantly higher (p<0.01) while CD8+T lymphocytes didn’t show anydifference (p>0.05).3. The levels of TNF-α, IFN-γ, IL-6and IL-10measured by ELISA weresignificantly higher in Con A group compared with NS group (p<0.01) and significantlylower in SAL group compared with Con A group (p<0.01). The levels of TNF-α, IFN-γ,IL-6and CXCL-10mRNA measured by RT-PCR were significantly higher in Con Agroup compared with NS group (p<0.01) and IL-10mRNA was dramatically lower inCon A group compared with NS group (p<0.01). The expression of mRNA of TNF-α,IFN-γ, IL-6and CXCL-10was significantly lower in SAL group compared with Con Agroup (p<0.01), while IL-10mRNA was dramatically higher in SAL group comparedwith Con A group (p<0.05).4. Expression of NF-κB p-65and IκBα protein were decreased in SAL group compared wirh Con A group demonstrated by western-blot.ConclusionSalidroside could ameliorate Con A induced liver injury by reducing CD4+T andCD8+T lymphocyte infiltration in liver. Salidroside could downregulate the expressuionof CXCL-10and inhibit inflammatory of TNF-α, IFN-γ, IL-6and IL-10secretion, whichmight via attenuating the expression of NF-κB p-65and IκBα protein...