HSP70 On The LPS Induced Acute Liver Injury

Subject:To investigate the effect of HSP70 on the LPS induced acute liver injury and LPS induced liver signal transduction.Methods:The experiment(Exp)1.1 SA 8mg/kg i.p.At 3,6,9,12,24h the liver was extirpated with carbrital anaesthesia and the proteins extracted from livers was assayed for the expression of HSP70.1.2 The Kunming mice were divided randomly into three groups.(1)control: 0.9%Sodium Chloride 0.2 ml,i.p..(2)LPS group:LPS 5mg/kg,i.p..(3)SA+LPS(SA)group:SA 8mg/kg,i.p.were administered on prenight 8pm then treated with i.p injection of LPS 5mg/kg,12h later.The blood was gathered from i.p.eye vein and liver was excised with carbrital anaesthesia after LPS or 0.9%Sodium Chloride injected at 0.Sh(n=4),1.5h(n=8)and 6.0h(n=10)for ALT, MDA,TNF-α,GSSG,GSH and GSH/GSSG assays for each groups.1.3.The animal and groups were same as Exp 1.2 in order to observe suvival rate o Exp 2.The experimental mice were divided randomly into two groups.(1)control:0.9%Sodium Chloride intraperitoneal injections.(2)SA. SA.intraperitoneal injections 8mg/kg.After 12h,India ink 1:5 diluted with germ free physiological saline was injected into the vail vein in each group rats.To calculate phagocytic index k and its correction a.Exp 3.The animal and groups were same as Exp 1.2(1)control:0.9%Sodium Chloride 0.2 ml,i.p..(2)LPS group:LPS 5mg/kg i.p..(3)SA+ LPS(SA)group:SA 8mg/kg,i.p.was administered on prenight(8pm),then treated with i.p injection of LPS 5mg/kg,12h later.The livers were excised in anaesthetic state by carbrital after LPS or 0.9%Sodium Chloride injected at 0.5h, 1.5h and 6h and the protein extracted from livers was assayed for the phosphorylation level of MEK1/2,ERK1/2,p38MAPK,STAT1,STAT3 and expression of TLR4,NF-κB,I-κB,CD14, IRAK-M and TNF-αby western blotting analysis.Results:HSP70 can expression and LPS-induced acute liver injury could attenuated significantly by SA pretreatment.The plasma ALT activity and liver MDA,GSSG contant were decreased remarkably by SA preteatment and the GSH and GSH/GSSG assays of liver was increased significantly.In LPS treatment groups,LPS stimulation increased plasma level of TNF-αexpression in the liver,in which the peak values of plasma TNF-αin SA group were much lower than LPS group(p＜0.05).HSP70 can increase survive rate of mice compare to LPS group(p＜0.05). CD14,TLR4,NF-κB,NF-αwere decreased significantly by SA pretreatment.And I-κB,p-MEK1/2,and p-ERK1/2,p-p38MAPK could increased.The phosphorylation of STAT1 was appeared to decreased after SA pretreatment,but the phosphorylation of STAT3 appear to no obviously change. Conclusion:SA pretreatment can induce expression of HSP70 of mice,partially inhibited LPS induced NF-κB signalling pathway and made TNF-αless releasing,to reduce LPS induced acute liver injury significantly.