Protective Effects Of Agmatine On Trauma-Hemorrhage Induced Acute Hepatic Injury

Objective: To observe protective effects of agmatine(AGM) on Trauma-Hemorrhage- induced liver injury in miceMethod:1. Forty-two adult male BALB/c mice were randomly divided into three groups( n = 14 each), including control group, model group( bilateral femoral fracture and removal of 35% of the total blood volume), and AGM group( trauma/hemorrhage +AGM 200 mg/kg). Seven mice in each group were sacrificed at 12 hours and 24 hours, respectively, after modeling,and blood samples and tissue homogenate of liver were collected and Isolation of Liver Kupffer Cells。2. Serum aspartate transaminase( AST), alanine aminotransferase( ALT) and lactic dehydrogenase( LDH) were determined with automatic biochemistry analyzer.Hepatic tissue was also collected for observation of pathological at 24 hours after modeling. The contents of tumor necrosis factor-α( TNF-α), interleukins( IL-6) in serum 、hepatic homogenate were determined with enzyme linked immunosorbent assay( ELISA).3. The contents of tumor necrosis factor-α( TNF-α), interleukins( IL-6) in kupffer supernatant were determined with enzyme linked immunosorbent assay( ELISA). Pro-inflammatory cytokine(TNF-α,IL-6) gene expression of the kupffer cell were assessed by RT-PCR。Western Blot detection kupffer cell cytoplasm expression of protein involved in the nf-kappa B, ELISA detection of kupffer cells nf-kappa Bp65 DNA binding forceResults:1.The control group normal liver tissue structure; At 24 hours after modeling, Histological examination demonstrated that AGM significantly reduced liver injury caused by trauma-hemorrhage, as manifested in amelioration 0f hepatocytes swelling,necrosis and neutrophil infiltration,at the same time AGM group can effective improvtion the pathological changes and reduction the serum ALT、AST、LDH.2.There was no significant difference in the levels of hepatic tissue TNF-αand IL-6 at 12 hours after modeling; Compared with control group,24 hours after trauma/hemorrhage,the levels of hepatic tissue and serum TNF-α and IL-6 in model group were significantly elevated,AGM can effective reduce the levels of hepatic tissue and serum TNF-α and IL-6[TNF-α( ng/mg:248.7±22.5vs.405.2±19.6, IL-6( ng/mg):22.5±3.1vs.58.4±7.7,all P<0.01);and [TNF-α(ng/l):58.2±3.1vs.80.8±4.7,IL-6(ng/l):74.1±6.6vs.104±9.0,all P<0.01)3.kupffer cell Model group in vitro TNF-α and IL- 6 levels higher than the control group obviously, AGM can effective reduce the levels of TNF-α and IL-6 in kupffer cell supernatant[TNF-α(ng/m L):50.4±4.8 vs.91.2±3.6,IL-6(ng/m L):386.2±24.6vs.609.5±23.5,all P<0.01],and also depressed the expression of m RNA TNF-α and IL-6. [TNF-α:4.3±0.3 vs.8.7±0.4,IL-6:3.2±0.3 vs.6.7±0.5,all P<0.01).Conclusion:1.AGM significantly reduced the level of ALT, AST and LDH in mice induced by trauma-hemorrhage and improved the liver pathological changes.AGM inhibited the prodution of inflammatory cytokine(TNF-αand IL- 6) in mice induced by trauma-hemorrhage2.AGM inhibited the prodution of inflammatory cytokine(TNF-αand IL- 6) of kupffer Cell induced by trauma-hemorrhage and Down-regulated the expression of m RNA(TNF-α and IL- 6)and Suppress the activation of kupffer cells NF- κB reduce the secretion of inflammatory cytokines.