论文部分内容阅读
AIM: To investigate the expression of toll-like receptor 4(TLR4) and MD-2 gene and protein in Kupffer cells (KCs)and their role in ischemia-reperfusion (IR) injury of rat liver graft.METHODS: KCs were isolated at 0 (control group), 2, 12,24 h (IR group) following IR in rat liver graft. mRNA expression of TLR4 and MD-2 was detected by RT-PCR analysis, protein expression of TLR4/MD-2 was detected by flow cytometric(FCM) analysis,and tumor necrosis factor-α (TNF-α) level in supatant was measured by ELISA.Then isolated KCs were incubated with anti-TLR4 polyclonal antibody (antiTLR4 group),and TNF- level was measured again. RESULTS:The mRNA and protein expression of TLR4/MD-2 and the level of TNF-α in IR group increased significantly at 2 h following IR,and reached the maximum at 12 h,and slightly decreased at 24 h,but were still significantly higher than those in the control group (P＜0.01＝.The expression of these factors was markedly decreased after anti-TLR4rnantibody treatment as compared with the IR group (P＜0.01＝.CONCLUSION:Lipopolysaccharide (LPS) following IR can up-regulate TLR4/MD-2 gene and protein expression in KCs,and synthesize cytokine TNF-α.Anti TLR4 antibody can inhibit the production of TNF-α induced by LPS.TLR4 and its partner molecule MD-2 may play an important role in Kupffer cell activation and IR injury.rn