小鼠肝内源性损伤下TLRs在Kupffer细胞和肝窦内皮细胞中的表达

Expression of Toll-Like Receptor 2 in Liver Kupffer Cell and Sinsusoidal Endothelial Cell in Hepatic Endogenous Injury in Mice

目的 观察在小鼠内源性损伤模型下肝脏Kupffer细胞 (Kupffercell,KC)和肝窦内皮细胞 (sinusoidalendothelialcells,SEC)表面Toll样受体 2 (TLR2 )蛋白及mRNA表达。方法 在肝部分缺血 再灌注损伤模型下 ,采用原位灌注消化法分离并纯化KC和SEC ,用大鼠抗小鼠TLR2 IgG和异硫氰酸荧光素 (FITC)的二抗进行染色 ,流式细胞仪 (FCM)测定阳性细胞数 ,并用实时定量PCR(Real TimeRT PCR)检测两种细胞中TLR2 mRNA含量。结果 损伤组KCTLR2 的表达明显高于假手术组 ,蛋白质表达为 ( 9.19± 1.0 7) %vs ( 1.5 2± 0 .2 1) % ,P<0 .0 1;mRNA表达为 0 .5 4± 0 .77vs 2 .6 2± 2 .19,P<0 .0 5。SEC差异并无显著性意义。结论 在肝缺血 再灌注损伤中 ,小鼠肝KCTLR2

Objective To observe the synthesis of TLR 2 protein and its mRNA expression in Kupffer cells (KCs) and sinusoidal endothelial cells(SECs)Methods Thirty two BALB/c mice divided into two groups (operation group and false operation group) were used to prepare the model of partial hepatic ischemia/reperfusion (I/R) injury. After injury KCs and SECs were isolated with two steps situ perfusion technique. And these cells were dyed by rat anti mouse TLR 2 IgG and anti rat IgG 2b labeled with flurescein isothiocyanate (FITC). The sysnthesis of TLR 2 protein were determined by flow cytometric (FCM) analysis and real time reverse transcription polymerase chain reaction (Real Time RT PCR) analysis for gene expression. Results As for KCs: TLR 2 expression was significant higher in operation group, compared with false operation group 〔protein expression: (9.19±1.07)% vs (1.52± 0.21)%, P <0.01; gene expression: 0.54±0.77 vs 2.62±2.19, P <0.05〕. But there were no significant differences with expression in SECs. Conclusion Synthesis of TLR 2 protein and its gene expression increased in KCs in the mouse partial hepatic ischemia reperfusion injury.