内毒素耐受导致的GSK-3抑制对急性内毒素性肝损伤的保护机制.docVIP

PAGE PAGE 8 内毒素耐受导致的GSK-3抑制对急性内毒素性肝损伤的保护机制 徐发良1，陈小乐1，朱宁生1，熊忠讯1，于 峰1，曾俊杰1，黄 春2 (400030 重庆，重庆市肿瘤研究所，重庆市肿瘤医院乳腺中心1；400010 重庆，重庆医科大学附属第二医院肝胆外科2) [摘要] 目的 探讨内毒素（lipopolysaccharide, LPS）耐受导致的糖原合成酶激酶-3(glycogen synthase kinase-3,GSK-3)活性抑制对急性内毒素性肝损伤产生保护效应的潜在机制。方法 建立LPS和GSK-3抑制剂氯化锂（LiCl）预处理的SD大鼠模型，采用半定量RT-PCR检测肝组织肿瘤坏死因子-alpha（TNF-alpha）和白细胞介素-10（IL-10）mRNA表达水平，以专用试剂盒检测代表肝组织中性粒细胞浸润程度的髓过氧化物酶（MPO）活性；用密度梯度离心法分离Kupffer细胞，建立LPS耐受和GSK-3抑制剂预(LiCl和胰岛素)处理的Kupffer细胞模型，用中性红法测定细胞吞噬活性；采用激光共聚焦显微镜和Western blot实验观察Kupffer细胞核因子-kapa B(NF-κB) p65的亚细胞定位和核转位情况。结果 LPS耐受和GSK-3抑制剂LiCl预处理均可减少大剂量LPS攻击导致的肝组织TNF-alpha mRNA上调表达（P0.05）、促进IL-10 mRNA上调表达（P0.05），减少LPS攻击导致的MPO活性升高（P0.05）；LPS耐受和GSK-3抑制剂（LiCl和胰岛素）均能增强Kupffer细胞吞噬活性（P0.05）；LPS耐受和GSK-3抑制剂LiCl预处理均可减少大剂量LPS导致的NF-kapaB p65核转位（P0.05）。结论 LPS耐受导致的GSK-3功能活性抑制，可能通过对NF-kapaB p65核转位、炎症细胞因子分泌、中性粒细胞浸润和Kupffer细胞吞噬活性等下游事件产生影响，从而保护急性内毒素性肝脏损伤。 [关键词] 糖原合成酶激酶-3 (GSK-3);内毒素或脂多糖(LPS)；肝脏；氯化锂；炎症性损伤；器官保护. [中图法分类号] R392.32; R34 [文献标志码] A The protective mechanism of LPS tolerance related GSK-3 inhibition on endotoxin induced acute liver injury in rats with endotoxemia XU Faliang, CHEN Xiaole, ZHU Ningsheng， XIONG Zhongxun, YU Feng, ZENG Junjie， HUANG Chun (1. Center of Breast, Chongqing Cancer Hospital and Institute, Chongqing 400030 China; 2. Department of Hepatobiliary Surgery, Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010 China) [Abstract] Objective To probe into the protective mechanism of LPS tolerance related GSK-3 inhibition on endotoxin induced acute liver injury in rats with endotoxemia. Methods The SD rat model of LPS tolerance or GSK-3 inhibitor LiCl pretreatment was established, and mRNA levels of TNF-alpha and IL-10 in hepatic tissue were analysed by semi-quantitative RT-PCR. To evaluate the degree of hepatic neutrophils infiltration, the activity of myeloperoxidase (MPO) in liver tissue was detected by using special kit. Kupffer cells were isolated by Ficoll density gradient centrifugation, and the ce