TLR4的表达与急性胆道梗阻时内毒素致小鼠肝脏损伤的关系  

LPS increases the liver injury via TLR4-dependent manner in mice with acute obstructive jaundice

作　　者：李敏[1,2] 殷莉波[1,2] 刘平果[2] 赵文秀[2] 王效民[2] 尹震宇[2] 高翔[3] 

机构地区：[1]厦门大学医学院,福建361005 [2]厦门大学附属中山医院 [3]南京大学动物模式研究所

出　　处：《肝胆外科杂志》2010年第3期224-227,共4页Journal of Hepatobiliary Surgery

摘　　要：目的探讨急性胆道梗阻时内毒素损伤小鼠肝脏的机制及其与TLR4表达的关系。方法雄性C57BL/10J(WT)小鼠42只,随机分为生理盐水组(NS组,n=21)、内毒素处理组1(LPS1组,n=21),C57BL/10ScnJ(TLR4-/-)小鼠21只,为内毒素处理组2(LPS2组)。3组均行胆总管结扎术,LPS1、LPS2组小鼠于胆总管内注射LPS(8ng/μL,10ng/g体重),NS组注射同等剂量的生理盐水,术后6、12、24h采集标本,RT-PCR检测肝脏组织TLR4mRNA的表达情况,全自动生化分析仪检测血清ALT、TBIL、DBIL水平,ELISA法检测血清TNF-α、IL-6的水平。病理观察肝脏损伤情况,免疫组织化学染色观察肝脏NF-κB的表达。结果 LPS1组与NS组比较肝脏组织TLR4mRNA在6h时表达已有升高,于24h达高峰,ALT、TBIL各时点均明显升高(P<0.01),TNF-α、IL-6表达亦增高(P<0.01),LPS1病理损伤程度较NS组重,免疫组化显示术后24小时NF-κB在LPS1组可见肝细胞明显的核表达。LPS2组与LPS1组比较各血清学指标均明显下降,病理损伤减轻,24h时肝细胞NF-κB核表达较少。结论在急性胆道梗阻时LPS可以加重肝脏组织损伤和机体炎症反应,可能与TLR4的表达增高及NF-κB的表达有关。阻断LPS-TLR4信号通路可以减轻LPS引起的机体损伤。Objective To investigate whether LPS increase the liver injury and involved mechanism in mice with obstructive jaundice.Methods 42 of C57BL/10J（WT）male mice were randomly divided into NS group（n=21）and LPS1 group（n=21）,and LPS2 group as C57BL/10ScnJ（TLR4-/-）male mice（n=21）.All mice were subjected to common bile duct ligation（CBDL）,then the LPS1 and LPS2 groups were injected with LPS（8 ng/μL,10 ng/g body weight）via common bile duct after CBDL operation;the NS group was injected with equal amount of saline.The blood samples and liver tissues were collected at 6,12,24 h（n=7）after treatment and prepared for further analysis.The concentrations of serum ALT 、TBIL、DBIL were measured by automatic biochemistry analyzer;the levels of serum TNF-α and IL-6 were measured by enzyme-linked immunosorbent assay;the expressions of TLR-4 mRNA in the liver tissue were detected by RT-PCR;histological analysis showed the injury of the liver tissue and the distribution of NF-κB were detected by immunohistochemical staining.Results Liver tissue injury was more severe in the LPS1 group compared with the NS and LPS2 groups.mRNA level of TLR-4 in liver tissues was increased 6 hours after CBDL and LPS injection in LPS1 group,and peaked at 24 hours;No detection of TLR4 mRNA in LPS2 group;TNF-α and IL-6 secretions were significantly increased （P〈0.01）in LPS1 group;NF-κB was obviously expressed in the nucleus of hepatic cells in the LPS1 group at 24 hours after surgery,and slightly expressed in the NS group and LPS2 group.Conclusion LPS increased the liver injury after mice were subjected to CBDL.The activation of NF-κB and release of TNF-α and IL-6 were correlated with the higher expression of TLR4 in the liver and might be involved in this process.

关 键 词：胆道梗阻 LPS TOLL-LIKE receptor4 

分 类 号：R657.43[医药卫生—外科学]