慢性胰腺炎患者胰腺和血清中TLR4信号通路关键因子的检测及其意义  

Detection of key factors of TLR4 signaling pathway in pancreas and serum of patients with chronic pancreatitis and its significance

作　　者：田珍珍[1] 朱杰[2] 王淑华[3] 李雅君[1] 刘永方[1] 修明[1] 高润平[1] 

机构地区：[1]吉林大学第一医院肝胆胰内科,吉林长春130021 [2]山东大学齐鲁医院感染症科,山东济南250001 [3]吉林大学第一医院胃肠外科,吉林长春130021

出　　处：《吉林大学学报（医学版）》2013年第4期763-767,I0004,共6页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81070370;81270544)

摘　　要：目的:检测慢性胰腺炎(CP)患者胰腺和血清中Toll样受体4(TLR4)信号通路关键应答基因的表达,阐明TLR4信号通路在CP发生发展过程中的意义。方法:收集24例CP和6例胰腺正常的良性病变患者的手术胰腺组织标本,分别进行TLR4mRNA原位杂交和髓样分化蛋白88(MyD-88)、肿瘤坏死因子α(TNF-ɑ)免疫组织化学染色,利用电脑分析系统测量各个指标的积分光密度(IA)值。采集84例CP患者及30例正常成人的空腹静脉血,ELISA法检测血清可溶性TLR4(sTLR4)、TNF-α、白细胞介素6(IL-6)、白细胞介素12(IL-12)的水平,分光光度法检测血浆内毒素脂多糖(LPS)的水平。分析血浆LPS水平与各细胞因子的相关性。结果:与正常对照组比较,CP组患者胰腺组织TLR4mRNA、MyD-88、TNF-ɑ表达水平均显著增加(P<0.01),分别是正常对照的4.2、6.1及10.2倍;末梢血LPS、sTLR4、TNF-ɑ、IL-6和IL-12水平与正常对照组比较均升高(P<0.05或P<0.01);Bivariate相关分析,伴有内毒素血症的CP患者血浆LPS水平与血清TNF-α、IL-6、IL-12水平呈正相关关系(r=0.859,P<0.01;r=0.688,P<0.01;r=0.539,P<0.05)。结论:LPS-TLR4信号通路及应答基因产物在CP患者胰腺及外周血中均上调,其在CP急性发作期和炎症进展过程中发挥重要作用。Objective To detect the expression of key factors of toll-like receptor 4（TLR4）signaling pathway in pancreas and serum of the patients with chronic pancreatitis（CP）,and to explore the significance of TLR4signaling pathway in the occurrence and development of CP.Methods Pancreas tissues from 24patients with CP and 6controls with normal pancreatic histology were collected to detect TLR4 mRNA by in situ hybridization.The MyD88and TNF-ɑwere detected by immunohistochemistry.Computer image analysis was performed to measure integrated optimal density（IA）of TLR4mRNA,MyD88and TNF-ɑin pancreas sections.The fasting vein blood samples were collected in 84patients with CP and 30normal individuals.The serum soluble TLR4（sTLR4）,TNF-ɑ,IL-6and IL-12levels were measured by ELISA.The lipopolysaccharide（LPS）level in plasma was determined by spectrophotometry.The relationships between the plasma PLS level and different cytokines were analyzed.Results In situ hybridization analysis showed that the expression levels of TLR4mRNA,MyD88and TNF-ɑin the pancreas tissue of the CP patients were significantly higher than those in normal control group（P 0.01）,and they were 4.2-fold,6.1-fold and 10.2-fold of the normal controls.The blood LPS,sTLR4,TNF-ɑ,IL-6,and IL-12levels in the CP patients were significantly increased compared with normal control group（P0.05 or P0.01）.Bivariate correlation analysis showed that there were significantly positive correlations between the level of plasma LPS and the levels of serum TNF-α,IL-6and IL-12in the patients with endotoxemic CP（r=0.859,P0.01;r=0.688,P0.01;r=0.539,P0.05）.Conclusion LPS-TLR4signaling pathway and their response gene products in pancreas tissue and peripheral blood of the CP patients are up-regulated,and it plays an important role in acute attack stage and inflammation progression of CP.

关 键 词：慢性胰腺炎 脂多糖 内毒素血症 TLR4信号通路 应答基因 

分 类 号：R576[医药卫生—消化系统]