细胞表面Toll样受体4在小鼠急性坏死性胰腺炎中的作用  

Role of expression of toll-like receptor 4 （TLR4） on medullary system cell and endothelial cell surface in rats with acute necrotizing pancreatitis

作　　者：包俊炜[1] 吴河水[1] 张进祥[2] 徐颖[4] 张景辉[3] 田元[3] 王春友[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院胰腺外科,武汉430022 [2]华中科技大学同济医学院附属协和医院急诊外科,武汉430022 [3]华中科技大学同济医学院附属协和医院普外科实验室,武汉430022 [4]北京协和医院消化内科

出　　处：《中华胰腺病杂志》2009年第1期44-47,共4页Chinese Journal of Pancreatology

摘　　要：目的探索髓系细胞（中性粒细胞、血小板）及内皮细胞表面Toll样受体4（toll-like receptor 4，TLR4）表达在小鼠急性坏死性胰腺炎（ANP）中性粒细胞招募中的作用。方法将C3H／He—J和C3H／He．N小鼠通过骨髓移植的方法制作髓系细胞TLR4-／-和内皮细胞TLR4＋／＋、髓系细胞TIR4＋／＋和内皮细胞TLR4＋／＋、髓系细胞TLR4＋／＋和内皮细胞TLR4-／-、髓系细胞TLR4-／-和内皮细胞TLR4-／4组嵌人体或纯合体小鼠，另设1个对照组。腹腔注射蛙皮素、尾静脉注射脂多糖复制ANP模型。检测血清淀粉酶含量，行胰腺病理检查、胰腺组织萘酚AS-D氯乙酸脂酶染色计数、髓过氧化物酶（MPO）活性测定、TLR4蛋白表达检测及RT—PCR检测外周血粒细胞TLR4mRNA表达。结果各实验组小鼠血清淀粉酶均较对照组显著升高，但组间无显著性差异。4个实验组胰腺病理分值分别为5．52±1．21、5．18±1．02、2．03±0．82、1．92±0．78；胰腺MPO水平分别为（1．834±0．170）U／g、（2．596±0．138）U／g、（0．367±0．018）U／g、（0．202±0．018）U／g；AS—D计数分别为（66．88±2．17）个、（75．00±2．43）个、（21．50±2．38）个、（20．00±2．19）个．外周血粒细胞TLR4 mRNA表达量分别为0．037±1．047E-2、1．489±8．084E-2、1．470±5．210E-2、0．017±6．668 E-3；内皮细胞TLR4＋／＋的2组小鼠胰腺内血管内皮细胞TLR4蛋白强阳性表达，内皮细胞TLR4-／-的2组不表达。内皮细胞TLR4＋／＋的2组小鼠的胰腺损伤、MPO活性、AS—D计数及TLR4蛋白表达均显著高于内皮细胞TLR4-／-的2组小鼠。结论内皮细胞而非外周血粒细胞在ANP中性粒细胞聚集及病理损伤中起关键作用。Objective To investigate the role of expression of toll-like receptor 4 （TLR4） on medullary system cell （neutrophil, platelet ） and endothelial cell surface in polymorphonuclear neutrophils （PMN） recruitment in rats with acute necrotizing pancreatitis. Methods C3H/He-J mice and C3H/He-N mice were divided into 4 groups by bone marrow transplantation ： mosaic of medullary system cell TLR4-/- and endothelial cell TLR4 ＋ / ＋ ; mosaic of medullary system cell TLR4 ＋ / ＋ and endothelial cell TLR4-/-; mosaic of medullary system cell TLR4 ＋/＋ and endothelial cell TLR4 -/-; mosaic of medullary system cell TLR4-/- and endothelial cell TLR4-/-, another control group was also established. ANP was induced in all these groups by intraperitoneal injection of cerulein and caudal vein injection of lipopolysaceharide. Serum amylase, pancreatic tissue naphthol AS-D chloroacetate esterase, MPO activity, TRL4 mRNA expression in peripheral blood granuloeyte was determined by RT-PCR, TRL4 protein expression in pancreatic tissue was measured by immunohistochemisty. Results Compared with that of control group, the levels of serum amylase in the 4 groups all significantly elevated and there was no difference among these 4 groups. Pathological scores of pancreas in the 4 groups were 5.52 ± 1.21, 5.18 ± 1.02, 2.03 ± 0.82, 1.92 ± 0.78, respectively; MPO activities were （1.834 ± 0. 170） U/g, （2. 596 ± 0. 138） U/g, （0. 367 ± 0. 018） U/g, （0. 202 ± 0. 018） U/G, respectively ; AS-D counts were 66.88 ± 2.17, 75.00 ± 2.43,21.50 ± 2.38, 20.00 ± 2.19, respectively; the expressions of TLR4mRNA of granular cell in peripheral blood were 0. 037 ± 1. 047 E-2, 1. 489 ± 8. 084 E-2, 1.470 ± 5. 210 E-2, 0. 017 ± 6. 668 E-3, respectively; the 2 groups with endothelial cell TLR4 ＋/＋ had strongly positive expression of TLR4 protein in vascular endothelial cell; while the 2 groups with endothelial cell TLR4-/- had no expression; the pancreatic injuries, MPO activities, AS-D counts and TLR4 protein in

关 键 词：胰腺炎 急性坏死性 内毒素类 嵌合体 TOLL样受体 

分 类 号：R576[医药卫生—消化系统]