CAJ | 학술논문

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-/一和内皮细胞TLR4+/+、髓系细胞TLR4+/+和内皮细胞TLR4+/+、髓系细胞TLR4+/+和内皮细胞TLR4-/-、髓系细胞TLR4-/-和内皮细胞TLR4-/-4组嵌入体或纯合体小鼠,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.榆测血清淀粉酶含量,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TLR4 mRNA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±8.084 E-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.
목적 탐색수계세포(중성립세포、혈소판)급내피세포표면Toll양수체4(toll-like receptor 4,TLR4)표체재소서급성배사성이선염(ANP)중성립세포초모중적작용.방법 장C3H/He-J화C3H/He-N소서통과골수이식적방법제작수계세포TLR4-/일화내피세포TLR4+/+、수계세포TLR4+/+화내피세포TLR4+/+、수계세포TLR4+/+화내피세포TLR4-/-、수계세포TLR4-/-화내피세포TLR4-/-4조감입체혹순합체소서,령설1개대조조.복강주사와피소、미정맥주사지다당복제ANP모형.유측혈청정분매함량,행이선병리검사、이선조직내분AS-D록을산지매염색계수、수과양화물매(MPO)활성측정、TLR4단백표체검측급RT-PCR검측외주혈립세포TLR4 mRNA표체.결과 각실험조소서혈청정분매균교대조조현저승고,단조간무현저성차이.4개실험조이선병리분치분별위5.52±1.21、5.18±1.02、2.03±0.82、1.92±0.78;이선MPO수평분별위(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-D계수분별위(66.88±2.17)개、(75.00±2.43)개、(21.50±2.38)개、(20.00±2.19)개;외주혈립세포TLR4 mRNA표체량분별위0.037±1.047E-2、1.489±8.084 E-2、1.470±5.210E-2、0.017±6.668 E-3;내피세포TLR4+/+적2조소서이선내혈관내피세포TLR4단백강양성표체,내피세포TLR4-/-적2조불표체.내피세포TLR4+/+적2조소서적이선손상、MPO활성、AS-D계수급TLR4단백표체균현저고우내피세포TLR4-/-적2조소서.결론 내피세포이비외주혈립세포재ANP중성립세포취집급병리손상중기관건작용.
Objective To investigate the role of expression of tall-like receptor 4 (TLR4) on medullary system cell (neutrophii, platelet) and endothelial cell surface in polymorphonuclear neutrophils (PMN) recruitment in rata 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 lipopolysaccharide. 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. 210E-2, 0. 017 ± 6. 668 E-3, respectively; the 2 groups with endothelial eel1 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 the 2 groups with endothelial cell TLR4 +/+ were significantly higher than those in the 2 groups with endothelial cell TLR4-/-. Conclusions It was endothelial cell, not peripheral blood granulocyte, which played a key role in the process of neutrophil recruitment and pathological injure of ANP.