【摘要】 目的 探讨肠型脂肪酸结合蛋白对脓毒症大鼠早期急性肠损伤的诊断意义。方法 将120只大鼠随机分为假手术组、3h脓毒症模型组、6h脓毒症模型组、12h脓毒症模型组,每组各30只。采用盲肠结扎穿孔术(CLP)制备脓毒症大鼠模型,假手术组大鼠仅开腹而不行CLP。各组大鼠于造模后经尾静脉推注生理盐水1ml/100g液体复苏。将其分别装入大鼠代谢笼内收集尿液。3h、6h、12h后取血并处死大鼠,采用酶联免疫吸附试验(ELISA)检测血清内毒素、肠型脂肪酸结合蛋白(IFABP)、D-二乳酸(D-LAC)及尿IFABP表达;采用细菌培养方法培养淋巴结、肝脏、脾脏细菌移位量;光镜下观察肠黏膜组织病理学改变进行chiu评分;采用原位末端缺刻记法(TUNEL)检测肠黏膜上皮细胞凋亡指数(AI)。采用蛋白质免疫印迹法(Western Blot)检测IFABP蛋白、ZO-1蛋白表达。结果 与假手术组比较,M3、M6、M12组中血、尿IFABP、内毒素和FITC-G表达水平明显增高,差异有统计学意义(P<0.01),且M3、M6、M12各组间相互比较有统计学意义(P<0.01);而M3 D-LAC表达水平增高不明显,与假手术组无统计学意义,M6、M12组D-LAC表达水平明显增高,与假手术组有统计学意义(P<0.01),且各组间有统计学意义(P<0.01);细菌移位表达:3h淋巴结内发现有细菌移位,6h肝脏发现细菌移位,而脾脏在本实验时间窗内未发现细菌移位。6h淋巴结内细菌移位比3h明显(P<0.01),12h淋巴结、及肝脏内细菌移位率进一步增多,与6h比较有统计学意义(均P<0.01)。HE染色和肠黏膜细胞凋亡:3h、6h、32h肠黏膜损伤评分及细胞凋亡明显增高,与空白对照组比较有统计学意义(均P<0.01),6h肠黏膜损伤评分及细胞凋亡比3h更显著(均P<0.01),12h肠黏膜损伤评分及细胞凋亡进一步增多,与6h相比有统计学意义(均P<0.01);ZO-1、IFABP蛋白表达:M3、M6、M12中ZO-1表达水平逐渐减少,而IFABP蛋白表达水平逐渐增高,与假手术组有统计学意义(均P<0.01),且各组间比较有统计学意义(均P<0.01)。结论 IFABP对脓毒症大鼠急性肠损伤有较好的相关性,其对急性肠损伤的诊断有较好的敏感性和特异性。更多还原

【Abstract】 Objective To discuss intestinal fatty acid binding protein for the diagnosis in septic rats with acute intestinal injury.Methods Severe sepsis was established by cecal ligation arid puncture(CLP) in 30 healthy Sprague-Dawley rats. Thirty rats that received sham surgery received 10 ml/kg of normal saline. Rats(n=90) with CLP were randomized to receive 10 ml/kg of normal saline. Rats were killed after 3 hours, 6 hours and 12 hours.Serum levels of endotoxin, intestinal fatty acid binding protein, D-lac and urine intestinal fatty acid binding protein were measured by enzyme-linked immunosorbent assay. The lymph node, liver, spleen were bacteria cultured by bacteria culture assay. Ileum tissue structure and pathological score were observed by microscopy. Ileal mucosal epithelial cell apoptosis index was calculated by TUNEL assay. And tight junction transmembrane protein zo-1 and intestinal fatty binding protein were measured by immunohistochemistry and immunoblot. Results The level of serum and urine intestinal fatty acid binding protein, endotoxin, bacteria culture of the lymph node, FITC-G, ileum pathological score, apoptosis index of ileal mucosal epithelial cells, and intestinal fatty acid binding protein level were significantly higher in the M3 group than in the sham group(P<0.01).The level of serum and urine intestinal fatty acid binding protein, endotoxin, D-LAC, bacteria culture of the lymph node and liver FITC-G, ileum pathological score, apoptosis index of ileal mucosal epithelial cells, and intestinal fatty acid binding protein level were significantly higher in the M6 group than in the M3 group(P<0.01). The level of serum and urine intestinal fatty acid binding protein, endotoxin, D-LAC, bacteria culture of the lymph node and liver, FITC-G, ileum pathological score. apoptosis index of ileal mucosal epithelial cells, and intestinal fatty acid binding protein level were significantly highest in the M6 group(P<0.01), ZO-1 protein lever were significantly lower in the M3 group than the sham group(P<0.01).ZO-1 protein lever were significantly lower in the M6 group than the M3 group(P<0.01). ZO-1 protein lever were significantly least in the M12 group.Conclusion IFABP has a very good correlation for acute intestinal injury with sepsis rat, its good value to diagnosis acute intestinal injure.更多还原