【摘要】 目的探讨脓毒症大鼠血清肠型脂肪酸结合蛋白(I-FABP)水平和在肠组织中的表达水平与急性肠损伤程度、内毒素血症及细菌移位的关系。方法将80只大鼠随机分为假手术组(n=40)和模型组(n=40),模型组采用改良的Zantl方法建立脓毒症大鼠模型,假手术组仅暴露升结肠。分别在造模后4、8和16 h各组取8只大鼠,采用EKT-5Mset动态革兰阴性菌试剂盒检测血清内毒素水平,采用ELISA试剂盒测定血清I-FABP及异硫氰酸荧光素标记蛋白-G(FITC-G)水平,通过细菌培养评估肝、脾、肾组织细菌移位情况,采用Western blotting和免疫组化SP法分析小肠组织中的I-FABP和磷酸化的血管扩张刺激磷蛋白(p-VASP)表达情况。采用苏木精伊红(HE)染色观察肠组织病理学变化。结果与假手术组相比,模型组大鼠血清I-FABP、FITC-G及内毒素水平均明显升高(P均<0. 05),且随时间的延长显著增加(P均<0. 05)。造模8 h后出现明显急性肠黏膜损伤,且随术后时间的延长而加剧。与假手术组比较,模型组大鼠的脾脏、肝脏和肾脏均出现明显细菌移位(P均<0. 05),并且各脏器内细菌移位的数量随时间的延长显著增加(P均<0. 05)。假手术组大鼠在手术后4、8和16 h时小肠组织中I-FABP和pVASP蛋白表达未见明显变化。模型组大鼠在手术后4、8和16 h小肠组织中I-FABP和p-VASP的蛋白表达较假手术组显著上调(P均<0. 05),并且表达水平随着时间的延长显著增加(P均<0. 05)。在手术后16 h时假手术组大鼠小肠组织中I-FABP和p-VASP蛋白轻度阳性表达,而模型组该两指标均呈强阳性表达。结论 I-FABP可作为脓毒症大鼠急性肠损伤程度和内毒素血症以及细菌移位的有效标志物。更多还原

【Abstract】 Objective To investigate the associations of serum intestinal fatty acid-binding protein(I-FABP) level and its expression level in intestinal tissues with the severity of acute intestinal injury,endotoxemia and bacterial translocation in septic rats. Methods A total of 80 rats were randomly divided into sham operation group(n = 40) and model group(n =40). Sepsis rat model were established by modified Zantl method in model group,and ascending colon was exposured only in sham operation group. At 4-,8-and 16-hour after modeling,8 rats were respectively taken from two groups to be detect.EKT-5 Mset dynamic gram-negative bacteria test kit was used to detect serum endotoxin level. Enzyme-linked immunosorbent assay(ELISA) kit was used to detect the levels of serum I-FABP and fluorescein isothiocyanate-labled protein-G(FITC-G). The situations of bacterial translocation in liver,spleen and kidney tissues were assessed by bacterial culture. Western blotting and immunohistochemistry SP method were used to detect the expressions of I-FABP and phosphorylated vasodilator-stimulated phosphoprotein(p-VASP) in intestinal tissues. HE staining was used to observe pathological changes of intestinal tissues. Results Compared with sham operation group,the levels of serum I-FABP,FITC-G and endotoxin increased significantly(all P < 0. 05),and they increased with the prolongation of time in model group(all P < 0. 05). Acute intestinal mucosal injury occurred 8 hours after modeling and was exacerbated with the prolongation of time. Compared with sham operation group,the bacterial translocation in liver,spleen and kidney was obvious(all P < 0. 05),and the quantity increased significantly with the prolongation of time(all P < 0. 05). The expression levels of I-FABP and p-VASP proteins in intestinal tissues at 4-,8-and 16-hour after operation remained unchanged in sham operation group(all P > 0. 05),while they were up-regulated significantly in model group compared with sham operation group(all P < 0. 05). At 16-hour after operation,the mild positive expressions of I-FABP and p-VASP proteins were seen in sham operation group,while their strong positive expressions were seen in model group. Conclusion I-FABP can be served as an effective maker of acute intestinal injury,endotoxemia and bacterial translocation in septic rats.更多还原