卡巴胆碱对脓毒症大鼠肠道上皮细胞通透性和紧密连接的作用

发布时间：2018-06-12 05:46

本文选题：卡巴胆碱 + 脓毒症　；参考：《广东医学》2016年16期

【摘要】：目的探讨卡巴胆碱对脓毒症大鼠肠道上皮细胞通透性和紧密连接的作用。方法取健康成年雄性SD大鼠,随机分为4组(每组8只),经腹腔注射10 mg/kg脂多糖(LPS)复制脓毒症模型:对照组:腹腔注射等体积的生理盐水;脓毒症组:腹腔注射10 mg/kg LPS;卡巴胆碱组:腹腔注射10 mg/kg LPS,15 min后腹腔注射0.1mg/kg卡巴胆碱;α-BGT组:腹腔注射10 mg/kg LPS,5 min后皮下注射1.0μg/kgα-BGT,10 min后腹腔注射0.1 mg/kg卡巴胆碱。给予LPS 3 h后放血处死动物,取回肠组织,光镜下观察肠道上皮组织损害,荧光分光光度计测量体外漏出的FITC-Dextran,统计肠系膜淋巴结、肝、脾中细菌移位数量,电镜下观察肠道上皮细胞紧密连接超微结构变化,共聚焦显微镜下观察肠道上皮细胞紧密连接ZO-1和claudin-2蛋白移位,Western blot法检测紧密连接ZO-1和claudin-2蛋白含量。结果给予LPS后3 h,各组大鼠肠黏膜上皮细胞严重损伤,微绒毛坏死、脱落,炎性细胞浸润伴出血,肠黏膜组织学评分显著升高(P0.05),肠道上皮组织通透性显著升高(P0.05),肠系膜淋巴结、肝、脾中的菌落数明显增加(P0.05),肠道上皮细胞紧密连接结构破坏,为低密度电子致密影,肠道上皮细胞紧密连接ZO-1和claudin-2蛋白结构缺失,细胞周围着色减少,大量移位。腹腔注射卡巴胆碱后可显著减轻脓毒症大鼠上述改变,皮下注射α-BGT后可逆转卡巴胆碱的上述作用。结论卡巴胆碱可减轻脓毒症大鼠肠道上皮细胞损伤,降低其通透性,改善上皮细胞紧密连接蛋白移位和表达,该作用是通过激活胆碱能抗炎通路完成的。
[Abstract]:Objective to investigate the effect of Kaba choline on the permeability and close connection of intestinal epithelial cells in rats with sepsis. Methods the healthy adult male SD rats were randomly divided into 4 groups (8 rats in each group), and 10 mg/kg lipopolysaccharides (LPS) were injected into the abdominal cavity to replicate the sepsis model: the control group was injected into the abdominal cavity with equal volume of saline, and the sepsis group was injected 10 mg/kg intraperitoneally. LPS: Kaba choline group: intraperitoneal injection of 10 mg/kg LPS, 15 min after intraperitoneal injection of 0.1mg/kg Kaba choline; alpha -BGT group: intraperitoneal injection of 10 mg/kg LPS, 5 min after subcutaneous injection of 1 micron g/kg alpha -BGT, 10 min after the abdominal injection of 0.1 Kaba choline. FITC-Dextran was measured in vitro by spectrophotometer. The number of bacterial translocations in mesenteric lymph nodes, liver and spleen were counted. Ultrastructural changes were observed under electron microscopy. Under confocal microscopy, the intestinal epithelial cells were closely linked to ZO-1 and Claudin-2 egg white shift, and ZO-1 and Claudi were closely linked by Western blot method. The content of n-2 protein was 3 h after LPS. The intestinal mucosal epithelial cells were seriously damaged, microvilli necrosis, shedding, inflammatory cell infiltration and bleeding, intestinal mucosa histology score increased significantly (P0.05), intestinal epithelial tissue permeability increased significantly (P0.05), intestinal mesangial lymph node, liver and spleen increased significantly (P0.05), intestinal epithelium The cell closely connected structure was destroyed by the low density electron dense shadow, the intestinal epithelial cells closely connected ZO-1 and Claudin-2 protein structure loss, the peripheral coloring of the cells decreased, and a large amount of displacement. After intraperitoneal injection of Kaba choline, the above changes in septic rats could be significantly reduced, and the above effect of Kaba choline could be reversed after subcutaneous injection of alpha -BGT. Conclusion card Choline alleviates intestinal epithelial cell damage in septic rats, reduces its permeability and improves the displacement and expression of close connexin in epithelial cells, which is accomplished by activating the cholinergic anti-inflammatory pathway.
【作者单位】：郑州大学第五附属医院消化内科;河南省人民医院消化科;
【分类号】：R459.7

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