芪黄煎剂对胃切除术后大鼠肠黏膜上皮细胞间紧密连接的影响

作者	单位
李子义，于庆生，彭辉，沈毅，张琦，余树山，张万宗	1.安徽中医药大学第一附属医院普外一科（合肥 230031）；2. 安徽省中医药科学院中医外科研究所（合肥 230031）；3. 安徽中医药大学第一附属医院西区普外科（合肥 230031）

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中文摘要:

目的探讨芪黄煎剂对胃切除术后大鼠肠黏膜上皮机械屏障紧密连接（TJs）形态、结构和其相关蛋白表达的影响。方法 SD大鼠90只随机分为6组，空白对照组不做任何处理；假手术组仅行腹部剖开及缝合处理，术后不做任何处理；模型组胃切除术后予肠内营养；芪黄煎剂低、中、高剂量组在胃切除术后除肠内营养外，给予低、中、高剂量 [5、 10、 20 g/（ kg · d）]的芪黄煎剂小肠内滴注；每组15只，干预1周。透射电子显微镜下观察大鼠肠黏膜上皮TJs的形态、结构及宽度；激光共聚焦显微镜下观察TJs相关Claudin-1、Occludin和闭锁小带蛋白（ZO-1）的分布和荧光强度；蛋白质印迹检测Claudin-1、Occludin和ZO-1表达水平。结果空白对照组TJs表现出较强的连续性和完整性；假手术组、模型组TJs宽度较空白对照组变窄（P<0.01），模型组TJs宽度较假手术组变窄（P<0.01）；芪黄煎剂3组TJs宽度较模型组增宽（P<0.01），且呈剂量依赖性。与空白对照组比较，假手术组和模型组Claudin-1蛋白荧光强度和表达增加（P<0.05，P<0.01），Occludin和ZO-1蛋白荧光强度和表达降低（P<0.05，P<0.01）。与模型组比较，芪黄煎剂3组Claudin-1、Occludin和ZO-1蛋白荧光强度和表达显著增加（P<0.01），且呈剂量依赖性。结论芪黄煎剂能够促进胃切除大鼠肠黏膜上皮TJs形态和结构的修复，调节TJs蛋白Claudin-1、Occludin和ZO-1的表达。

英文摘要:

ABSTRACT Objective To investigate the effect of Qihuang Decoction（QHD） on the morphology，structure and related protein expression of tight junctions （TJs） in the mechanical barrier of intestinal mucosal epithelium in rats after gastrectomy. Methods Totally 90 SD rats were randomly divided into 6 groups，no intervention was conducted in blank control group； merely abdominal incision and suture with no postoperative intervention were conducted in sham-operated group；enteral nutrition was supplemented in model group after gastrectomy；QHD low，medium and high dose group were instilled QHD low，medium and high dose（5，10， 20 g·kg-1 · d-1）into small intestine in the base of enteral nutrition after gastrectomy；15 rats in each group，the intervention duration was 1 week. The morphology，width and integrity of TJs were observed by transmission electron microscope（TEM）. The distribution and fluorescence intensity of TJs related proteins Claudin-1，Occludin and zonula occludens 1（ZO-1） were detected by immunofluorescence technique. The expression of TJs related proteins Claudin-1，Occludin and ZO-1 were detected by Western Blot. Results The TJs in blank control group showed strong continuity and integrity，while the TJs width in the sham-operated group and model group was narrower than that in blank control group（P<0.01）. The TJs width in model group was further narrowed than sham-operated group（P<0.01），while the TJs width in the QHD groups was wider than model group and showed a dose-dependent manner（P<0.01）. Compared with the blank control group，the fluorescence intensity and expression of Claudin-1 protein increased（P<0.05，P<0.01），and the fluorescence intensity and expression of Occludin and ZO-1 proteins decreased in sham-operated group and model group（P<0.05，P<0.01）. Compared with the model group，the fluorescence intensity and expression of Claudin-1，Occludin and ZO-1 proteins in the QHD groups increased and showed a dose-dependent manner（P<0.01） . Conclusion QHD can promote the recovery of shape and structure of intestinal mucosal epithelial TJs in gastrectomy rats，regulate the expression of TJs proteins Claudin-1，Occludin and ZO-1.

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