健脾通里中药芪黄煎剂对胃切除大鼠肠黏膜生物屏障损伤的影响

Effect of the Spleen-strengthening and Interior-penetrating Traditional Chinese Medicine Qihuang Decoction on Intestinal Biological Barrier Injury in Rats after Gastric Resection

目的观察健脾通里中药芪黄煎剂对大鼠胃切除手术创伤后肠黏膜生物屏障损伤的保护作用及其机制。方法将90只SD大鼠随机分成空白对照组,假手术组,肠内营养组,芪黄煎剂低、中、高剂量组,每组15只。采用胃切除术复制大鼠肠黏膜损伤模型。空白对照组和假手术组术后自由饮水进食,不给予肠内营养液和中药;肠内营养组术后注射肠内营养液;芪黄煎剂低、中、高剂量组分别按照每日5、10、15 g/kg剂量滴注中药。光学显微镜下观察各组大鼠肠上皮绒毛形态和结构,采用凝胶法检测大鼠体内内毒素含量,运用细菌培养技术对大鼠结肠内容物乳酸杆菌、双歧杆菌、大肠杆菌、肠球菌培养与计数。RT-PCR荧光定量检测紧密连接蛋白(zonula occluding-1,ZO-1)、封闭蛋白(claudin)、咬合蛋白(occludin)的mRNA表达水平。结果芪黄煎剂能显著降低肠黏膜损伤大鼠门静脉、肠道和肠系膜淋巴结中内毒素水平(P<0.05),显著增加肠道双歧杆菌、乳酸菌、肠杆菌、肠球菌数量(P<0.05),显著降低肠黏膜损伤Chiu氏病理评分(P<0.05),显著上调小肠组织中ZO-1、claudin、occludin及其mRNA表达水平(P<0.05),且其效应呈现明显的剂量依赖性。结论大鼠胃切除手术创伤可以引起肠道黏膜生物屏障的损伤,高剂量芪黄煎剂可以有效减轻肠黏膜损伤,更好地调节肠道菌群,提高肠黏膜机械屏障肠上皮细胞的ZO-1、claudin、occludin的蛋白表达水平,起到保护肠道黏膜屏障的作用。

Objective To investigate the protective effect of the spleen-strengthening and interior-penetrating traditional Chinese medicine Qihuang Decoction against intestinal biological barrier injury in rats after surgical trauma caused by gastric resection and its mechanism.Methods A total of 90 Sprague-Dawley rats were randomly divided into blank control group,sham-operation group,enteral nutrition group,and low-,middle-,and high-dose Qihuang Decoction groups,with 15 rats in each group.A rat model of intestinal mucosal injury was established by gastric resection.The rats in the blank control group and the sham-operation group were allowed to eat and drink freely after surgery,without enteral nutrient solution ortraditional Chinese medicine;the rats in the enteral nutrition group were given injection of enteral nutrient solution after surgery;the rats in the low-,middle-,and high-dose Qihuang Decoction groups were given dripping of the traditional Chinese medicine at a dose of 5,10,and 15 g/kg,respectively,every day.The morphology and structure of intestinal epithelial villi were observed under an optical microscope;the gel method was used to measure the content of endotoxin in rats;the bacterial culture technique was used for the culture and counting of Lactobacillus,Bifidobacterium,Escherichia coli,and Enterococcus in colonic contents;RT-PCR was used to measure the mRNA expression levels of zonula occluding-1(ZO-1),claudin,and occludin.Results Qihuang Decoction significantly reduced the level of endotoxin in the portal vein,intestinal tract,and mesenteric lymph nodes of rats with intestinal mucosal injury(P<0.05),significantly increased the number of intestinal Bifidobacterium,Lactobacillus,Enterobacter,and Enterococcus(P<0.05),significantly reduced the Chiu's pathological score of intestinal mucosal injury(P<0.05),and significantly upregulated the protein and mRNA expression levels of ZO-1,claudin,and occludin in small intestinal tissue(P<0.05),in a significant dose-dependent manner.Conclusion The surgical trauma of gastric resection may cause intestinal biological barrier injury in rats,and high-dose Qihuang Decoction can effectively alleviate intestinal mucosal injury,better regulate the change in intestinal flora,and increase the protein expression levels of ZO-1,claudin,and occludin in the intestinal epithelial cells of intestinal mucosal mechanical barrier,thereby exerting a protective effect on intestinal mucosal barrier.