The Effect Of Qi Huang Decoction On Small Intestinal Mucosal Immunity And Mechanical Barrier After Gastrectomy In Rats

1 Objective In this exeriment,we established model rats of surgical excision of the stomach and intervened the rats with Qi Huang Decoction by dropping it in the intestinal cavity.We studied the effects of Qi Huang Decoction on small intestinal mucosal barrier.We revealed the truth that Qi Huang Decoction protected the function of intestinal mucosal barrier.The mechanism of QH on intestinal mucosal immune barrier and mechanical barrier function was preliminarily discussed.2 Methods 2.1 Molding and grouping Refer to our previous mature modeling methods.This study aims to observe the effect of Qihuang decoction on small intestinal mucosal barrier after gastrectomy in rats.Materials and methods: A total of 80 Wistar rats were randomly divided into normal group,sham operation group,enteral nutrition group(EN)and Qihuang decoction group(EN+QH),there were 20 rats in each group.2.2 Ingredient analysis and methodology This experiment uses UPLC technology to construct fingerprint atlas of different batches of astragalus decoction,obtain corresponding common peak data information,and lay a foundation for the research on quality control and material foundation of astragalus decoction.2.3 Intervention methods of each group Both the EN group and the EN+QH group underwent gastrectomy.Instillation of enteral nutrition in the small intestine was performed after operation in the EN group.Instillation of enteral nutrition and Qihuang decoction in the small intestine was performed after operation in the EN+QH group.Only the abdominal incision and closing was performed in the sham operation group without drug and nutritional intervention.2.4 The index of observation and its detection method Taking small intestine of rats after intervention in 1 week,the total length,collection of isolated small intestine lymphatic nodules(Peyer 's patches,Ps)cells,and lamina propria lymphocyte(lamina propria lymphocytes,LPs)and gut mucosal surface type secretory immunoglobulin A(secretory Ig A,s Ig A).2.4.1 Intestinal mucosal immune barrier index detection(1)The s Ig A content in different anatomic sites of intestinal mucosa was determined by double antibody-PEG radioimmunoassay technique.(2)The number of Ig A+B cells in different anatomic sites of intestinal mucosa was determined by immunohistochemical method.The expression levels of tight junction proteins in intestinal epithelial cells were determined by western blotting method.The number of Ig A+B cells in different anatomic sites of intestinal mucosa was determined by immunohisto chemical method.2.4.2 Detection of intestinal mucosal mechanical barrier index The expression levels of tight junction proteins(occludin?claudin-1?claudin-5?ZO-1?ZO-2?P-occludin?P-claudin-1?P-claudin-5?P-ZO-1?P-ZO-2)in intestinal epithelial cells were determined by western blotting method.3 Statistical method All analyses were conducted using SPSS 17.0 software(SPSS Inc.,Chicago,USA).The data were expressed as x±s.The homogeneity of variance was tested by Levene method in each group.If the variance is homogeneous,the single factor analysis of variance and t test are used to compare the sample means.The nonparametric test is used if the variance is not uniform.P values <0.05 were considered indicative of a significant difference.4 Result The results showed that there was no difference in the composition and efficacy between 10 drugs,and the main water-soluble components were baicalin.The s Ig A content in the sham operated group was significantly lower than that of normal group(P<0.05).The number of Ig A+B cells and s Ig A content in Peyer's patches and lamina propria lym-phocytes in the EN group were significantly lower than that of sham operation group(P<0.05,P<0.01,P<0.001),The number of Ig A+B cells in Peyer's patches and lamina propria lym-phocytes in the EN+QH group were significantly higher than that of EN group(P<0.01,P<0.05).Compared with normal group,the level of claudin-1was increased in sham operation group;The phosphorylation levels of occludin,claudin-1,claudin-5,ZO-1and ZO-2 increased in the EN group,while the expression levels of non-phosphorylated occluding,claudin-1,claudin-5,ZO-1 and ZO-2 proteins decreased in the EN group(P<0.01,P<0.05).After treatment of Qihuang decoction for 7 days,compared with EN group,the phosphorylation levels of occludin,claudin-1,claudin-5,ZO-1 and ZO-2 significantly decreased in the EN+QH group,while the expression levels of non-phosphorylated occluding,claudin-1,claudin-5,ZO-1 and ZO-2 proteins significantly increased in the EN+QH group(P<0.01,P<0.05).5 Conclusion Qihuang decoction can promote the proliferation and differentiation of Ig A+B lymphocytes and increase the s Ig A content in intestinal mucosal immune barrier after gastrectomy in rats,it also can promote the expression of tight junction proteins to improve the permeability of intestinal mucosa and promote the recovery of intestinal immune barrier dysfunction in rats after gastrectomy by inhibiting the tight junction associated proteins' phosphorylation.