葛根芩连汤对肠道湿热证泄泻小鼠肠道机械屏障的影响OACSTPCD

Objective To explore the effects of Gegen Qinlian Decoction on the intestinal mechanical barrier in diarrhea mice with intestinal dampness-heat pattern.Methods Fifteen SPF male Kunming mice were randomized into normal group(n=5)and model group(n=10).The diarrhea model with intestinal dampness-heat pattern was replicated by the method of"high sugar and high fat+high temperature and high humidity+liquor dilution+ice water",and 10 successfully modeled mice were randomized into self-healing group and treatment group,with five mice in each group.The treatment group was administered with 5.07 g/(kg·d)Gegen Qinlian Decoction,and the normal group and self-healing group were given equal volumes of sterile water.The duodenum,jejunum,and ileum segments of mice in each group were aseptically extracted after 4 d of gavage.The general condition of the mice was observed;HE staining was used to observe the morphology of each intestinal segment.Image J processing software was used to measure the villus height(VH)and crypt depth(CD)in each group,and the ratio of VH to CD(V/C),the number of goblet cells,and the average optical density(AOD)value of tight junction protein were calculated.Two-step immunohistochemistry was used to analyze the expressions of Claudin 1,Occludin,and zonula occluden-1(ZO-1).Results The normal group of mice had shiny hair,good mental state,normal water intake and rectal temperature,and normal stool and urine.The treatment group of mice had white hair,good mental state,full stool particles,normally recovered rectal temperature,and disappearance of diarrhea symptoms.The self-healing group of mice had dull hair,high rectal temperature,stool with high water content,and the stool did not hold shapes when squeezed.Compared with the normal group,the mucosal structure of the duodenum,jejunum,and ileum in the self-healing group was partially damaged,with atrophy of the intestinal glands and inflammatory symptoms.Compared with the self-healing group,the mucosal structure in the treatment group was clearer,with tightly arranged intestinal epithelial cells and lamina propria cells,and the inflammatory condition was significantly improved.In comparison to the normal group,the CD of the duodenum in the self-healing group was higher(P<0.01),and the V/C and the number of goblet cells were lower(P<0.05,P<0.01)in the duodenum;the V/C in the treatment group was lower(P<0.05).Compared with the normal group,the VH and V/C of the jejunum and ileum in the self-healing group were significantly reduced(P<0.05,P<0.01).Compared with the self-healing group,the CD of the duodenum in the treatment group was significantly lower(P<0.01),while the V/C of the duodenum and ileum were significantly higher(P<0.05,P<0.01).In the duodenal segment,compared with the normal group,the AOD values of Claudin 1,Occludin,and ZO-1 proteins were lower in both the self-healing and treatment groups(P<0.05,P<0.01);compared with the self-healing group,the AOD value of Claudin 1,Occludin,and ZO-1 proteins in the treatment group were significantly higher(P<0.01).Compared with the normal group,the AOD value of Occludin protein in the jejunum and ileum was significantly lower in the self-healing group(P<0.01).Compared with the self-healing group,the AOD value of Claudin 1 protein in the jejunum was higher in the treatment group(P<0.05),and the AOD value of Occludin protein in the ileum was significantly elevated(P<0.01).Conclusion Gegen Qinlian Decoction can improve the level of intestinal inflammation in diarrhea mice with intestinal dampness-heat pattern and promote the expression of tight junction proteins Claudin 1,Occludin,and ZO-1.Its mechanism may be related to its repairing function on intestinal mechanical barrier.