| Purpose:By establishing a mouse model of spleen deficiency STC and comparing the treatment effect of Simotang with that of positive drug control group,model group and blank group,the therapeutic effect of Simotang in the treatment of spleen deficiency STC was discussed,and the mechanism of action of Simotang was initially explored whether it was related to SP,VIP and ICC,providing theoretical basis and experimental basis for clinical application of Simotang in the treatment of STC.Material and method : Sixty clean grade Kunming mice were randomly divided into 2 groups,including 10 blank group and 50 model group.The model group was modeled by Zou Ying’s method of replicating spleen deficiency constipation model,that is,the mice were given decoction of senna leaf water for 7days to cause chronic diarrhea and spleen deficiency,and the mice were fed low-fiber rice for 7 days every other day to cause constipation.The seven days were the same time limit of food and water,hunger and satiety disorders to maintain spleen deficiency state,a total of 15 days.After successful replication of the model,mice in the model group were randomly divided into 5 groups: model group,Moxabili group,Simotang low-dose,medium-dose and high-dose groups,which were intragastric administration of corresponding drugs for 7 days,and blank group and model group were intragastric administration of equal doses of normal saline.Treatment ends,all mice fasting,the water after 24 hours out,first by gastric homemade ink 0.2 ml / 10 g,eyeball blood after 30 minutes,then put to death in mice and should be fixed,abdominal cut out complete intestinal tissue,record the data,finally shearing near the anus bowel tissue fixed in the group fixed liquid.The general behavior of mice,fecal moisture content and body weight before and after treatment were recorded.The ink advancement rate of mice in each group was compared.Hematoxylin-eosin staining was used to observe the morphological differences and pathological changes of colon tissue,the content changes of SP and VIP were analyzed by enzyme-related immunosorbent assay,and the protein expression of ICC was investigated by immunohistochemistry.Results:1.Molding result: After the modeling,compared with the blank group,the mice in the model group were small and shriveled,crouched under the cushion material,with dark and unglossy fur,reduced defecation,dry,hard and granular feces,and even some mice were black around the anal area,with significant weight loss(P < 0.01),significant decrease in fecal moisture content(P < 0.01),and significant decrease in ink propulsion rate(P < 0.01).In line with the clinical manifestations of spleen deficiency STC,the modeling was judged to be successful.2.Treatment results: After treatment,the body weight of Simotang(low,medium and high dose)groups increased significantly compared with model group(P < 0.05),the activity of mice increased,the coat gloss increased,and the defecation basically returned to normal,among which the difference was the most significant in Simotang high dose group(P < 0.01),and the fecal moisture content increased significantly(P <0.01).The advancing rate of ink was significantly increased(P < 0.05).3.Hematoxylin-eosin staining results: Compared with blank group,intestinal villi in model group were loose,goblet cells were reduced,glands were deformated and atrophied,submucosal interstitial edema was observed,obvious bleeding was observed,inflammatory cell infiltration was serious,and intestinal tissue pathological damage was the most serious.Compared with model group,a small amount of villi were broken and shed in intestinal tissue of mice in Simotang(low,medium and high dose)group and Moxabili group.The morphological damage of intestinal tissue recovered obviously,the inflammatory infiltration was light,and the pathological damage of intestinal tissue was significantly relieved.4.Elisa assay results: Compared with blank group,SP expression in model group was significantly decreased(P < 0.01),and VIP expression was significantly increased(P< 0.01).Compared with model group,SP was significantly increased in Simotang(low,medium and high dose)groups and Moxabili groups(P < 0.05),while VIP expression was significantly decreased(P < 0.05).The effect of Simotang high dose group was the best(P < 0.01).5.Immunohistochemical results: Compared with blank group,the expression of ICC marker c-kit in model group was significantly decreased(P < 0.05).After treatment,the expression of c-kit in Simotang(low,medium and high dose)groups and Moxabili group was significantly increased compared with model group(P< 0.05).Conclusion:1.Low,medium and high doses of Simotang can significantly improve the fecal moisture content and ink advancing rate of mice,and obviously improve the symptoms of spleen deficiency in mice.2.Simotang can effectively treat spleen deficiency STC,among which high-dose Simotang has the best therapeutic effect on spleen deficiency STC model mice.3.Simotang may achieve therapeutic purposes by up-regulating the expression of SP and ICC,down-regulating the expression of VIP,balancing the contraction and diastole of colon tissue,and repairing the pathological damage of colon tissue. |
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