| Objective: To observe the effects of self-made "Pingchang Decoction" on intestinal mucosal barrier function,tight junction protein and inflammatory factors in rats with ulcerative colitis(UC),the curative effect of Pingchang Decoction on UC rats was evaluated,and the mechanism of Pingchang Decoction in treating UC was discussed from the perspective of intestinal mucosal mechanical barrier and inflammatory response.Methods: 62 clean SD rats with half male and half female were selected as experimental subjects,and they were randomly divided into blank control group(n = 10)and experimental group(n = 52).Rats in the experimental group were given(TNBS)/ ethanol to establish the UC model.24 hours after the establishment of the model,2 rats in the experimental group were randomly killed,and the establishment of UC rat model was evaluated by observing intestinal mucosal injury and pathological manifestations.After successful induction of UC model,50 rats in the experimental group were again randomly divided into positive drug group(5-ASA),model group(Model),high-dose Pingchang Decoction group(PCT-H),middle-dose Pingchang Decoction group(PCT-M)and low-dose Pingchang Decoction group(PCT-L),each with 10 rats.Except for the normal feeding of the blank control group,all groups were treated by intragastric administration for 14 days.The model group was given0.9% sodium chloride solution,each dose group of Pingchang Decoction was given Pingchang Decoction according to different concentrations,and the positive drug group was given mesalachin suspension.The general conditions of rats were observed(including drinking water,food intake,stool,mental state,activity,body mass changes).After 2 weeks of intervention,the samples were taken,and the DAI was evaluated.The concentrations of IL-6,DAO and TNF-α in serum of rats were tested by ELISA.Colon tissues were stained with HE.The pathological manifestations of intestinal mucosa were observed under microscope,and the expression of occludin in intestinal tissue was measured by Western blot method.Results: 1.General situation: compared with the blank control group,the overall state of the rats in the initial stage of modeling was poor,showing listlessness,curled up into groups,lethargy,less movement,dull coat color,decreased food and water intake,mucous stool or watery stool,a small amount of bloody stool visible to the naked eye,increased stool frequency,obvious odor,dirty and moist cushion,and obvious fecal contamination around the anus.The body weight of rats decreased at the initial stage of the model,but increased in the later stage,but the range was smaller than that of the blank control group,with statistical difference(P < 0.05).After intervention,except that there was no obvious remission in the model group,the general condition of rats in the positive drug group and each dose group of Pingchang Decoction were improved in varying degrees compared with that before intervention.Among them,the overall improvement of rats in the high-dose group of Pingchang Decoction was the best,and the degree of improvement in the low-dose group was the lowest.There was no significant difference in body mass between the middle-dose of Pingchang Decoction and the high-dose of Pingchang Decoction(P > 0.05).Except 1 rat in the model group died due to improper operation on the 3rd day of intervention,there was no death in other groups.2.DAI score: after 14 days of intervention,compared with the blank control group,the DAI scores of rats in each group were significantly different(P < 0.05).Compared with the model group,there were significant differences in DAI scores among positive drug group,Pingchang Decoction middle-dose group,high-dose group and low-dose group(P < 0.05).Compared with the positive drug group,the DAI score of Pingchang Decoction low-dose group was higher,and the DAI score of high-dose Pingchang Decoction group was lower.There was no significant difference in DAI score between Pingchang Decoction middle-dose group and positive drug group(P > 0.05).3.Detection of rats serum DAO,TNF-α,IL-6 : after treatment,the concentrations of serum DAO,TNF-α and IL-6 in each group were significantly higher than those in the blank control group(P < 0.05).The concentrations of DAO,TNF-α and IL-6 in the positive drug group and Pingchang Decoction groups were significantly lower than those in the model group(P < 0.05).Among them,the concentration of DAO,TNF-α and IL-6 in the positive medicine group was higher than that in the middle and high dose group of Pingchang Decoction,but lower than that in the low dose group of Pingchang Decoction.The concentration of DAO,TNF-α and IL-6 in the high dose group of Pingchang Decoction decreased most obviously,with statistical significance(P < 0.05).4.The expression of Occludin protein: the expression level of occludin in colon tissue of rats in each group was significantly lower than that in the blank control group(P < 0.05).After intervention treatment,compared with the model group,the expression of occludin in the positive drug group and Pingchang Decoction each dose group was significantly higher than that in the model group(P < 0.05).Among them,the up-regulation was the most significant in the high-dose Pingchang Decoction group and the least in the low-dose Pingchang Decoction group(P < 0.05).5.Colon histopathology:compared with the blank control group,the intestinal mucosal epithelium of the model group was discontinuous,the crypt structure was destroyed,the defect was obvious,the local ulcer was deep to the muscular tissue,the glands were seriously injured,the arrangement was disordered,and inflammatory cell infiltration could be seen.In the positive drug group,the intestinal mucosal epithelium was incomplete,the crypt was moderately defective,the arrangement was irregular,and a small amount of inflammatory cell infiltration could be seen.In the high-dose of Pingchang Decoction group,the intestinal mucosal epithelium was continuous,the crypt structure was basically intact,and there was no obvious inflammatory cell infiltration.In the middle-dose of Pingchang Decoction group,the intestinal mucosal epithelium was incomplete,the crypt structure was slightly damaged,the arrangement was irregular,and a little inflammatory cell infiltration was found in the submucosa.In the low-dose Pingchang Decoction group,the intestinal mucosal epithelium was incomplete,the submucosal tissue was destroyed,and the crypt structure was moderately defective,irregularly arranged,with inflammatory cell infiltration.Conclusion: 1.Pingchang Decoction can improve the general condition,DAI score,colonic histopathological inflammation of UC rats,reduce the concentrations of serum TNF-α,IL-6 and DAO,and up-regulate the expression of occludin protein in colon tissue of rats.Pingchang Decoction can effectively treat UC rats,among which the high-dose of Pingchang Decoction has the best effect.2.This study confirmed that Pingchang Decoction can can repair TJ,improve the permeability of the intestinal mucosa,enhance the mechanical barrier function of the intestinal mucosa,and at the same time down-regulate the expression of pro-inflammatory cytokines,inhibit inflammation response,promote mucosal healing,thus playing a role in the treatment of UC model rats.3.The results of this experiment show that the self-made "Pingchang Decoction" has a definite curative effect on UC rats.Through the animal experiment,it has been confirmed again that "damp-heat of large intestine,weakness of spleen and stomach,liver depression and spleen deficiency,intestinal vein injury" are the etiology and pathogenesis of UC.“clearing heat and promoting dampness and detoxification,tonifying qi and nourishing blood and softening liver,cooling blood and astringent hemostasis,and relieving pain" are its treatment methods,which are in line with clinical practice and available for clinical selection. |
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