| Objective: To explore the therapeutic effect of Kuijie prescription on ulcerative colitis model rats and the mechanism of Th17/Treg immune balanceMaterials and Methods: Healthy male rats were randomly divided into blank group,model group,Kuijie prescription group and sulfoazine group,with 8 rats in each group.Except the blank group,the UC rat model was established by TNBs-ethanol method in the other 3 groups.Rats in blank group and model group were intragastrically given purified water,Kuijie prescription group was intragastrically given Kuijie prescription,and salazine sulapyridine solution was intragastrically given for 14 consecutive days.The general condition of rats was recorded daily and DAI score was performed.CMDI score was performed on colon tissue. Colon tissue was stained with HE and TDI score was performed.The contents of IL-6,IL-17,IL-10 and TGF-β1 in serum were detected by ELISA.The expressions of IL-6,IL-17,IL-10 and TGF-β1 in colon tissues were detected by immunohistochemistry.The expression of ROR-γt and Foxp3 m RNA in colon tissues was detected by RT-q PCR.Results:1.General conditions and DAI score: After modeling,rats showed sluggish response,decreased activity,dark fur,significantly decreased food intake and water intake,bloodstained runny stool,and foul perianal odor.After 14 days of intragastric treatment,DAI score in model group was significantly higher than that in blank group(P < 0.01).DAI scores of Kuijie prescription group and salazine group were lower than those of model group(P < 0.01).2.CMDI score: After 14 days of intragastric treatment,colon tissue of rats in blank group was normal without adhesion to the surrounding area;Colonic tissue hyperemia,edema,erosion and ulceration of rats in model group had a wide range of lesions,and colonic tissue was dark in color and adhered to surrounding tissues.CMDI score of rats in model group was significantly higher than that in blank group(P < 0.01).The surface of colon tissue in Kuijie prescription group and salazine group was not smooth,there was a small amount of congestion,edema and erosion,the lesions were small,there was no adhesion with surrounding tissues,CMDI score was lower than that in model group(P < 0.01).3.HE staining: After 14 days of intragastric treatment,the overall structure of colon tissue of rats in blank group was intact without obvious inflammatory cell infiltration;There were inflammatory cell infiltration,goblet cell deformation or disappearance,crypt abscess,and TDI score in model group was significantly higher than that in blank group(P < 0.01).Kuijie prescription group and salazine group were significantly improved,inflammatory infiltration was not obvious,crypt structure was basically complete and orderly,and TDI score was lower than that of model group(P < 0.01).Compared with salazine group,colonic tissue of Kuijie prescription group was recovered better,which was close to normal rats(P < 0.05).4.ELISA results: After 14 days of intragastric treatment,the contents of IL-6 and IL-17 in serum of rats in two treatment groups were lower than those in model group(P < 0.01),while the contents of IL-10 and TGF-β1 were higher than those in model group(P < 0.01). Compared with salazine group,the contents of IL-6 and IL-17 in serum of Kuijie prescription group were decreased(P < 0.05),while the contents of IL-10 and TGF-β1 were increased(P <0.05).5.Immunohistochemical test results: After 14 days of intragastric treatment,the positive expressions of IL-6 and IL-17 in colon tissue of rats in the two treatment groups were lower than those in the model group(P < 0.01),while the positive expressions of IL-10 and TGF-β1were higher than those in the model group(P < 0.01).Compared with salazine group,the positive expressions of IL-6 and IL-17 in colon tissues of Kuijie prescription group were decreased(P < 0.05),while the positive expressions of IL-10 and TGF-β1 were increased(P <0.05).6.RT-qPCR results: After 14 days of intragastric treatment,the expression of Foxp3 m RNA in the colon tissues of rats in the two treatment groups was higher than that in the model group(P < 0.01),and the expression of ROR-γt m RNA was lower than that in the model group(P <0.01).Compared with salazine group,the expression of Foxp3 m RNA in Kuijie prescription group was increased(P < 0.05),and the expression of RER-γt m RNA was decreased(P <0.05).Conclusion:1.Kuijie prescription can significantly improve the mental state,body weight,stool characteristics and blood in stool of ulcerative colitis model rats established by TNBs-ethanol method.2.Kuijie prescription can reduce CMDI and TDI scores of colonic tissue in ulcerative colitis model rats,and alleviate intestinal mucosal injury.3.Kuijie prescription has certain effects on ulcerative colitis model rats,and its mechanism may be through down-regulating the expressions of IL-6,IL-17 and ROR-γt and upregulating the expressions of IL-10,TGF-β1 and Foxp3,thus regulating the immune balance of Th17/Treg and maintaining intestinal immune homeostasis. |
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