| Objective: To observe the effect of Hua-yu-tong-yang Recipe on the content of Interleukin-23(IL-23)and JAK-STAT signaling pathway in rats with ulcerative colitis,so as to explore its mechanism of action.Methods: 32 healthy Wistar rats were divided into blank control group with 8 rats and model replication group with 24 rats,according to the method of random number table.The model rats with ulcerative colitis were established by gavage and free-drinking with 5% dextran sodium sulfate(DSS)solution.After 7 days,2 rats in blank control group and 6 rats in model replication group were killed at random.The colonic tissue of rats was dissected immediately.The success of the model was verified by observing the length of the intestine,mucosal ulcer and histopathological changes.On the second day after the successful establishment of the model,the rats in the model replication group were divided into Hua-yu-tong-yang group,mesalazine group and model group according to the method of random number table,six rats in each group.Each group was given corresponding drug enema intervention,Hua-yu tong-yang group was given Hua-yu-tong-yang Recipe(0.54g/ml),mesalazine group was given mesalazine enema solution(0.036g/ml),model group and blank control group were given saline enema.The dosage of each group was 2ml/animal/day,once a day,lasting for 10 days.DAI score was used to evaluate the general situation of rats during the experiment.At the end of the treatment,all rats were killed by cervical dislocation method,and the colon of rats was dissected immediately.The length of colons in each group was compared.CMDI score was used to evaluate the degree of colonic tissue damage in rats.HE staining was used to observe the pathological changes of colon tissue,the content of IL-23 in colon tissue was detected by ELISA,and the expression of JAK1,TYK2,STAT1 and STAT4 protein in colon tissue was detected by Western Blot.Results: the results of DAI score showed that: during the model-making period,the DAI score of the model group showed a significant upward trend compared with the blank control group;at the end of the treatment,the DAI scores of rats in Hua-yu-tong-yang group and mesalazine group were significantly lower than those in model group.The results of colon length in rats showed that: the colon length of rats in the model group was significantly shortened compared with the blank control group(P<0.01);while that in the Hua-yu-tong-yang group and mesalazine group showed significant improvement in colon tract shortening(P<0.01).The results of CMDI in colon tissue of rats showed that: the colonic mucosal surface of the model group was rough,the colonic wall was thickened,ulcers were formed,and mucosal damage was severe compared with the blank control group(P<0.01);the colon injury in Hua-yu-tong-yang group and mesalazine group was significantly improved compared with the model group(P<0.01),and there was no significant difference between the two groups(P>0.05).The results of HE staining and TDI score showed that: the colon tissue of the model group was the most severely damaged,and the structure of the glands in the lamina propria was damaged,the structure of the crypt was disordered,and the inflammatory cells infiltrated into the submucosa compared with the blank control group(P<0.01);the glands in the lamina propria of the colon mucosa of the Hua-yu-tong-yang group and the mesalazine group were basically arranged in order,and a small number of inflammatory cells were observed compared with the model group(P<0.01),and there was no significant difference between the two groups(P>0.05).The results of ELISA showed that: the content of IL-23 in colon tissue of model group was significantly higher than that of blank control group(P<0.01);and that of Hua-yu-tong-yang group and mesalazine group was lower than that of model group(P<0.01),and there was no significant difference between the two groups(P>0.05).Western Blot showed that: the expression of JAK1,STAT1 and STAT4 in colon tissue of model group was higher than those of blank control group;and the expression of JAK1 and STAT1 in colon tissue of Hua-yu-tong-yang group and mesalazine group decreased significantly(P<0.01),the expression of STAT4 decreased(P<0.05),while the change of TYK2 protein was not significant(P>0.05).Conclusion:1.The expression of IL-23 in colon tissue of UC rats was positively correlated with the expression of JAK1 and STAT4 protein,and the expression of JAK1 protein was positively correlated with the expression of STAT1 and STAT4 protein.It is speculated that the IL-23 /JAK-STAT signaling pathway may be a new way of pathogenesis of UC;2.Hua-yu-tong-yang Recipe may reduce the level of IL-23,regulate the JAK-STAT pathway,and reduce the expression of JAK1,STAT1 and STAT4 proteins,so as to reduce the colon inflammatory response and mucosal damage,and play a role in UC lesions. |
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