Study On The Effect Of Warming The Kidney And Strengthening The Spleen On Gene And Protein Expressions Of The Negative Regulatory Factor Of SOCS2/3 In Ulcerative Colitis Model Rats’ Colon

Objective Ulcerative colitis(UC) is a kind of inflammatory bowel disease(IBD) with complex and diverse clinical manifestations.In recent years, the incidence of UC continues to grow, and has a high risk of cancer, but modern medicine is still lack of specificity of the treatment, so it brought a serious impact on patient and society.The present study shows that the incidence of UC is closely related to the imbalance of intestinal mucosal immune homeostasis by the excessive activation of JAK/STAT signaling pathway, so it is necessary to carry out a precision negative regulation of JAK/STAT signaling pathway and to avoid excessive immune response to restore the immune balance. From the perspective of negative regulatory factor,The topic choose negative regulators SOCS2, SOCS3 of JAK/STAT signaling pathway,to disscuss the mechanism of Sishen Pill on spleen-kidney yang deficiency of UC from the level of gene and protein.Method Using senna orally + hydrocortisone injection intraperitoneal injection + 2,4,6-trinitrobenzene sulfonic acid(TNBS) / ethanol enema to establish the rat model of spleen-kidney yang deficiency of ulcerative colitis. 90 Wistar rats were randomly divided into the blank group, model group, Salazosulfadimidine(SASP) group and the Sishen pill(high dose 、medium dose 、 low dose)group. After the models successfully, Sishen pill of high,medium and low dose group were given drug 10,5,2.5g/kg/d orally,the SASP group was given SASP suspension 0.36g/kg/d orally, the model group was treated with equal amount of normal salinefor 21 days. To observe the morphology and colon injury of rats and the pathological changes of colon in rats, the method of RT-q PCR was used to detect the expression of SOCS gene, SP immunohistochemistry and Western blot were used to detect the expression of SOCS3 and SOCS2 in colon of rats.Result1.The result of RT-q PCR:SOCS:Compared with the blank group, the content of SOCS m RNA in the model group was decreased(P<0.01);Compared with the model group,the content of SOCS m RNA in the treatment groups were increased(P<0.01,P<0.05).2.The result of SP:(1)SOCS2 1Positive cells in the blank group, has a high expression in the mucosal layer and submucosal layer of the colon tissue of the rats,showed brown particles.The model group was low expression.The treatment groups showed a high or moderate expression.2The result of Average optical density value:Compared with the blank group, the expression of SOCS2 in the model group was decreased(P<0.01);Compared with the model group,the expression in the treatment groups were increased(P<0.01).(2)SOCS31Positive cells in the blank group, has a high expression in the mucosal layer and muscular layer of the colon tissue of the rats, showed brown particles.The model group was low expression.The treatment groups showed a high or moderate expression.2The result of Average optical density value:Compared with the blank group, the expression of SOCS3 in the model group was decreased(P<0.01);Compared with the model group,the expression in the treatment groups were increased(P<0.01,P<0.05).3.The result of WB:(1)SOCS2 :Compared with the blank group, the expression of SOCS2 in the model group was decreased(P<0.01);Compared with the model group,the expression in the treatment groups were increased(P<0.01,P<0.05).(2)SOCS3 :Compared with the blank group, the expression of SOCS3 in the model group was decreased(P<0.01);Compared with the model group,the expression in the treatment groups were increased(P<0.01).Conclusion1.It can successfully establish the rat model of spleen-kidney yang deficiency deficiency of ulcerative colitis by using senna orally + hydrocortisone injection intraperitoneal injection+ 2,4,6- trinitrobenzene sulfonic acid(TNBS) / ethanol enema2.Spleen-kidney yang deficiency is an important pathogenesis of UC,Syndromes of Spleen-kidney yang deficiency is an important syndrome UC,warming the kidney and strengthening the spleen is an important threatment of UC.3.Sishen Pill can increase the SOCS m RNA and the protein content of SOCS2, SOCS3 in the colon tissue of the rats with Spleen-kidney yang deficiency of UC,thereby inhibiting inflammatory response and recovery of immune homeostasis, to achieve the purpose of the treatment of UC.4.The mechanisms of Sishen Pill in treatment of UC may be interfered with the expressions of negative regulatory factors SOCS2/3 of JAK/STAT signaling pathway in the microenvironment of UC.To regulate and control the expressions of negative regulatory factors may be an important targets for the treatment of UC.