Effects Of Jiuxieling Granules On The Expression Of MyD88 And IRAK1 And Inflammatory Cytokines On Ulcerative Colitis Model Rats Of The Spleen And Kidney Yang Deficiency

ObjectiveTo observe the therapeutic effect of the treatment of UC in rats with deficiency of spleen and kidney yang,the effects of changes and pathological morphology of the colon tissue IL-2,IL-4,IL-10 and inflammatory factors of TLR signaling pathway related genes MyD88,IRAK1 protein expression,explained taking Jiuxieling granules in the treatment of UC curative effect,mechanism and related therapeutic targets.Methods90 SPF Wistar rats were randomly divided into blank group,model,using rhubarb decoction plus hydrocortisone +TNBS/ ethanol enema method to establish animal model.After successful modeling,the model group was randomly divided into three groups: UC model group of spleen and kidney yang deficiency,high,medium and low dose group and positive control group(group SASP).Based on the gross morphology and colon tissue were observed under microscope after HE staining were detected by ELISA in rat serum IL-2,IL-4,IL-10 and RT-qPCR were used to detect the content of rat colon tissue MyD88,IRAK1 gene expression,immunohistochemistry and Western blotting analysis of rat colon tissue,MyD88 the expression of IRAK1 protein.Results1.The content of 1 ELISA for the detection of serum IL-2,IL-4 and IL-10: compared with the blank group,reduce the content of rat serum in model group IL-2,IL-4,IL-10(P<0.01),the difference was statistically significant;compared with the model group,Liang Shenggao group of rat serum containing IL-2,IL-4 and IL-10 of each treatment(P<0.05),the difference was statistically significant,most notably Jiuxieling granules in high dose group(P<0.01).2.The relative expression of colon tissue were detected by RT-qPCR in 2 MyD88 and IRAK1 mRNA: compared with the model group,the relative expression decreased significantly in colon blank group rats MyD88,IRAK1 m RNA,had significant difference(P<0.01);compared with the model group,the relative expression in colon tissue of rats ineach treatment group in MyD88,IRAK1 mRNA the amount also reduced(P<0.01),and decreased in high dose group compared with the most;Jiuxieling granules in high dose group,low dose group,SASP dose,with obvious difference between groups of Jiuxieling granules,with statistical significance(P<0.05).3.Immunohistochemical method was used to detect the expression of MyD88 and IRAK1 protein in the colon tissue of rats.The expression of MyD88 and IRAK1 in the colon of rats were observed under the microscope.Compared with the blank group,the positive expression of MyD88 and IRAK1 were in the colon of rats in the model group,better than the blank group,the average optical density value increased significantly(P<0.01),the difference was significant;compared with the model group,each treatment group rat colon MyD88,IRAK1 positive expression was weak or moderate expression,is still strong in the the blank group,the average optical density value decreased(P<0.01),the most significant decline is the high dose group,significantly better than the SASP group;compared Jiuxieling granules in high dose group,low dose group,SASP dose,with obvious difference between groups of Jiuxieling granules,with statistical significance(P<0.05).4.The expression of MyD88 and IRAK1 protein in colon tissue were detected by Western blotting: compared to the blank group,MyD88 increased,IRAK1 protein expression in colon tissue of the rats in the model group,the difference was significant(P<0.01);compared with the model group,the colon tissue of rats in each treatment group the expression of MyD88 and IRAK1 protein decreased significantly(P<0.01),especially in high dose group significantly;compared Jiuxieling granules in high dose group,with statistical significance of Jiuxieling granule and low dose SASP group,the differences between groups(P<0.05).Conclusion1.The contents of IL-2,IL-4 and IL-10 were increased,and the inflammatory factors could inhibit the infiltration of inflammatory cells,reduce the inflammation of mucosa,improve the injury of tissue,and promote the repair of colonic mucosa.2.Spleen kidney yang deficiency type UC disease and MyD88,IRAK1 gene and protein level has some connection,indicating that MyD88,IRAK1 expression of cocoa as evaluation index,determine the severity of UC disease.3.Related to down-regulation of Jiuxieling granule in treating UC disease may be relatedto MyD88,IRAK1 gene and protein,and inflammation,repair of inflammatory injury of intestinal mucosa,the evaluation index can be used as a clinical treatment effect of UC.