Effect Of Gancao Xiexin Decoction On TLR4/MyD88/NF-κB Inflammatory Signaling Pathway In Ulcerative Colitis Rats

Objective: Ulcerative colitis(UC)is a difficult disease in modern times.Traditional Chinese medicine(TCM)is effective.It is a hot topic in the study of digestive system diseases.By observing the general condition,DAI、TNF-α、IL-10 and TLR4、My D88、NF-κB p65 protein expression and gene transcription of UC rats,the mechanism of Gancao Xiexin Decoction on ulcerative colitis model rats was studied based on TLR4/My D88/NF-κB signaling pathway.Methods: 6-8 w male Wistar rats were randomly divided into blank group,model group,western medicine group and high,medium and low dose group.A model of ulcerative colitis induced by 2,4-dinitrochlorobenzene and acetic acid,gavage for 2 weeks.Observation of the general situation of rats and assessment of disease activity index(DAI);HE staining method was used to compare the morphology of colonic tissue in rats,To observe the effect of Gancao Xiexin Decoction on colon histomorphology of model rats;ELISA method was used to detect and compare the IL-10、TNF-α expression levels in peripheral blood of rats;Western Blot detection of TLR4、My D88、NF-κB p65 protein expression in rat colon;QPCR transcription of colon tissue TLR4 、 My D88 、 NF-κB p65 m RNA rats in each group was detected.To investigate the therapeutic effect of Gancao Xiexin Decoction on ulcerative colitis rats,And the mechanism of TLR4/My D88/NF-κB signaling pathway.Result:1.The general condition and DAI: the rats induced by 2,4-dinitrochlorobenzene combined with acetic acid showed weight,mental state,diet loss,sparse and dim hair,increased defecation,thin stool and blood stool,and the symptoms gradually increased during the modeling period.The symptoms improved gradually and the body weight increased gradually after stopping the modeling and gavage.The weight gain in the treatment group was lower than that in the blank group and higher than that in the model group,Chinese medicine high dose group DAI lower than western medicine group(All P<0.05).2.Colonic morphology observed by HE staining: Colonic mucosal epithelium,glandular arrangement and structure were intact in the blank group under light microscope 。 In the model group,the colonic mucosal epithelium was incomplete,the crypt atrophy and deformation,the disorder of arrangement,and a large number of inflammatory cells exudated and infiltrated.3.ELISA detected IL-10 and tumor necrosis factor TNF-α: with the lowest serum TNF-α concentration,low TNF-α in the treatment group,the highest IL-10 in the model group,and the serum orthostatic factor IL-10 in the treated group was higher than the model group,(All P<0.05).TNF-α in the high,medium and low-dose group showed the upward trend,the high,medium and low-dose group and the western medicine group IL-10 showed the downward trend,and TNF-α tested differences between the high-dose group and the western medicine group(P<0.05).4.Westorn blot method: the expression of TLR4、My D88、NF-κB p65 protein in colon tissue of blank group was the lowest,and the expression of TLR4、My D88、NF-κB p65 protein in colon tissue of model group was significantly higher than that in each administration group(P<0.05).Gancao Xiexin Decoction high,middle and low dose group and western medicine group rat colon tissue TLR4、My D88、NF-κB p65 protein content showed an upward trend,There was significant difference between high dose and western medicine group(P<0.05),TLR4 and NF-κB p65 protein,middle dose and western medicine group(P<0.05).5.QPCR detection of TLR4、My D88、NF-κB p65 m RNA transcription in rats:TLR4、My D88、NF-κB p65 m RNA transcription in colon tissue was lowest in blank group,and the TLR4、My D88、NF-κB p65 m RNA transcription of model group was significantly higher than in each administration group(All P<0.05).The TLR4 、My D88 、NF-κB p65 m RNA transcription of colon tissue in high,medium and low dose group and western medicine group showed the rise,with the difference between high dose group and western medicine group(P<0.05),and the NF-κB p65 m RNA difference in middle dose group and western medicine group(P<0.05).Conclusion:Gancao Xiexin Decoction has a remarkable effect on the treatment of UC rats.It can effectively improve the general situation and DAI,of UC rats,inhibit serum pro-inflammatory factors TNF-α,up-regulate anti-inflammatory factors IL-10,down-regulate TLR4、My D88、NF-κB p65 m RNA transcription,inhibit their protein expression.Effectively inhibit the activation of TLR4/My D88/NF-κB signaling pathway and ultimately inhibit ulcerative colitis,Gancao Xiexin Decoction is similar or better than sulfasalazine.Different doses of Gancao Xiexin Decoction can affect the curative effect.