| Objective To explore the effect of Gancao Xiexin decoction on the expression of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(My D88)and β-arrestin(inhibitory protein)in TLRs/NF-k B signal transduction pathway in mice with ulcerative colitis induced by sodium dextran sulfate((DSS)),and to explore the regulatory mechanism of Gancao Xiexin decoction on ulcerative colitis,so as to provide guidance for clinical treatment.Methods 60 SPF clean mice were randomly divided into blank group,model group,mesalazine group and licorice Xiexin decoction high and low dose group with 10 mice in each group.The other 5 groups except the blank group were given 100 ml free drink of 3%DSS solution,and the blank group was given free drink of 100 ml distilled water for 7 days.After the success of the model,the mesalazine group was given0.17 g / kg / day,the model group and blank group were given normal saline,and the high and low dose group of Glycyrrhiza Xiexin decoction was given 16.4 g/kg/d,8.2g/kg/d and 4.1g/kg/d ’s concentrated solution of Glycyrrhiza Xiexin decoction twice a day for 7 days.DAI was observed and recorded,After administration,the mice were killed by cervical dislocation method,the whole colon from anus to cecum was cut off,the length of colon was measured,the morphological changes of intestinal tissue and pathological score of mice in each group were observed by HE staining,and the expression of TLR4,My D88 and β-arrestin protein in colon tissue of mice in each group was detected by Western Blot method.Result 1.(DAI)Evaluation of disease activity index: there was no significant difference in general activity,body weight,coat color,diet,feces and so on in each group before modeling.After the establishment of the model,compared with the blank group,the intervention mice were worse in general activity,weight loss,dark coat color,reduced diet,thin stool,hematochezia and so on.After drug treatment,the above-mentioned state was significantly alleviated(P < 0.05).2.Evaluation of colon length: compared with the blank group,the colon length of the other five groups was shortened(P < 0.05),and compared with the model group,the colon length of mice in mesalazine group and high and low dose group of Glycyrrhiza Xiexin decoction increased(P < 0.05).Compared with the mesalazine group,the colon length of mice in the high and low dose group of licorice Xiexin decoction was shortened(P < 0.05).The intervention length of licorice Xiexin decoction was as follows: high dose group > middle dose group > low dose group.3.Pathological score of colon tissue:compared with the blank group,the pathological score of colon tissue of the other five groups increased(P < 0.05),and that of the four groups treated with drugs decreased(P < 0.05),compared with the model group.Compared with the mesalazine group,the pathological score of colon tissue in the high and low dose group of licorice Xiexin decoction increased(P < 0.05).The pathological score of colon tissue in the intervention of licorice Xiexin decoction was as follows: high dose group > middle dose group > low dose group.4.Western Blot detection method: compared with the blank group,the expression level of TLR4,My D88 and protein in colon tissue of model group increased,while the expression level of β-arrestin protein decreased,(P< 0.05).Compared with the model group,the expression level of TLR4 and My D88 protein decreased and the expression level of β-arrestin protein increased in colon tissue of mice treated with drugs,(P < 0.05).The protein expression levels of TLR4 and My D88 were as follows: mesalazine group < high-dose licorice Xiexin decoction group < middle-dose licorice Xiexin decoction group < low-dose licorice Xiexin decoction group;the expression level of β-arrestin protein was as follows: mesalazine group > high-dose licorice Xiexin decoction group > middle-dose licorice Xiexin decoction group > low-dose licorice Xiexin decoction group.Conclusion Gancao Xiexin decoction has obvious effect on ulcerative colitis,and its mechanism may be related to inhibiting the protein expression of pro-inflammatory factor TLR4 and My D88 in colon tissue and increasing the protein expression ofβ-arrestin in anti-inflammatory factor through TLRs/NF-k B signal transduction pathway,so as to reduce inflammatory reaction,regulate immune system,repair intestinal mucosa and maintain intestinal homeostasis,especially in high dose Gancao Xiexin decoction group.Therefore,licorice Xiexin decoction can provide guidance for clinical treatment of UC. |
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