Study On The Treatment Of Ulcerative Colitis Damp Heat Syndrome Model Mice By Promoting Nlrp6 Inflammatory Body With Qingchang Huashi Recipe

Objective:To investigate the effect of Qingchang Huashi Recipe the expression of NLR family,pyrin domain containing 6(NLRP6)、related inflammatory factors and tight junction protein claudin-2、claudin-5 in Ulcerative Colitis mice with Dampness heat Syndrome.Methods:Sixty male C57BL/6 mice were randomly divided into control group,model group,Wetern medicine group(Mesalazine solution,0.1675g/kg/d)and low,medium and high-dose Chinese medicine group(8.125 g/kg/d、16.25g/kg/d、32.5g/kg/d),with 10 mice in each group.Except for the control group,the remaining groups were given distilled water containing 2.5%Dextran Sodium Sulfate for 1 week,combined with damp-heat type diet and environmental composite method to establish UC mice with dampness heat syndrome model.The drug was administered once a day for 7 days.Corresponding medication was respectively administered to each treatment group by gastrogavage,The control group and the model group were administered with the same doses of saline.The mice’s general physical condition,body weight,fecal traits,occult blood and disease activity index(DAI),the dejection amounts and weights in five hours were recorded daily.After the end of treatment.The length of colon was measured,the colonic pathological morphology was observed with Hematoxylin-eosin(HE)staining.Western blot was used to detect the colon tissue of mice NLR family,pyrin domain containing 6(NLRP6),caspase-1,claudin-2 and claudin-5 protein expression levels.Enzyme linked immunosorbent assay(ELISA)kit was used to detect the serum levels of interleukin-1β(IL-1β)in mice.The expressions of the genes of domain like receptor 6(NLPR6),caspase-1,claudin-2 and claudin-5 in the colon were detected by real-time PCR.Results Compared with the control group,the mice in the model group DAI increased(P<0.01)and had shorter colonic length(P<0.01);the pathological damage of colonic mucosa was severe,Results of Western blot showed that the expression of NLRP6、claudin-5 were down-regulated(P<0.05),the expression of caspase-1、claudin-2 were up-regulated(P<0.05,P<0.01).the serum IL-1β concentration was increased(P<0.05),Results of Real-time PCR indicated that the expression of NLRP6、claudin-5 decreased(P<0.01),the expression of caspase-1、claudin-2 increased(P<0.05、P<0.01).Compared with the model group,the mice in the western medicine group and the medium dose Chinese medicine group DAI decreased(P<0.01)and had longer colonic length(P<0.01);pathological conditions in the western medicine group and the medium dose Chinese medicine group were improved.Results of Western blot showed that the expression of NLRP6、claudin-5 in the medium dose Chinese medicine group were up-regulated(P<0.05),the expression of caspase-1 in the western medicine group and low、medium、high dose Chinese medicine group were down-regulated(P<0.05),the expression of claudin-2 in the western medicine group and medium、high dose Chinese medicine group were down-regulated(P<0.01);The serum IL-1β concentration in the western medicine group and low、medium dose Chinese medicine group were decreased(P<0.05、P<0.01、P<0.01).Results of Real-time PCR indicated that the expression of NLRP6 in the western medicine group and high dose Chinese medicine group were up-regulated(P<0.01、P<0.05),the expression of caspase-1 in the western medicine group and low、medium dose Chinese medicine group were decreased(P<0.05),the expression of claudin-2 in the western medicine group and medium、high dose Chinese medicine group were down-regulated(P<0.05),and the expression of claudin-5 in the medium、high dose Chinese medicine group were up-regulated(P<0.05).Conclusion:The UC mice model with dampness heat syndrome were successfully modeled,Qingchang Huashi Recipe has effect in treating UC mice with dampness heat syndrome,which may be related to its regulation of NLRP6 protein,related inflammatory factors and tight junction protein claudin-2、claudin-5,so as to reduce intestinal inflammatory reaction and relieve intestinal mucosal injury.