| ObjectiveBased on the theory of Ulcerative Colitis(UC)and the relationship between UC and NLRP3 inflammasome/interleukin-1 beta/interleukin-17(NLRP3/IL-1?/IL-17)signaling pathway,we investigated the molecular mechanism of Five-Flavor Sophora flavescens Enteric-Coated Capsules(FSEC)-a national new medicine-on dextran sulphate sodium salt(DSS)induced UC mice by animal experiment.We explain the target and mechanism from the molecular biology level and provide theoretical basis for clinical application.Methods1.20 male BALB/c mice were randomly divided into normal control(Nctrl,n=10)group and model group(n=10).Model group drunk DSS freely for 7 days to make successfully mice model of UC,while Nctrl group drink distilled water freely.Observe their mental state,weight,stool consistency,faecal occult blood,colours and diseases activity index(DAI)so on after administration,then observe the pathological changes of colon tissues under light microscope to evaluate and analysis the mice model of UC.2.50 male BALB/c mice were randomly divided into five groups:Nctrl group,model group,low dose of FSEC group(1.92g/(kg · d)),middle dose of FSEC group(3.84g/(kg · d))and high dose of FSEC group(7.68g/(kg · d)),ten in each group.Drinking water with 3%DSS for seven days in all groups except Nctrl to make mice model of UC;Then Nctrl group and model group received 0.9%saline enema for seven days,while FSEC groups had FSEC suspension enema of different concentration respectively as mentioned before,one time each day for seven days.Observe their mental state,weight,stool consistency,faecal occult blood,colours and diseases activity index(DAI)so on after administration,then observe the pathological changes of colon tissues under light microscope to evaluate and analysis the mice model of UC.Detect the concentrations of caspase-1 interleukin-1 beta(IL-1?),retinoid-related orphan receptor gamma t(ROR y-t),interleukin-17(IL-17)and interleukin-6(IL-6)·3.30 male BALB/c mice were randomly divided into five groups:Nctrl group,model group,low dose of FSEC group(1.92g/(kg · d)),middle dose of FSEC group(3.84g/(kg · d))and high dose of FSEC group(7.68g/(kg · d)),six in each group.Drinking water with 3%DSS for seven days in all groups except Nctrl to make mice model of UC;Then Nctrl group and model group received 0.9%saline enema for seven days,while FSEC groups had FSEC suspension enema of different concentration respectively as mentioned before,one time each day for seven days.The mice were killed 24 hours after the last enema and fasted.The colon(1 cm above anus to 1 cm below the ileocecal end)was extracted to detect the mRNA expression of NLRP3,IL-1 ? in colonic tissue by quatity polymerase chain reaction(qPCR)method.Results1.Compared with Nctrl group,the mice in the model group were in poor mental state,like to curl up and push,the fur was dark,the weight was decreased,sticky and bloody stool was observed in the naked eye,the DAI score of the model group was higher than that of the normal control group;the pathological changes of the model group were observed under light microscope,In the normal control group,the colonic tissue structure was integrated,while in the model group,the colonic tissue structure was destroyed,the crypt and the goblet cell were decreased,the gland was destroyed,a large number of inflammatory granulocytes infiltrated,and local ulcers were formed.4.Compared with Nctrl group,the colonic length of other groups was shortened in varying degrees,the colonic length in the model group was significantly different from that in the middle and high dose of FSEC groups(P<0.01).Compared with Nctrl group,the spleen coefficient of other model groups was higher in different degree,and the colon coefficient of other model groups was lower in different degree.Compared with the model group,the colonic coefficients of FSEC groups were increased in varying degrees,and the middle and high dose groups decreased significantly(p<0.05).The colonic coefficients of FSEC groups were significantly higher(p<0.05).Compared with Nctrl group,the protein level of Caspase-1.IL-1?.ROR?-t?IL-17?IL-6 and the activity of MPO in colonic tissue of mice in model group increased(p<0.01),while FSEC groups reduced in different degree.Compared with model group,the protein level of Caspase-1?IL-1? ROR?-t?IL-17?IL-6 in the middle dose of FSEC group remarkly reduced(p<0.01),and the activity of MPO reduced(p<0.05)?5.Compared with Nctrl group,the mRNA level of NLRP3 and IL-1 3 in colonic tissue of mice in model group increased(p<0.01).while compared with model group,these mRNA level in FSEC groups reduced(p<0.05)?2.Conclusion1.The model of UC mice could be successfully induced by the method of free drinking of 3%DSS for 7 days.On the basis of this,the mechanism of FSEC treating UC could be further studied.2.The mechanism of FSEC treating UC mice was related to its inhibition of NLRP3/IL-1?/IL-17 signaling pathway... |
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