| Objective: to analyze the expression level of miR-223,miR-7 and NLRP3 splicing isomer 3(NLRP3-3),splicing isomer 4(NLRP3-4),NLRP3 mRNA in the peripheral blood mononuclear cells(PBMCs)of patients with primary gouty arthritis,Explore the relationship between the expression of miR-223,miRNA-7 and NLRP3 splicing isomers,and discuss the potential regulatory mechanisms.Methods: 1)The patients were divided into 4 groups:35 acute gout patients(AG),50 episodes gout patients(IG),25 chronic gout patients(CG)and 39 healthy patients,collecting their peripheral blood and laboratory indexes,2)detecting the expression level of mi R-223 and miR-7 in peripheral blood PBMCs of the four groups by real-time quantitative PCR(probe).3)detecting the expression level of NLRP3-3,NLRP3-4,NLRP3 in peripheral blood PBMCs of the four groups by real-time quantitative PCR(dye),Another 5 healthy volunteers were collected,stimulate their Peripheral blood with 100 μg/ml urate crystal(MSU)for 1 hour,the phosphate buffer(PBS)as control group,then detect the expression of miR-223、miR-7 and NLRP3-3,NLRP3-4,NLRP3、IL-1β by RT-qPCR.All data were analyzed by SPSS 19.0 software.Wilcoxon rank sum test and Spearman correlation analysis were used.Results: 1)the expression of miR-223 in AG,IG and CG groups was significantly higher than that in the healthy control group(p<0.05).In AG andIG,the expression of mi R-223 was significantly higher than that in CG,but no statistical difference was found between AG and IG(p>0.05).the expression of miR-7 in the four groups was statistically different(F=9.87,p<0.01).in the group of AG,IG and CG,The expression of miR-7 was significantly higher than in the healthy control group(p<0.05),but no statistical difference was found among the three groups(p>0.05).2)there was no statistical difference in the expression of NLRP3-3 in the four groups(F=1.752,p>0.05).in the group of AG,IG and CG,the expression of NLRP3-4 was significantly lower than that in the healthy control group(p<0.05),and the expression of NLRP3-4 in AG was significantly higher than that in CG,while no significant difference between AG and IG,in the group of AG,IG and CG,the expression of NLRP3 was significantly lower than that in the healthy control group(p<0.05),while no statistical difference among the three groups(p>0.05).3)by Spearman correlation analysis,in the group of CG,the expression level of miR-223 was positively correlated with NLRP3-3,NLRP3-4,NLRP3(r =0.558,0.564,0.537,p<0.05),while in AG,IG or healthy controls groups,no similar correlation was found(p>0.05);In AG,IG,CG or healthy controls,no significant correlation was found between miR-7 and NLRP3-3,NLRP3-4,NLRP3(p>0.05).After stimulating the peripheral blood of the health examiners by 100 μg/ml MSU for 1 hour,The expression level of miR-223 in the MSU stimulation group was significantly higher than that in the PBS control group(p<0.05),However,there was no statistical difference in the expression of mi R-7 between the MSU stimulation group and the PBS control group(p>0.05);The expression of IL-1β in MSU stimulation group was significantly higher than that in PBS control group(Z=-2.203,p<0.05),the expression of NLRP3-4 in MSU stimulation group was significantly lowerthan that in PBS control group(Z=-2.203,p<0.05),while NLRP3-3 and NLRP3 showed no significant difference between MSU stimulation group and PBS control group(p>0.05).5)In group AG,the expression of NLRP3 was positively correlated with UR(r=0.821,p=0.023).In group IG,the expression of miR-223 was positively correlated with RBC(r=-0.609,p=0.021).In group CG,the expression of NLRP3-3 and NLRP3 was positively correlated with ALB(r=0.886,0.829,p=0.019,0.042),and the expression of NLRP3-4 was negatively correlated with RBC(r=-0.786,p=0.036).In healthy control group,the expression of NLRP3-3,NLRP3 and miR-223 were negatively correlated with ALT(r=-0.634,-0.646,-0.706,p=0.049,0.044,0.034).Conclusion: 1)The expression of NLRP3 mRNA plays an important role in gouty arthritis,and NLRP3-4 may play a more important role than NLRP3-3 in gout,This effect may be related to the differential regulation of miR-223 and mi R-7 on NLRP3-3 and NLRP3-4.Overexpression of miR-223 significantly reduces NLRP3-4 expression,which negatively regulates gout inflammatory response.2)the expression level of miR-223 and NLRP3-3,NLRP3-4,NLRP3 in peripheral blood mononuclear cells of chronic gout patients may be in a state of stalemate.3)normal liver function may play an important role in maintaining the balance of inflammatory response in patients with gout,and the abnormal inflammatory reaction will affect the metabolism and blood cell stability. |
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