| Objective: To explore the possible role of miR-146 a in the pathogenesis of primary gout arthritis.Methods: 1.The correlation of mi R-146 a single nucleotide polymorphisms(SNPs),which including rs57095329,rs2910164,and rs2431697,with the susceptibility of primary gout:(1)A total of 606 cases of primary gouty arthritis(pGA)patients and 524 cases of healthy subjects(HC)were participanted in this study.Clinical and laboratory data were collected carefully.Blood samples were obtained from all of the participants.(2)The genotypes of miR-146 a SNPs were calculated using TaqMan probes that specially target the alternate alleles.(3)Statistical analysis: SPSS17.0software and SHEsis statistical software were used for analysis.Hardy-Weinberg test and genetic balance of genotype and allele frequencies distribution were tested by ?2 test.T test was used for comparison between groups.2.Changes of miR-146 a in different stages of patients with primary gouty arthritis:(1)The subjects were divided into 4 groups: 55 acute gout patients(AG),42 intercritical gout patients(IG),23 chronic gout patients(CG)and 43 healthy controls(HC).The expression of miR-146 a in peripheral blood mononuclear cells(PBMCs)was detected using real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)(TaqMan probe)in AG,IG,CG and HC groups.Clinical and laboratory parameters were collected in all participants.(2)The PBMCs of HC were stimulated with MSU(100ug/ml)for12h,and then measured IL-1? level,mi R-146 a,IRAK1 and TRAF6 mRNAusing ELISA and RT-qPCR respectively.(3)Statistical analysis: SPSS17.0software was used for analysis.Normal distribution measurement data was compared with t test,Non normal distribution measurement data was compared with the Wilcoxon rank test.The correlation between variables was analysed by Spearman correlation analysis.Recults: 1.The correlation of miR-146 a SNPs with the susceptibility of primary gout:(1)All 3 SNPs of miR-146 a followed Hardy-Weinberg equilibrium in participants(P>0.05).(2)The frequencies of AA,AG and GG genotypes of rs57095329 in p GA group were 64.7%,31.0%,and 4.3%,while the frequencies of AA,AG and GG genotypes of rs57095329 in controls were58.4%,35.9%,and 5.7%,respectively.There was no remarkably statistical difference in genotype frequencies between the two groups(P>0.05).The frequencies of A and G alleles of rs57095329 in pGA group were 80.2%,19.8%,and the frequencies of A and G alleles of rs57095329 in controls were76.3%,23.7%,respectively.There was significantly statistical difference in alleles frequencies between the two groups(P<0.05).Allele A was the risk factor for gout [OR(95%CI):1.261(1.124-1.414)].There was no significantly variance in distribution frequencies of genotypes and alleles in the other 2SNPs between the two groups.(2)The frequencies of AA,AG and GG genotypes of rs57095329 in GT group were 82.9%,15.8%,1.3%.The frequencies of A and G alleles of rs57095329 in GT group were 90.8%?9.2%,respectively.Significant differences were observed between the GT and HC groups with respect to genotype and allele frequencies of miR-146 a gene rs57095329(?2=16.631,P <0.01;?2=15.987,P <0.01).There was no significantly variance in distribution frequencies of genotypes and alleles in the other 2 SNPs between the GT and HC groups.(3)The frequencies of AA,AG and GG genotypes of rs57095329 in NTG group were 60.8%,34.4%,4.8%.The frequencies of A and G alleles of rs57095329 in NTG group were78.0%,22.0%,respectively.Significant differences were observed between the GT and NTG groups with respect to genotype and allele frequencies of miR-146 a gene rs57095329(?2=13.210,P<0.05;?2=12.646,P<0.01).(4)Significant differences were not observed between the NTG and HC groups with respect to genotype and allele frequencies of miR-146 a genes rs57095329,rs2910164 and rs2431697.2.Expression of miR-146 a in different stages of patients with primary gouty arthritis:(1)The expression of miR-146 a in PBMCs was significantly different in four groups(F=12.297,P<0.01,respectively),and the level of miR-146 a was significantly decreased in AG than that in IG and HC groups(0.1638±0.4310 vs 0.2631±0.5528,Z=-1.966,P<0.05;0.1638±0.4310 vs 0.2900±0.3631,Z=-3.308,P<0.01,respectively);the standard of mi R-146 a was remarkably reduced in CG when compared with HC group(0.2475±0.2350 vs 0.2900±0.3631,Z=-2.105,P<0.05,respectively);there were no significant difference among IG,CG and HC groups(P>0.05,respectively),and it was also no variance between AG and CG groups(P>0.05,respectively).(2)After stimulated with MSU in the PBMCs of HC,the expression of interleukin-1? was much higher than that in the control(92.63±4.70pg/ml vs 20.66±4.74 pg/ml;t=10.79,P <0.01,respectively),while the expression of miR-146 a was much lower than that in the control(1.19±0.09 vs 2.39±0.177;t=6.06,P <0.01,respectively),TRAF6 mRNA was significantly increased than that in the control(0.121±0.011 vs0.024±0.002;t=8.744,P <0.01,respectively),significant difference was not observed with respect to IRAK1 mRNA level between the twogroups(0.090±0.005 vs 0.079±0.005;t=1.539,P=0.162).(3)In AG,the miR-146 a level was negatively correlated with ESR(r=?0.355,P=0.031);and there was no significant correlation between the level of miR-146 a and another laboratory datas(P>0.05,respectively).(4)There was no significant difference in the expression of mi R-146 a in PBMCs of gout patients with genotypes of AG,GG,and AA in rs57095329.Conclusion:(1)The polymorphism of rs57095329 in miR-146 a may be associated with the susceptibility of primary gout in Chinese Han population.A allele may be a risk factor for gout.(2)The polymorphism of rs57095329 in miR-146 a may be associated with the susceptibility of chronic gout in Chinese Han population,and the A allele may be a risk factor for the pathogenesis of chronic gout.(3)The present data suggest that the polymorphism of rs2910164 and rs2431697 in miR-146 a is not associated with the occurrence of primary gout in Chinese Han population.(4)The changes of miR-146 a and its target molecule TRAF6 in MSU mediated gout inflammation suggest that mi R-146 a may be involved in the negative feedback regulation of gout inflammation and participate in spontaneous remission of gout inflammation. |
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