| Objective: This study aims to investigate the expression and clinical significance of Phosphofructokinase 1(PFK1)in the peripheral blood of patients with primary gouty arthritis(GA).Methods:(1)The GA patients were divided into 4 groups:44 acute gout controls(AG),41 inter-critical gout patients(IG),30 chronic gout patients(CG)and 48 healthy patients,collecting their peripheral blood and laboratory indexes.(2)Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression levels of PFKM,PFKL and PFKP mRNA in PBMCs of 4 groups.(3)Enzyme-linked immunosorbent assay(ELISA)and Western blot was used to measure PFKM,PFKL and PFKP proteins in four groups of PBMCs.(4)SPSS 22.0 statistical software was used for analysis.The statistical description of normal measurement data was measured by mean±standard deviation(x ±s),and the statistical description of non-normal measurement data was measured by median(quartile spacing)[M(Q)].The measurement data were compared by rank sum test and one-way ANOVA.The least significant difference method(LSD method)was used to compare the two groups.The correlation analysis between variables was performed by Spearman correlation analysis.A P value less than 0.05 was considered to denote statistical significance.Results:(1)The ESR of the AG group was significantly higher than that of the IG and CG groups(p<0.05),and the ESR of the CG group was apparently higher than IG group(p<0.05).The concentrations of hsCRP and GLOB in the AG group were apparently higher than those in the HC,IG and CG groups(p<0.05),and the concentration of GLOB in the CG group was more than IG group(p<0.05).The number of WBC and GR cells in AG and CG groups were more remarkable than those in HC and IG groups(p<0.05).The EOs in the AG group was lower than those in the HC and IG groups(p<0.05),and similar comparison was found in the CG group and HC group(p<0.05).MCH concentrations in the AG,IG,and CG groups were significantly lower than those in the HC group(p<0.05).The concentrations of MCHC in the AG and CG groups were lower than those in the HC and IG groups(p<0.05).The plasma concentrations of AST,ALT and GGT in AG and IG groups were apparently higher than HC group(p<0.05),and the plasma concentration of GGT in CG group was also significantly higher than that in HC group(p<0.05).The plasma concentration of HDL-C in IG group was significantly lower than that in AG and HC groups(p<0.05).Conversely,the plasma concentration of VLDL-C in AG and IG groups were remarkable higher than that in HC group(p<0.05).The concentration of Crea in the AG and CG groups were significantly higher than that in the HC group(p<0.05).Similarly,the CG group was significantly higher than the IG group(p<0.05).The UA concentration in the AG,IG,and CG groups were significantly higher than that in the HC group(p<0.05).The GLU concentration in the AG group was significantly higher than that in the HC group(p<0.05).(2)The expression levels of PFKM and PFKP mRNA in PBMCs of GA patients were not significantly different from those in healthy controls(P>0.05).In addition,the expression levels of PFKL mRNA in PBMCs of patients with GA was lower than that of healthy controls(P>0.05).Further comparison among four groups,the expression of PFKL mRNA in AG group was significantly lower than that in HC group and CG group(P<0.05,P<0.01,respectively),but there was no statistical difference was found between AG and IG groups(p>0.05).The expression level of PFKL in IG group was significantly lower than that in CG group(P<0.05),but there was no significant difference between IG group,HC group and AG group(P>0.05).The PFKL mRNA expression in the CG group was significantly higher than that in the AG and IG groups(P<0.05),however,similar improvements in CG group and healthy controls subsets of both groups were not observed.(3)The expression of PFKM protein in PBMCs of patients with GA was lower than that of healthy controls(P<0.05).Compared with the two groups,the expression levels of PFKM protein in AG group and IG group were significantly lower than that in HC group(both P<0.05),but AG group and There was no significant difference in IG group(P>0.05).The expression of PFKM protein in CG group was lower than that in HC group,higher than AG group and IG group,but the difference was not statistically significant(P>0.05).There was no significant difference in the expression of PFKL and PFKP protein in PBMCs of patients with GA(P>0.05).(4)Spearman correlation analysis between PFKL mRNA and laboratory-related indicators showed that the expression of PFKL mRNA in the AG group was negatively correlated with PLT(r=-0.341,P=0.045).The expression of PFKL mRNA in the CG group was positively correlated with hsCRP(r=0.648,P=0.043).Conclusion:(1)Different stages of gout have different inflammatory reactions.Gout affects the functions of liver and kidney,and affects the metabolism of red blood cell hemoglobin and glucose at the same time.It is proved that the importance of maintaining uric acid in the intermittent and chronic stages of gout is up to standard.(2)It was confirmed that the genes and proteins of three subtypes of PFK1 are expressed in mononuclear cells of patients with primary gouty arthritis and healthy people,and no obvious defects were found in patients with primary gout.This may be related to type VII of glycogen storage disease.And this is a really disctinction in secondary gout caused by glycogen storage disease.(3)In the process of gout disease,the PFKM,PFKL gene transcription level and protein expression are inconsistent.Does platelet and C-reactive protein have different regulation with PFKL mRNA expression during gout disease? Further research is needed to confirm. |
