The Expression And Clinical Significance Of Superoxide Dismutase In The Patients With Primary Gouty Arthritis

OBJECTIVE:To measure the expressions of SOD1 and SOD2 genes,proteins,and protease activities in peripheral blood mononuclear cells(PBMCs),including the protein concentrations and protease activities of SOD3 in plasma,in the patients with Gout at different stages.The differences of SOD expressions and the possible roles of SOD in the patients with primary gouty arthritis were investigated preliminarily.Methods:(1)Collected the clinical and laboratory data in 48 cases of acute gouty arthritis(AG),48 cases of intermittent gouty arthritis(IG),48 cases of chronic gouty arthritis(CG)and 48 cases of healthy controls(HC).(2)To measure the expressions of SOD1 and SOD2 mRNA in PBMCs by real-time quantitative polymerase chain reaction(RT-qPCR).(3)Western blot(WB)was used to measure the expressions of SOD1 and SOD2 proteins in PBMCs in 14 cases of AG,14 cases of IG,14 cases of CG and 14 cases of HC.(4)The enzyme-linked immunosorbent assay(ELISA)was used to measure the SOD3 protein concentrations in the plasma in 14 cases of AG,14 cases of IG,14 cases of CG and 14 cases of HC.(5)The enzyme activity of SOD1,SOD2 in PBMC and SOD3 in plasma was measured by WST in 14 cases of AG,14 cases of IG,14 cases of CG and 14 cases of HC.Statistical analysis was performed using SPSS 19.0 software.P<0.05 was considered the difference to be statistically significant.Results:(1)The levels of SOD1 mRNA in PBMCs were significantly differences between the group of AG,CG,IG and HC(H=62.554,P<0.001).Performing a pairwise comparison,the expression levels of SOD1 mRNA were significantly higher in the gout patients(AG,CG,and IG)than that in the HC group(P<0.001);The levels in the AG group were significantly lower than that in the CG and IG groups(P<0.001);there was no statistical difference between the CG and the IG groups(P>0.05).The SOD2 mRNA levels in PBMCs were significantly different in the AG,CG,IG and HC groups(H=39.823,P<0.001).Performing a pairwise comparison,SOD2 mRNA levels were significantly higher in the gout patients(AG,CG,IG)than that in the HC group(P<0.001);The levels in the CG group was significantly lower than that in the AG and IG groups(P<0.05);there was no statistical difference between the AG and the IG groups(P>0.05).(2)The SOD1 protein levels in PBMCs was significantly different in the AG,CG,IG and HC groups(H=7.742,P<0.05).Performing a pairwise comparison,the levels of SOD1 protein were lower in the CG group than that in the AG and IG groups(P<0.05);there was no statistical difference in the HC group between the AG,CG and IG groups(P>0.05);there was no statistical difference between the AG and the IG groups(P>0.05).The SOD2 protein levels in PBMCs was significantly different between AG,CG,IG,and HC groups(H=11.167,P<0.01).Performing a pairwise comparison,the levels of SOD2 protein was significantly lower in the CG group than that in the AG,IG and HC groups(P<0.05);there was no statistical difference between the HC group and the AG,IG groups(P>0.05);there was no statistical difference between the AG and the IG group(P>0.05).(3)For SOD 1 enzyme activity in PBMCs,there was no statistical difference between AG,CG,IG and HC groups(F=0.462,P>0.05).For SOD2 enzyme activity in PBMCs,there was no statistical difference between AG,CG,IG and HC groups(F=1.560,P>0.05).(4)The concentration of SOD3 protein in plasma was significantly different in the AG,CG,IG and HC groups(F=2.892,P<0.05).The plasma SOD3 protein in AG group was significantly higher than that in HC group(P<0.01),but there was no statistical difference between the other groups(P>0.05).(5)There was statistical difference in the SOD3 enzyme activity(U/ng)in the AG,CG,IG and HC groups(F=8.541,P<0.05).Performing a pairwise comparison,the SOD3 enzyme activity was significantly lower in the AG group than that in the HC group(P<0.01);but there was no statistical difference between the other groups(P>0.05).(6)The SOD2 mRNA levels in the AG group was correlated negatively with LY(r=-0.576,P=0.031);the SOD1 protein was correlated positively with WBC(r=0.568,P=0.034)and NE(r=0.559,P=0.038),LDLC(r=0.618,P=0.043),APOB(r=0.688,P=0.019);the SOD2 protein was correlated positively with LDLC(r=0.709,P=0.015);the SOD2 protease activity was correlated negatively with LDLC(r=-0.636,P=0.035);the SOD3 protein concentration was correlated positively with WBC(r=0.574,P=0.032),NE(r=0.570,P=0.033),MO(r=0.637,P=0.014);and the SOD3 protease activity was correlated negatively with WBC(r=-0.563,P=0.036),NE(r=-0.563,P=0.036),and MO(r=-0.717,P=0.004).(7)The SOD2 mRNA levels in PBMCs in the IG group was correlated positively with TG(r=0.665,P=0.026),LDLC(r=0.618,P=0.043)and APOB(r=0.636,P=0.035);the SOD1(r=-0.653,P=0.011)and SOD2(r=-0.600,P=0.023)proteins was correlated negatively with ESR.(8)The SOD2 mRNA levels in the CG group was correlated negatively with WBC(r=-0.666,P=0.009),MO(r=0.666,P=0.009),and LY(r=-0.570,P=0.033).the SOD2 mRNA levels was correlated positively with TG(r=0.900,P=0.001)and APOB(r=0.636,P=0.035);the SOD3 protein concentration was correlated negatively with ALT(r=-0.565,P=0.035).(9)The SOD2 mRNA levels in PBMCs in the HC group was correlated negatively with WBC(r=-0.539,P=0.047),and was correlated positively with TG(r=0.567,P=0.043).The SOD1 protein was correlated positively with ALT(r=0.777,P=0.002)and AST(r=0.562,P=0.046),and was correlated negatively with LDLC(r=-0.585,P=0.046).The SOD2 protein was correlated positively with ALT(r=0.672,P=0.012),AST(r=0.667,P=0.013),LDLC(r=-0.669,P=0.017).The SOD1 protease activity was correlated negatively with CRP(r=-0.714,P=0.047);the SOD2 protease activity was correlated negatively with ALT(r=-0.592,P=0.033),and was correlated positively with LDLC(r=0.676,P=0.016).The SOD3 protein was correlated negatively with LY(r=-0.538,P=0.047).Conclusion:(1)The PBMCs of primary gout patients are in an oxidative stress state.(2)The primary gout patients are in an oxidative stress state.(3)SOD is involved in the acute inflammatory response of gouty arthritis.(4)The fatty disorders,vascular endothelial damages,and atherosclerosis in patients with primary gout may be related to oxidative stress in the mitochondria.