| Objective:The absolute number of Th17,Treg cells,Th17/Treg ratio,serum MMP-3,TIMP-1and MMP-3/TIMP-1 ratios in peripheral blood of rheumatoid arthritis were measured to investigate its role in the pathogenesis and mechanism.Methods:Eighty-seven RA patients diagnosed by the Department of Rheumatology and Immunology,Second Hospital of Shanxi Medical University from April 2017 to January2018 were recruited as the RA group.30 patients from the physical examination center of our hospital were recruited as the control group.The gender and age of the two groups were matched.According to the score of DAS28,RA patients were divided into active group(67 cases)and remission stage group(20 cases).Active group was divided into 3groups.Among them,25 cases were highly active,20 cases were moderate active,22 cases were low active,and the gender and age of the four groups were matched.Serum samples of all subjects were collected.The absolute number of Th17 and Treg cells was measured by flow cytometric analysis.Serum MMP-3,TIMP-1,and anti-CCP antibodies were measured by ELISA.RF and CRP were measured by immunoturbidimetry.ESR was measured by Wei's method.All data were entered into the EXCEL table,SPSS23.0statistical software was used for data processing and statistical analysis.The Kolmogorov-Smirnov test was used to test the normality.The normal distribution data was expressed as the mean±variance(x±s).One-sample ANOVA and LSD-t test wereused to compare the mean of the samples,and Pearson correlation analysis was used to test the correlation.The test level was ? = 0.05,P <0.05 was considered statistically significant.Results:1.Comparisons of Th17,Treg,Th17/Treg,MMP-3,TIMP-1,MMP-3/TIMP-1between RA and healthy control groups.The levels of Th17,Treg,Th17/Treg,MMP-3,TIMP-1 and MMP-3/TIMP-1 in RA group were(6.70±5.44)/ul,(27.01±15.08)/ul,(0.31±0.26),(3.01±0.40)ng/ml,(2.81±0.31)ng/ml and(1.08±0.16);in control group were(4.44±4.11)/ul,(33.07±11.86)/ul,(0.13±0.10),(2.65±0.34)ng/ml,(2.65±0.30)ng/ml and(1.01±0.14).The levels of Th17,Th17/Treg,MMP-3,TIMP-1,MMP-3/TIMP-1 in RA group were higher than those in control group(P<0.05 or P<0.001).The Treg in RA group was lower than that in control group.There was a statistically significant difference between the two groups(P<0.05).2.To compare the serum lymphocyte related parameters: the number of Th17 cells,Treg cells and their ratios in RA patients with low,middle and high active period.There was no significant difference in the number of Th17 cells among the three groups(F=0.797,P>0.05);the number of Treg cells was statistically significant(F=3.342,P<0.05),and the high activity group was lower than the remission period.There was no significant difference between the middle active group and the remission group and low activity group(P<0.05);the ratio of the number of Th17 and Treg cells in each group was statistically significant(F=2.902,P<0.05).The high activity group was higher than the remission group(P<0.05)and the low activity group(P<0.05),but there was no significant difference between the middle activity group and the remission group and low activity group.(P>0.05).3.Comparison of three serum markers(MMP-3,TIMP-1 and MMP-3/TIMP-1)in patients with RA in the low,middle and high activity groups: The serum levels ofMMP-3 in RA groups were significantly different(F=3.851,P<0.05).The high activity group was higher than the low,middle activity group and remission group(P<0.05).There was no significant difference in the mid-activity group and the remission group and the low activity group.(P>0.05);Serum TIMP-1 concentrations in each group were not statistically significant(F=1.446,P>0.05);MMP-3/TIMP-1 in each group had statistically significant(F=2.801,P< 0.05).The high activity group was higher than the remission group and low activity group(P<0.05),while the middle activity group was not significantly different from the remission group and low activity group(P>0.05).4.Correlativity between peripheral blood lymphocyte markers and clinical parameters in RA group: There was no correlation between Th17 cell number and ESR,CRP,RF,anti-CCP,HAQ;Treg cell number was negatively correlated with ESR,CRP,HAQ(r=-0.352,P=0.001;r=-0.248,P=0.021;r=-0.216,P=0.044),no correlation with RF and anti-CCP;Th17/Treg was positively correlated with ESR,CRP,and HAQ(r=0.277,P=0.009;r=0.227,P=0.034;r=0.258,P=0.016),and has no correlation with RF and anti-CCP.5.Correlations between serum MMP-3 and TIMP-1 levels,MMP-3/TIMP-1 and clinical parameters in RA group were positively correlated with ESR,CRP,anti-CCP and HAQ(r=0.274,P=0.010;r=0.267,P=0.012;r=0.220,P=0.041;r=0.213,P=0.048),no correlation with RF;TIMP-1 had no correlation with ESR,CRP,RF,anti-CCP,HAQ;There was a positive correlation between MMP-3/TIMP-1 and ESR,CRP,anti-CCP and HAQ(r=0.249,P=0.020;r=0.248,P=0.021;r=0.292,P=0.006;r=0.222,P=0.039).There is no correlation with RF.6.The correlation analysis of Th17,Treg,Th17/Treg,MMP-3,TIMP-1 and MMP-3/TIMP-1 in peripheral blood of RA patients showed that Th17 cell number in peripheral blood of RA patients was positively correlated with Th17/Treg(r=0.681,P<0.001),Treg cells were negatively correlated with Th17/Treg(r=-0.279,P=0.009),Treg cells were negatively correlated with MMP-3 concentration(r=-0.222,P=0.039),there was a positive correlation between the concentration of MMP-3 and TIMP-1(r=0.213,P=0.048).There was a positive correlation between MMP-3 andMMP-3/TIMP-1(r=0.711,P<0.001).TIMP-1 and MMP-3 /TIMP-1 was negatively correlated(r=-0.521,P<0.001).There was no correlation between MMP-3 and Th17/Treg,Treg and MMP-3/TIMP-1,Th17/Treg and MMP-3/TIMP-1.Conclusion:There is a negative correlation between the number of Treg cells and the pathogenesis and progression of RA.The balance of Th17 to Treg ratio plays an important role in the course of the disease and has a positive correlation with the disease activity.There is a positive correlation between the concentration of MMP-3 and the pathogenesis and progression of RA;the balance of MMP-3/TIMP-1 plays an important role in it and is positively correlated with the disease activity.The decrease of Treg cells and the increase of MMP-3 concentration may trigger the occurrence and aggravation of RA,and there is a certain correlation between them,and they influence each other to strengthen the autoimmune response of RA. |
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