| Objective This article studies the expression of apoptosis related protein c-FLIP in peripheral blood mononuclear cells of patients with rheumatoid arthritis at different period,in order to provide the idea that treating and looking for new drug effected targets.Methods 1.Peripheral blood samples from patients and healthy control were collected.ESR was detected by fully automatic blood sedimentation instrument,RF and hs-CRP were detected by full automatic immune turbidimetric instrument and anti-CCP antibody was detected by ELISA.2.m RNA expression of c-FLIP,NF-?B,Bcl-2 and FADD,TRAIL,caspase-8 were detected by Real-time PCR from peripheral blood mononuclear cells.3.c-FLIP,caspase-8,FADD and TRAIL were checked by western blot from peripheral blood mononuclear cells.Results 1.The results of ESR,hs-CRP,RF,anti CCP in peripheral blood at different groups with RA: 1.1 ESR value in Low activity(L)group,middle activity(M)group and high activity(H)group were 9.67 ± 6.22 mm/h,57.38 ± 26.62 mm/h and 50.80 ± 26.80 mm/h respectively which was significantly higher than which in healthy controls(3.90± 1.21 mm/h);(P=0.023,0.001,0.002),The group M and H were obviously higher than that of the group L(P=0.009,0.010),however,there was no significant difference between group M and H(P> 0.05).1.2 Serum hs-CRP value in Low activity(L)group,middle activity(M)group and high activity(H)group were 1.12 ± 1.21 IU/ml,26.73 ± 16.30 IU/ml and 31.16 ± 9.77 IU/ml respectively which was higher than that of healthy controls(0.63±0.23 IU/ml)(P=0.039,0.001,0.002);The group M and H was obviously higher than that of the group L(P=0.003?0.002).1.3 Serum RF in group L,M and H were 383.72 ± 167.80 mg/L,142.33 ± 126.02 mg/L,233.75 ± 170.96 mg/L which was higher than healthy controls 2.81 ± 1.76 mg/L(P<0.05,P<0.01,P<0.01 respectively).1.4 Serum anti CCP values in group L,M and H were 963.13 ± 617.85 RU/ml,468.79 ± 329.39 RU/ml and 202.45 ± 119.33 RU/ml,which were higher than that of healthy controls(5.13 ± 2.47 RU/ml)(P values were 0.001,0.013,0.015).1.5 The sensitivity for RA diagnosis was 87% by means of RF detection and its specificity was 79%,however,the sensitivity for RA diagnosis was 68% and specificity was 100% with the combination detection of ESR,hs-CRP,anti CCP and RF.2.The m RNA expression level of c-FLIP,NF-?B,BCL-2 and FADD,TRAIL,caspase-8 from PBMC at different groups of RA: 2.1 The relative level of c-FLIP m RNA in low activity group(L),middle activity group(M)and high activity group(H)were 1.27 ± 0.28,1.32 ± 0.34 and 1.93 ± 0.40 respectively which was higher than that of healthy controls(1.08 ± 0.12)(P= 0.035,0.034,0.030).2.2 FADD m RNA in group L,group M,group H were 0.70 ± 0.16,1.02 ± 0.24 and 1.11 ± 0.18 which was higher than healthy controls(0.44 ± 0.06)(P= 0.037,0.028,0.023).2.3 TRAIL m RNA in group M was 3.26 ± 0.78 which was much higher than healthy controls(1.30 ± 0.20)(P= 0.028,and those of group L and group H(1.56± 0.37 and 1.83 ± 0.26 respectively)was slightly higher than controls(P<0.05).2.4 NF-?B m RNA in group L and M were 0.98 ± 0.19 and 0.76 ± 0.26 which was higher than healthy controls(0.56 ± 0.08)(P= 0.018,0.029),however,that of in group H(0.15 ± 0.09)was less than control.2.5 Caspase-8 m RNA in group L,group M,group H were 2.77 ± 0.97,4.52 ± 0.85,and 2.13 ± 0.44 which was higher than healthy controls(1.04 ± 0.13)(P= 0.023,0.012,0.032)2.6 Bcl-2 m RNA in group L(1.02 ± 0.35)increased compared with control(0.32 ± 0.18)(P= 0.009),and decreased in group M and H(0.08 ± 0.01,0.05± 0.02).2.7 There was a positive correlation between C-FLIP and FADD,negative correlation between C-FLIP and TRAIL,NF-?B,caspase-8,Bcl-2 by means of bivariate regression analysis.3.The Protein level of c-FLIP?caspase-8?FADD?TRAIL in PBMC from different groups of RA patients: 3.1 C– FLIP expression in the group M was significantly increased than control(P=0.001)with no difference in group L and H(P=0.062,0.053).3.2 Caspase-8 expression in the group M and H was significantly enhanced than control(p=0.003,0.001)with no difference in group L(p> 0.05).3.3 The expression of FADD in the group M and H was significantly increased than control(P=0.012,0.037)with no difference in group L(p> 0.05).3.4 The expression of TRAIL in M group was significantly increased than group L,H and control(P<0.05)with no difference in group L.Conclusions 1.Combination detection of ESR,hs-CRP,RF,anti CCP increased the diagnostic sensitivity and specificity of RA,and showed the advantages in valuation of disease development.2.In peripheral blood mononuclear cells of patients with RA,m RNA of apoptosis associated protein c-FLIP is positively correlated with the progression of disease,and can be provided reference for RA appraisal.3.During the different active stages of RA,in peripheral blood mononuclear cells,the expression of FADD,Caspase-8,TRAIL,and NF-?B,Bcl-2 are higher and higher.But the role of apoptosis in RA's mechanism is requested to keep studying. |
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