| Background:Rheumatoid arthritis(RA) is a chronic and systemic autoimmune disorder. Multi arthritis as the major clinical manifestation in RA leads to joints and other tissue or organ damage. RA is one of the main causes of human lossing of labor and disability. At present, the pathogenesis of RA is unclear. It was considered that environmental factors, genetic susceptibility and immune dysfunction were revolved. micro RNAs(mi RNAs) is a class of short noncoding RNA that is encoded by genomes of the higher eukaryotes. mi RNAs play a negative role to gene expression in the post transcription,which pair target m RNA by complementary base to degradate of the target m RNA or prevent protein translation.. A large number of studies showed that mi RNAs had played an important role in cell development, hematopoiesis, organ formation, cell proliferation, apoptosis and tumorigenesis. The abnormal expression of mi RNA in tumor, inflammation and autoimmune diseases has become a research hotspot in recent years.Objectives:To detect the expression levels of mi R-146 a and mi R-23 b in peripheral blood mononuclear cells(PBMCs) in patients with RA and healthy women control, and then to analyse the difference expression level of mi R-146 a and mi R-23 b between RA group and healthy control group. To explore the effect of mi R-146 a and mi R-23 b on RA pathogenesis by determining the relationship between the expression level of mi R-146 a or mi R-23 b and RA clinical indicators, such as interleukin-17(IL-17), erythrocyte sedimentation rate(ESR), C-reactive protein(CRP), anti cyclic citrullinated peptide(CCP) antibody, rheumatoid factor(RF) of plasma and disease activity score(DAS) of RA patients. Furthermore, to evaluate the effect of estrogen on expression of mi R-146 a, mi R-23 b and IL-17, and to reveal the mechanism that estrogen affects on RA.Methods:1. 34 patients with RA in active stage were investigated as RA group and the fasting peripheral venous blood was collected. These blood samples were treated by heparin anticoagulant and plasma was isolated and conserved at-80℃. At the same time, PBMCs were separated. Clinical characteristics of the patients with RA were collected. 40 healthy women were collected as the normal control group.2. Total RNA was extracted and the basal expression level of mi R-146 a and mi R-23 b was measured by real-time quantitative polymerase chain reaction(RT-q PCR). The results were compared between RA group and normal control group.3. The concentration of IL-17 in plasma was measured by enzyme-linked immunosorbent assay(ELISA). The difference was compared between RA group and normal control group. The correlation between mi R-146 a or mi R-23 b with IL-17, ESR, CRP, anti-CCP antibody, RF, or DAS of patients with RA was evaluated.4. Another part of PBMCs were treated with or without(blank control) estradiol(E2) for 6h, the culture supernatant and cells were collected respetively. The culture supernatant was conserved at-80℃. Total RNA was extracted and the expression level of mi R-146 a and mi R-23 b was measured by RT-q PCR. The results were compared between treated group and untreated group.5. The concentration of IL-17 in culture supernatant was measured by ELISA, the difference was compared between treated group and untreated group. The correlation between the expression level of mi R-146 a or mi R-23 b with IL-17 in culture supernatant was evaluated.Results:1. It was indicated that the basal expression level of mi R-146 a in PBMCs [5.12E-03(2.16E-03, 14.41E-03) vs 3.48E-03(2.24E-03, 5.15E-03), P=0.040], mi R-23 b in PBMCs [0.24E-03(0.13E-03,0.67E-03) vs 0.16E-03(0.08E-03,0.31E-03), P=0.041] and the concentrations of IL-17 in plasma [1.80 pg/ml(0.84, 4.89 pg/ml) vs 0.85 pg/ml(0.24, 3.51 pg/ml), P=0.047] in patients with RA were increased significantly than the healthy women.2. A positive correlation was found between the expression level of mi R-146 a in PBMCs from female patients with RA with ESR(R=0.377, P=0.030), but no significant correlation was found between mi R-146 a with IL-17 in plasma(R=-0.184, P=0.332), CRP(R=0.067, P=0.717), anti CCP antibody(R=-0.116, P=0.520), RF(R=0.000, P=0.999), DAS(R=0.289, P=0.097).There was no significant correlation between mi R-23 b with IL-17(R=0.000, P=0.999), CRP(R=-0.065, P=0.722), ESR(R=0.041, P=0.820), anti CCP antibody(R=0.022, P=0.902), RF(R=-0.005,P=0.979), DAS(R=-0.149, P=0.401).3. PBMCs were treated with or without E2 at the concentration of 1×10-7mol/L for 6 hours, then the results were compared:(1) Patients with RA: No significant difference was found in the expression level of mi R-146 a [4.88E-03(2.88E-03, 14.13E-03) vs 4.78E-03(1.82E-03, 13.74E-03), P=0.360], mi R-23 b [0.24E-03(0.08E-03, 0.78E-03) vs 0.24E-03(0.08E-03, 0.66E-03), P=0.755] and the concentrations of IL-17 [0.86 pg/ml(0.72, 1.01 pg/ml) vs 0.75 pg/ml(0.65, 0.87 pg/ml), P=0.078]. It was indicated that estrogen had no significant effect on the expression of mi R-146 a, mi R-23 b or IL-17 in PBMCs in patients with RA.(2) The normal control: No significant difference was found in the expression level of mi R-146 a [2.75E-03(2.01E-03, 4.91E-03) vs 3.00E-03(1.99E-03, 5.20E-03), P=0.563], mi R-23 b [0.12E-03(0.06E-03, 0.23E-03) vs 0.13E-03(0.05E-03, 0.30E-03), P=0.485] and the concentrations of IL-17[0.76 pg/ml(0.65, 0.86 pg/ml) vs 0.76 pg/ml(0.72, 0.86 pg/ml), P=0.355]. It was indicated that estrogen had no significant effect on the expression of mi R-146 a, mi R-23 b or IL-17 in PBMCs from healthy population.(3) RA group: No significant correlation was found between the expression level of mi R-146 a with IL-17 in culture supernatant(R=-0.257, P=0.355), mi R-23 b with IL-17(R=-0.414, P=0.125) from treated PBMCs in patients with RA.(4) The normal control group: No significant correlation was found between the expression level of mi R-146 a with IL-17 in culture supernatant(R=0.433, P=0.064), mi R-23 b with IL-17(R=-0.233, P=0.338) from treated PBMCs in healthy females.Conclusions:1. The basal expression level of mi R-146 a, mi R-23 b in PBMCs and the concentration of IL-17 in plasma in patients with active RA were significantly higher than that of healthy women. It was indicated that the increased expression of mi R-146 a, mi R-23 b as well as IL-17 may be relative to the pathogenisis of RA.2. A positive correlation was found between the expression level of mi R-146 a in PBMCs with ESR from female patients with RA, it was indicated that mi R-146 a in PBMCs probably associated with disease activity of RA.3. No significant influence of E2 at 1×10-7mol/L on the expression of mi R-146 a, mi R-23 b or IL-17 in PBMCs at 6 hours in patients with RA or healthy women was found. Besides, there was no obviously synergistic or antagonistic effect had found between mi R-146 a or mi R-23 b with IL-17. |
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