| Objective To detect the expression of miR326 and Ets-1 in CD4+T cells cells from patients with SLE, to analyze the relationship between the Th17 ratio and the expression of miR326 and Ets-1 mRNA, to explore the potential pathogenesis of miR326 in SLE.Methods According to 2009 American College of Rheumatology (ACR) SLE diagnosis standards,40 SLE patients (containing 5 male and 35 females, mean age 36.00±12.58) were screened from the outpatient and inpatient of the Anhui Provincial Hospital Rheumatology Department. The lab index and the clinical data had been recorded carefully. Based on systemic lupus erythematosus disease activity index (SLEDAI) patients were divided into the active group and the remitting group, the active group with SLEDAI>4 and the remitting group with SLEDAI≤4, and the active group were divided into mild active group (4<SLEDAI≤9), moderate active group (9<SLEDAI≤14) and severe active group (SLEDAI>14). Fourteen healthy volunteers without autoimmune disease were chosen as controls, sex and age matched. Flow cytometry was used to inspect the ratio of Th17 cells (CD4+IL-17A+)/CD4+T cells. Magnetic activated cell sorting was used to enrich the CD4+T cells, and reverse Transcription Polymerase Chain Reaction (RT-PCR) was used to inspecte the relative expression of miR326 and Ets-1 mRNA in CD4+T cells. For the quantitative data, t test and rank sum test were used to analysis the difference between the two groups, comparison between the three groups were analyzed by one-way ANOVA, q test and Kruskal- Willis test. Pearson test was used to analysis the data wich in accordance with normal distribution. Spearman test was used to analysis the correlation between miR326. Ets-1 mRNA, Th17 proportion and clinical data.Results 1. Compared to the healthy controls, the miR326 expression level in the CD4+T cells of SLE patients was increased obviously (.P=0.003), and the expression level was higher in the active SLE group than in remitting SLE group(P=0.000).2. The Ets-1 mRNA expression level in CD4+T cells of SLE patients was reduced obviously than the controls (^=0.002), and the expression in the active SLE group was lower than in remitting SLE group (P=0.001).3. Compared to the controls, the ratio of Thl7 cells/CD4+T cells in SLE patients was increased obviously (P=0.004), and significant difference was found between the active group and the remitting group. 4. The expression of miR326 had a negative correlation with the Ets-1 expression (P<0.001) and a positive correlation with the ratio of Th17 cells (P=0.001). There was also a negative correlation between Ets-1 expression and the ratio of Th17 cells (P=0.003).5. The miR326 expression and the ratio of Th17 cells both had a positive correlation with the SLEDAI score, while the Ets-1 expression had a negative correlation with the SLEDAI score.6. In SLE patients who suffer from erythra, hematologic system or kidney impairment, the miR326 expression and the ratio of Th17 cells increased significantly, while the Ets-1 expression induced compared to the rest SLE patients.7. The miR326 expression and the ratio of Th17 cells positively regulated with anti-dsDNA antibodies level and anti-nucleosome antibodies level, and negatively regulated with C3 and C4. On the contrary the Ets-1 expression had a negative regulation with anti-dsDNA antibodies and anti-nucleosome antibodies and a positive regulation with C3 and C4.Conclusion The miR326 expression of CD4+T cells increases obviously in SLE patients and positively correlates to the ratio of Th17 cells and the disease activity index, but has a negative correlation with Ets-1 expression. The results prompt that miR326 could participate in the disease activity of SLE through inhibiting the Ets-1 gene expression and promoting the differentiation of Th17 cells. |
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