| 李家江,王守鹏,张兆川,等.微小RNA-146a 靶向调控Krüppel 样转录因子7对强直性脊柱炎T 细胞炎症反应及自噬的影响[J].安徽医药,2022,26(8):1611-1614. |
| 微小RNA-146a 靶向调控Krüppel 样转录因子7对强直性脊柱炎T 细胞炎症反应及自噬的影响 |
| Effect of miR-146a on T cell inflammatory response and autophagy in patients with ankylos?ing spondylitis by targeting KLF7 expression |
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| DOI：10.3969/j.issn.1009-6469.2022.08.029 |
| 中文关键词: 脊柱炎，强直性 微小RNA-146a Krüppel样转录因子7 T细胞 炎症 自噬 |
| 英文关键词: Spondylitis,ankylosing MiRNA-146a Krüppel-like factor T cells Inflammation Autophagy |
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| 中文摘要: |
| 目的探讨微小RNA-146a(miR-146a)是否通过靶向调控Krüppel样转录因子7(KLF7)的表达而影响强直性脊柱炎病人T细胞炎症反应及自噬。方法采用实时荧光定量聚合酶链反应(qRT-PCR)检测强直性脊柱炎病人T细胞与健康人T细胞中miR-146a的表达量；分别将anti-miR-NC、anti-miR-146a、pcDNA3.1、pcDNA3.1-KLF7转染至强直性脊柱炎病人T细胞；采用酶联免疫吸附法(ELISA)检测白细胞介素-6(IL-6)、白细胞介素-17(IL-17)、白细胞介素-23(IL-23)的水平；双荧光素酶报告实验验证miR-146a与KLF7的靶向结合关系；蛋白质印迹法(Western blotting)检测KLF7、自噬相关基因5(ATG5)、Beclin-1的表达量。结果与健康人T细胞比较，强直性脊柱炎病人T细胞中miR-146a的表达量升高［(1.00±0.08)比(2.74±0.13)，t=53.54，P<0.05］；与anti-miR-NC组比较，anti-miR-146a组IL-6［(823.17±14.10)ng/L比(372.31±11.03)ng/L］、IL-17［(901.00±16.29)ng/L比(428.39±12.34)ng/L、IL-23［(886.38±15.11)ng/L 比(401.61±11.75)ng/L］的水平降低(t=43.62，40.06，43.87，P<0.05)，ATG5［(0.33±0.03)比(0.79±0.07)］、Beclin-1［(0.42±0.04)比(0.91±0.08)］蛋白水平升高(t=10.46，9.49，P<0.05)；双荧光素酶报告实验证实miR-146a可靶向结合KLF7；转染pcDNA3.1-KLF7可明显降低IL-6［(823.17±14.10)ng/L比(477.37±12.01)ng/L］、IL-17［(901.00±16.29)ng/L比(558.67±13.16)ng/L］、IL-23［(886.38±15.11)ng/L比(531.69±12.75)ng/L］的水平(t=32.34，28.31，31.07，P<0.05)，提高ATG5［(0.32±0.03)比(0.60±0.06)］、Beclin-1［(0.44±0.04)比(0.78±0.07)］的表达水平(t=7.23，7.30，P=0.002)。结论抑制miR-146a表达可通过靶向KLF7而抑制强直性脊柱炎病人T细胞炎症反应及促进细胞自噬。 |
| 英文摘要: |
| Objective To investigate whether miR-146a affects T cell inflammation and autophagy in patients with ankylosing spon?dylitis by regulating the expression of KLF7.Methods qRT-PCR was used to detect the expression of miR-146a in T cells from pa?tients with ankylosing spondylitis and healthy human T cells. anti-miR-NC, anti-miR-146a, pcDNA3.1, and pcDNA3.1-KLF7 were transfected into T cells of patients with ankylosing spondylitis, respectively. ELISA was used to detect the levels of IL-6, IL-17 and IL-23. The dual luciferase reporting experiment verified the targeted binding relationship between miR-146a and KLF7. The expression levels of KLF7, ATG5 and Beclin-1 were detected by Western blotting.Results Compared with healthy human T cells, the expression of miR-146a in T cells of patients with ankylosing spondylitis increased significantly [(1.00±0.08 ) vs. (2.74±0.13) ,t=53.54, P <0.05].Compared with the anti-miR-NC group, the levels of IL-6 [(823.17±14.10)ng/L vs. (372.31±11.03)ng/L], IL-17 [(901.00±16.29)ng/L vs.(428.39±12.34)ng/L], and IL-23 [(886.38±15.11)ng/L vs. (401.61±11.75)ng/L] in the anti-miR-146a group were significantly reduced (t=43.62, 40.06, 43.87, P<0.05), and the levels of ATG5 [(0.33±0.03) vs. (0.79±0.07) ]and Beclin-1 [(0.42±0.04) vs. (0.91±0.08)] proteins were significantly increased (t=10.46, 9.49, P<0.05). Double luciferase reporting experiments confirmed that miR-146a could target KLF7. Transfection of pcDNA3.1-KLF7 could significantly reduce the levels of IL-6 [(823.17±14.10) vs. (477.37±12.01)ng/L, IL-17[(901.00±16.29)ng/L vs. (558.67±13.16)ng/L, IL-23 [(886.38±15.11) vs. (531.69±12.75)ng/L,t=32.34, 28.31, 31.07, P<0.05], and in?crease the expression levels of ATG5 [(0.32±0.03) vs. (0.60±0.06)] and Beclin-1[(0.44±0.04) vs. (0.78±0.07),t=7.23, 7.30, P=0.002].Conclusion Inhibition of miR-146a expression can inhibit T cell inflammatory response in patients with ankylosing spondylitis and promote cell autophagy by targeting KLF7. |
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