miR-17-5p及其靶基因ANKH在强直性脊柱炎发病过程中作用机制的研究

The pathogenesis of ankylosing spondylitis is unknown. Loss of function ANKH, will appear similar to the changes in ankylosing spondylitis. There is no evidence that ANKH gene mutation occur in patients with AS. Others results show，the expression levels of miR-17-5p were much higher in AS group than those in control group. We find that one of target genes of miR-17-5p is ANKH by test (Dual Luciferase Reporter Gene Assay ) and analysis in Targetscan and miRanda. Further, we observed that expression of ANKH in Fibroblast cells was associated with the overexpression of miR-17-5p. Now, we have established our hypothesis: Regulation of ANKH expression by miR-17-5p involved pathogenesis of AS. Our topics chose transgenic mice and patients fibroblasts. Using a variety of techniques to verify that miR-17-5p regulates ANKH expression and promotes ectopic calcification in vivo and in vitro. We attempt to clarify the role of ANKH and miR-17-5p in pathogenesis of AS and find new drug targets for the prevention and treatment of AS.

强直性脊柱炎(ankylosing spondylitis,AS)发病机制未明。ANKH基因(progressive ankylosis gene) 缺陷小鼠的表型，类似于人强直性脊柱炎，但无证据显示患者ANKH发生突变。AS患者MicroRNA Pattern中发现miR-17-5p的表达显著升高。我们通过在线预测及双荧光报告基因实验，基本判定miR-17-5p的靶基因是ANKH。同时，检测患者棘上韧带成纤维细胞，发现miR-17-5p表达增高，ANKH基因的转录水平下调。由此我们推测：AS发病过程中， miR-17-5p表达升高，可能通过下调ANKH的表达，参与AS发病。本课题选用转基因动物以及患者成纤维细胞，分别在患者细胞，转基因动物体内、外验证miR-17-5p调控 ANKH并参与异位钙化的发生，以其系统探讨miR-17-5p→ANKH的调控规律，阐明其在AS发病过程中的作用。

强直性脊柱炎(ankylosing spondylitis,AS)发病机制未明。ANKH 基因(progressive ankylosis gene) 缺陷小鼠的表型，类似于人强直性脊柱炎，但无证据显示患者ANKH 发生突变。本课题以强直性脊柱炎患者的组织及血液为样本，检测这些AS患者韧带、关节囊miR-17-5p、ANKH 表达水平，提取RNA，检测miR-17-5p 表达水平。探讨miR-17-5p在正常和强直性脊柱炎（AS）患者关节囊组织中的表达水平差异，同时检测患者棘上韧带成纤维细胞，发现miR-17-5p 表达增高，ANKH 基因的转录水平下调。进而研究miR-17-5p调控ANKH的表达影响成纤维细胞异位骨化的细胞生物学机制，进一步通过在线预测及双荧光报告基因实验，基本判定miR-17-5p 的靶基因是ANKH。选用转基因动物以及患者成纤维细胞，分别在患者细胞，转基因动物体内、外验证miR-17-5p调控 ANKH 并参与异位钙化的发生，以其系统探讨miR-17-5p→ANKH 的调控规律，阐明其在AS 发病过程中的作用。为强直性脊柱炎疾病探寻新的治疗靶点并提供新的理论依据。