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血清 miR-214 作为一种新的强直性脊柱炎生物标志物。

Serum miR-214 as a novel biomarker for ankylosing spondylitis.

机构信息

Department of Rheumatology, Research Institute of Medical Sciences, Chonnam National University Medical School and Hospital & College of Nursing of Chonnam National University, Gwangju, Korea.

Department of Rheumatology, Research Institute of Medical Sciences, Chonnam National University Medical School and Hospital, Gwangju, Korea.

出版信息

Int J Rheum Dis. 2019 Jul;22(7):1196-1201. doi: 10.1111/1756-185X.13475. Epub 2019 Feb 6.

Abstract

OBJECTIVE

Serum microRNA (miR) in ankylosing spondylitis (AS) patients has been rarely identified. The objective of this study was to find AS-specific miR in sera of patients with AS.

METHODS

Total RNAs were isolated from whole sera of patients with AS, patients with rheumatoid arthritis (RA), and healthy controls (HC) using miRNeasy Serum/Plasma Kit. The presence of miR was assayed using Agilent 2100 Bioanalyzer Small RNA assay. Each RNA sample was used for miR microarray. To verify microarray results, candidate circulating miRs were validated by quantitative polymerase chain reaction (qPCR) using samples from patients with AS (n = 65), patients with RA (n = 25), and HCs (n = 39). Cycle threshold values were converted to copy numbers by drawing a standard curve using a synthetic chemical standard. All clinical values were also evaluated at the time of miR isolation.

RESULTS

A total of 887 miRs were screened for three groups. Lower expression of miR-214 in AS than in HC and RA was observed after normalization of raw data. Finally, lower expression of serum miR-214 was confirmed in AS after validation by qPCR. Correlation analysis showed that the level of miR-214 of AS was significantly associated with Ankylosing Spondylitis Disease Activity Score-C-reactive protein (r = 0.299, P = 0.02). However, other disease-specific variables showed no statistical significance: gender (P = 0.286), peripheral arthritis (P = 0.634), enthesitis (P = 0.464), dacylitis (P = 0.750), psoriasis (P = 0.552), inflammatory bowel disease (P = 0.369), human leukocyte antigen-B27 positivity (P = 0.473), use of non-steroidal anti-inflammatory drugs (P = 0.448), and use of tumor necrosis factor-blocker in the last 3 months (P = 0.505).

CONCLUSION

miR-214 may serve as a noninvasive biomarker for diagnosis of AS. In addition, expression level of miR-214 was associated with disease activity.

摘要

目的

强直性脊柱炎(AS)患者的血清 microRNA(miR)很少被识别。本研究的目的是在 AS 患者的血清中找到 AS 特异性的 miR。

方法

使用 miRNeasy Serum/Plasma Kit 从 AS 患者、类风湿关节炎(RA)患者和健康对照(HC)的全血清中分离总 RNA。使用 Agilent 2100 Bioanalyzer Small RNA assay 检测 miR 的存在。每个 RNA 样本均用于 miR 微阵列。为了验证微阵列结果,使用来自 AS 患者(n=65)、RA 患者(n=25)和 HC(n=39)的样本通过定量聚合酶链反应(qPCR)验证候选循环 miR。通过使用合成化学标准品绘制标准曲线将循环阈值值转换为拷贝数。在 miR 分离时还评估了所有临床值。

结果

对三组进行了总共 887 个 miR 的筛选。在对原始数据进行归一化后,观察到 AS 中的 miR-214 表达低于 HC 和 RA。最后,通过 qPCR 验证,AS 患者血清 miR-214 的表达降低得到确认。相关性分析表明,AS 患者的 miR-214 水平与强直性脊柱炎疾病活动评分-C 反应蛋白(r=0.299,P=0.02)显著相关。然而,其他疾病特异性变量没有统计学意义:性别(P=0.286)、外周关节炎(P=0.634)、附着点炎(P=0.464)、骶髂关节炎(P=0.750)、银屑病(P=0.552)、炎症性肠病(P=0.369)、人类白细胞抗原-B27 阳性(P=0.473)、在过去 3 个月中使用非甾体抗炎药(P=0.448)和使用肿瘤坏死因子阻滞剂(P=0.505)。

结论

miR-214 可作为 AS 诊断的非侵入性生物标志物。此外,miR-214 的表达水平与疾病活动度相关。

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