Identification of key miRNAs in unilateral mastication induced disruption of cartilage homeostasis
Identification of key miRNAs in unilateral mastication induced disruption of cartilage homeostasis
复制标题
单侧咀嚼引起的软骨稳态破坏中关键 miRNA 的鉴定
DOI：
10.1111/odi.14504
复制
发表时间：
2023
期刊：
影响因子：
3.8
通讯作者：
Mengrui Wu
中科院分区：
文献类型：
--
作者：
Mengjie Wu;Xuebin Wang;Jing Shuai;Liquan Deng;Haiping Lu;Yiqun Zhou;Mengrui Wu
目的本研究鉴定了异常应力下颞下颌关节软骨分化的潜在关键miRNA。材料与方法Sprague-Dawley大鼠随机分为对照组和实验性单侧咀嚼（EUM）组。显微CT检测骨微结构参数，免疫组化检测成纤维细胞生长因子-1（FGF-1）和基质金属蛋白酶-1（MMP-1）表达。在4周和8周EUM时通过miRNA微阵列筛选双侧髁突软骨差异表达的miRNA，然后使用定量逆转录PCR进一步验证。软骨细胞中五种差异表达的miRNA的过表达是通过抑制miRNA模拟物来触发的。Western blotting检测MMP-13、Col-II、OPN和Runx 2的表达。结果EUM后2 ~ 6周，右侧髁突FGF-1和MMP-1的表达逐渐增加。共发现20个差异表达的miRNAs受EUM调控，它们与细胞增殖、侵袭和成骨细胞分化途径有关。miR-148 a-3 p和miR-1 - 3 p的过表达导致Col-II下调，而MMP-13和Runx 2通过诱导营养不良分化或IL-1β刺激而上调。这些结果表明miR-148 a-3 p和miR-1 - 3 p促进软骨分化。结论发现了几个关键的miRNA与软骨分化有关，这为软骨稳态的致病机制提供了新的见解。
ObjectiveThe present study identified potentially pivotal miRNAs contributing to chondrogenic differentiation in temporomandibular joint suffering abnormal stress.Materials and MethodsSprague–Dawley rats were randomly divided into control and experimental unilateral mastication (EUM) group. Bone micro‐structure parameters was detected by micro‐CT, and FGF‐1 and MMP‐1 expression was examined by immunohistochemistry. Differentially expressed miRNAs of bilateral condyle cartilage were screened via miRNA microarray at 4‐ and 8‐week EUM, then further verified using quantitative reverse‐transcription PCR. Over‐expression of five differentially expressed miRNAs in chondrocytes was triggered by transfecting miRNA mimics. The expression of MMP‐13, Col‐II, OPN, and Runx2 was verified by western blotting.ResultsExpressions of FGF‐1 and MMP‐1 in right condyles gradually increased from 2 to 6 weeks after EUM. A total of 20 differentially expressed miRNAs were regulated by EUM, which related to cell proliferation, invasion, and osteoblast differentiation pathways. The over‐expression of miR‐148a‐3p and miR‐1‐3p led to down‐regulation of Col‐II, while MMP‐13 and Runx2 were up‐regulated by induction of hypotrophic differentiation or IL‐1β stimulation. These findings suggested that miR‐148a‐3p and miR‐1‐3p promote chondrogenic differentiation.ConclusionsSeveral pivotal miRNAs were found to be related to chondrogenic differentiation, which provides novel insight into pathogenic mechanisms of cartilage homeostasis.