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牛关节软骨中微小RNA表达谱及其对机械转导的影响

Profiling microRNA expression in bovine articular cartilage and implications for mechanotransduction.

作者信息

Dunn Walter, DuRaine Grayson, Reddi A Hari

机构信息

Lawrence Ellison Center for Tissue Regeneration and Repair, Department of Orthopaedic Surgery, University of California, Davis, Sacramento, CA 95817, USA.

出版信息

Arthritis Rheum. 2009 Aug;60(8):2333-9. doi: 10.1002/art.24678.

Abstract

OBJECTIVE

Articular cartilage is an avascular tissue with precise polarity and organization comprising 3 distinct functional zones: the surface, middle, and deep zones. Each zone has a different gene expression pattern that plays a specific role in articular cartilage development and maintenance. MicroRNA (miRNA) are small noncoding gene products that play an important regulatory role in determining cell differentiation and function. The purpose of this study was to test our hypothesis that miRNA expression profiles in the different articular cartilage zones as well as between regions subjected to different levels of weight-bearing stresses are unique.

METHODS

Using an miRNA microarray approach in conjunction with quantitative reverse transcription-polymerase chain reaction, we identified miRNA in bovine articular cartilage that were differentially expressed in the different functional zones and in the anterior weight-bearing and posterior non-weight-bearing regions of the medial femoral condyle (M1 and M4, respectively).

RESULTS

We identified miRNA-221 and miR-222 as part of a subset of differentially expressed miRNA that were up-regulated in articular cartilage in the anterior, M1, greater weight-bearing location. Additionally, miR-126, miR-145, and miR-335 were down-regulated in monolayers of tissue-cultured chondrocytes as compared with levels determined directly from intact native cartilage.

CONCLUSION

In conclusion, miR-222 expression patterns in articular cartilage are higher in the weight-bearing anterior medial condyle as compared with the posterior non-weight-bearing medial condyle. Thus, miR-222 might be a potential regulator of an articular cartilage mechanotransduction pathway. These data implicate miRNA in the maintenance of articular cartilage homeostasis and are therefore targets for articular cartilage tissue engineering and regenerative medicine.

摘要

目的

关节软骨是一种无血管组织,具有精确的极性和组织结构,由3个不同的功能区组成:表层、中层和深层。每个区域都有不同的基因表达模式,在关节软骨的发育和维持中发挥特定作用。微小RNA(miRNA)是小的非编码基因产物,在决定细胞分化和功能方面发挥重要的调节作用。本研究的目的是验证我们的假设,即不同关节软骨区域以及承受不同程度负重应力区域之间的miRNA表达谱是独特的。

方法

我们采用miRNA微阵列方法结合定量逆转录-聚合酶链反应,鉴定了牛关节软骨中在不同功能区以及股骨内侧髁的前侧负重区和后侧非负重区(分别为M1和M4)中差异表达的miRNA。

结果

我们鉴定出miRNA-221和miR-222是差异表达miRNA子集的一部分,它们在关节软骨的前侧(M1)、负重更大的位置上调。此外,与直接从完整天然软骨中测定的水平相比,miR-126、miR-145和miR-335在组织培养软骨细胞单层中下调。

结论

总之,与后侧非负重的内侧髁相比,关节软骨中miR-222在负重的前内侧髁中的表达模式更高。因此,miR-222可能是关节软骨机械转导途径的潜在调节因子。这些数据表明miRNA参与关节软骨内环境稳态的维持,因此是关节软骨组织工程和再生医学的靶点。

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