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首页> 外文期刊>Osteoarthritis and cartilage >Proteome characterization of human articular chondrocytes leads to novel insights in the function of small heat-shock proteins in chondrocyte homeostasis.
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Proteome characterization of human articular chondrocytes leads to novel insights in the function of small heat-shock proteins in chondrocyte homeostasis.

机译：人关节软骨细胞的蛋白质组学表征导致小型热休克蛋白在软骨细胞稳态中的功能的新见解。

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OBJECTIVE: In recent years, studies have been initiated to disclose the proteome of human chondrocytes and cartilage. Despite these studies, comprehensive information of the chondrocyte proteome remains limited. This study aimed to further explore the proteome expressed by human knee chondrocytes, and to study the functional aspects of heat-shock protein 27 (HSP27), a protein related to the previously described alphaBcrystallin, in chondrocyte biology. METHODS: Chondrocytes isolated from human knee articular cartilage were cultured in a three-dimensional alginate culture system. To simplify the protein mixtures, proteins extracted from chondrocyte cell lysates were fractionated based on hydrophobicity and molecular weight. Proteins were digested and the resulting peptides were separated and identified by an on-line two-dimensional (2-D) nanoliquid chromatography (nanoLC)-system coupled to a quadrupole time-of-flight (Qq-TOF) mass spectrometer. Differential expression analysis of HSP27 was performed by Western Blotting and quantitative polymerase chain reaction (QPCR). The effects of HSP27 on chondrocyte biology were explored by suppression of HSP27 expression induced by RNA interference (RNAi). RESULTS: In this study, we identified proteins with unknown functions together with membrane proteins, transcription factors and other low abundant proteins, which have not yet been described in chondrocytes. Based on previous knowledge on the related protein alphaBcrystallin, we selected HSP27 from the chondrocyte proteome database. Differential expression analysis revealed a decreased expression of HSP27 in Osteoarthritic (OA) chondrocytes. RNAi experiments revealed that HSP27 is involved in interleukin-1beta (IL-1beta) induced IL-6 secretion. CONCLUSION: These findings highlight that small HSPs, especially HSP27, play a prominent role in the maintenance of human articular chondrocyte homeostasis.
机译：目的：近年来，已经开始进行研究以揭示人软骨细胞和软骨的蛋白质组。尽管进行了这些研究，但软骨细胞蛋白质组的综合信息仍然有限。这项研究旨在进一步探索人膝软骨细胞表达的蛋白质组，并研究软骨细胞生物学中热休克蛋白27（HSP27）的功能方面，该蛋白与先前描述的alphaBcrystallin有关。方法：从人膝关节软骨分离的软骨细胞在三维藻酸盐培养系统中培养。为了简化蛋白质混合物，将从软骨细胞裂解物中提取的蛋白质根据疏水性和分子量进行分级分离。消化蛋白质，并通过与二维四极杆飞行时间（Qq-TOF）质谱仪耦合的在线二维（2-D）纳米液相色谱（nanoLC）系统分离和鉴定所得肽。通过蛋白质印迹和定量聚合酶链反应（QPCR）进行HSP27的差异表达分析。通过抑制RNA干扰（RNAi）诱导的HSP27表达，探索了HSP27对软骨细胞生物学的影响。结果：在这项研究中，我们鉴定了功能未知的蛋白质以及膜蛋白质，转录因子和其他低丰度蛋白质，而软骨细胞中尚未对此进行描述。基于对相关蛋白alphaBcrystallin的先前了解，我们从软骨细胞蛋白质组数据库中选择了HSP27。差异表达分析表明，骨关节炎（OA）软骨细胞中HSP27的表达降低。 RNAi实验表明，HSP27参与白介素-1β（IL-1beta）诱导的IL-6分泌。结论：这些发现表明，小的HSP，尤其是HSP27，在维持人类关节软骨细胞稳态中起着重要作用。

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