Quantitative Proteomic Analysis of Rat Condylar Chondrocytes during Postnatal Development.

Li Jiang, Yin-yin Xie, Li Wei, Qi Zhou, Xing Shen, Yiming Gao, Xinquan Jiang
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Abstract

OBJECTIVE To investigate differentially expressed proteins in rat mandibular condylar cartilage (MCC) chondrocytes caused by initial mastication for short postnatal periods. METHODS Four groups of protein samples were extracted from primary cultured rat MCC chondrocytes, harvested from eigthy postnatal SD rats aged 1,7,14 and 28 days, with twenty in each group. Total proteins were labelled with isobaric tags for relative and absolute quantification (iTRAQ) reagents. Two-dimensional nano-high-performance liquid chromatography (HPLC) and matrix-assisted laser desorption ionization-time-of-flight/ time-of-flight (MALDI-TOF/TOF) mass spectrometry analysis with iTRAQ technique were performed. All data were analysed by MASCOT software with the SWISSPROT protein database. Furthermore, bioinformatics and statistical analysis were performed to classify their cellular components, biological processes, molecular functions and metabolic pathway by the PANTHER database. RESULTS In total, 137 differentially expressed proteins were identified during MCC growth and were assigned to one or more cellular components. According to the PANTHER analysis, a significant proportion of proteins are involved in the metabolic process, cellular process, biological regulation, developmental process and response to stimulus. The most extensive molecular function was 43% in catalytic activity. In addition, it was found that proteins in MCC chondrocytes change markedly on the growth stage of eruption of the teeth. CONCLUSION This study provides an integrated perspective of molecular mechanisms regulating early normal postnatal growth and development of rat MCC at the protein level.
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大鼠出生后髁突软骨细胞的定量蛋白质组学分析。
目的探讨产后短时间初始咀嚼对大鼠下颌髁突软骨细胞差异表达蛋白的影响。方法从1、7、14、28日龄SD大鼠8只原代培养大鼠MCC软骨细胞中提取蛋白样品,每组20只。总蛋白用等压标记进行相对定量和绝对定量(iTRAQ)试剂标记。采用iTRAQ技术进行二维纳米高效液相色谱(HPLC)和基质辅助激光解吸电离飞行时间/飞行时间(MALDI-TOF/TOF)质谱分析。所有数据用MASCOT软件与SWISSPROT蛋白数据库进行分析。利用PANTHER数据库对其细胞成分、生物学过程、分子功能和代谢途径进行生物信息学和统计学分析。结果在MCC生长过程中共鉴定出137个差异表达蛋白,这些蛋白被分配到一个或多个细胞成分中。根据PANTHER分析,有相当比例的蛋白质参与代谢过程、细胞过程、生物调节、发育过程和对刺激的反应。最广泛的分子功能为43%的催化活性。此外,我们还发现在萌牙生长阶段,MCC软骨细胞中的蛋白发生了显著的变化。结论本研究从蛋白水平上对大鼠MCC早期正常出生后生长发育的分子机制进行了综合研究。
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