CAJ | 학술논문

目的：通过建立的大鼠髁突软骨细胞发育过程中蛋白表达差异谱，分析可能参与的信号通路，为进一步认识髁突软骨生理性及病理性生长改建的信号调控机制提供相关信息。方法：收获出生后1、7、14、28 d共4组SD大鼠髁突软骨细胞，甲苯胺蓝染色、Ⅱ型胶原免疫组化染色鉴定软骨细胞。提取各组软骨细胞总蛋白，采用iTRAQ标记定量蛋白，2D nano-HPLC和基质辅助激光解吸/电离串联飞行时间质谱（MALDI-TOF-TOF），获取出生后大鼠髁突软骨细胞发育过程中差异蛋白的表达谱，所得数据用MASCOT软件处理，筛选样本之间有意义的差异蛋白，运用GO法及David软件进行KEGG信号通路分析。结果：共鉴定137种具有可信度表达的蛋白，有44种蛋白参与至少27条KEGG信号通路，其中ECM-受体相互作用、焦点粘连、actin骨架调节、Ca2+信号通路、血管平滑肌收缩、GnRH信号通路、肌醇三磷酸代谢、磷脂酰肌醇信号系统以及核糖体等信号通路具有统计学意义。结论：在大鼠髁突软骨发育中，各信号通路蛋白在时间、空间上差异性表达，共同形成复杂的信号传递网络。
목적：통과건립적대서과돌연골세포발육과정중단백표체차이보，분석가능삼여적신호통로，위진일보인식과돌연골생이성급병이성생장개건적신호조공궤제제공상관신식。방법：수획출생후1、7、14、28 d공4조SD대서과돌연골세포，갑분알람염색、Ⅱ형효원면역조화염색감정연골세포。제취각조연골세포총단백，채용iTRAQ표기정량단백，2D nano-HPLC화기질보조격광해흡/전리천련비행시간질보（MALDI-TOF-TOF），획취출생후대서과돌연골세포발육과정중차이단백적표체보，소득수거용MASCOT연건처리，사선양본지간유의의적차이단백，운용GO법급David연건진행KEGG신호통로분석。결과：공감정137충구유가신도표체적단백，유44충단백삼여지소27조KEGG신호통로，기중ECM-수체상호작용、초점점련、actin골가조절、Ca2+신호통로、혈관평활기수축、GnRH신호통로、기순삼린산대사、린지선기순신호계통이급핵당체등신호통로구유통계학의의。결론：재대서과돌연골발육중，각신호통로단백재시간、공간상차이성표체，공동형성복잡적신호전체망락。
Objective: To analyze related-signal pathways regulating proliferation and differentiation of rat mandibular condylar chondrocytes. Methods:The 1-, 7-, 14-, and 28-day-old SD rats′condylar chondrocytes were harvested in vitro, identified by toluidine blue stain and type II collagen immunohistochemistry reaction. After extracting from rat condyle in each group, total proteins were labeled with iTRAQ reagents and resolved using 2D nano-high performance liquid chro-matography (HPLC). Protein spots were then identified by mass spectrometry. Using matrix-assisted laser desorption ioniza-tion-time-of-flight/time-of-flight technology (MALDI-TOF/TOF) to quantitative analysis the differential protein expression of rat condylar chondrocytes during their development. All data was normalized by MASCOT software to search useful differ-entially expressed proteins, then analyzed by Gene Ontology and David software, KEGG pathway database. Results: 137 differentially reliability proteins were identified, of which 44 proteins were found to be involved in at least 27 signalling subpathways. There were statistically significant differences in ECM-receptor interaction, focal adhesion, regulation of actin cytoskeleton, calcium signaling pathway, vascular smooth muscle contraction, GnRH signal pathway, inositol phosphate metabolism, phosphatidylinositol signaling system and ribosome. Conclusion: This study indicated that during rat condylar development, differential proteins involved in signal transduction pathways expressed spatially and temporally.