CAJ | 학술논문

目的观察解聚素样金属蛋白酶10（A disintegrin and metalloprotease 10，ADAM10）在小鼠颅面部骨骼膜发育过程中的表达变化。方法以野生型C57BL/6小鼠为实验对象，阿辛蓝-茜素红染色显示小鼠膜内成骨区域，结合免疫组织荧光，检测ADAM10在骨组织中的表达。三标免疫荧光观察ADAM10在MC3T3细胞系中亚细胞定位。蛋白免疫印迹检测ADAM10在小鼠出生前后的表达量变化。结果组织阿辛蓝茜素红染色显示小鼠前颅骨、鼻旁软骨、上颌骨、腭板和下颌骨成骨活跃，并结合免疫组织荧光检测，发现ADAM10在小鼠前颌骨、鼻旁软骨、上颌骨、腭板和下颌骨广泛表达，且主要表达在成骨活跃的区域。 MC3T3细胞三标免疫荧光检测进一步定位ADAM10广泛分布在胞浆中，在质膜和细胞核附近表达高；其胞核附近的高表达信号与高尔基体共标，提示其可能在高尔基体中加工后至细胞膜发挥功能。同时，蛋白免疫印迹结果证实，ADAM10在小鼠出生前后表达高，成年后表达明显降低。结论 ADAM10广泛表达于小鼠早期颅面部膜内成骨活跃区域，提示其可能调控小鼠颅面部膜内成骨过程。
목적관찰해취소양금속단백매10（A disintegrin and metalloprotease 10，ADAM10）재소서로면부골격막발육과정중적표체변화。방법이야생형C57BL/6소서위실험대상，아신람-천소홍염색현시소서막내성골구역，결합면역조직형광，검측ADAM10재골조직중적표체。삼표면역형광관찰ADAM10재MC3T3세포계중아세포정위。단백면역인적검측ADAM10재소서출생전후적표체량변화。결과조직아신람천소홍염색현시소서전로골、비방연골、상합골、악판화하합골성골활약，병결합면역조직형광검측，발현ADAM10재소서전합골、비방연골、상합골、악판화하합골엄범표체，차주요표체재성골활약적구역。 MC3T3세포삼표면역형광검측진일보정위ADAM10엄범분포재포장중，재질막화세포핵부근표체고；기포핵부근적고표체신호여고이기체공표，제시기가능재고이기체중가공후지세포막발휘공능。동시，단백면역인적결과증실，ADAM10재소서출생전후표체고，성년후표체명현강저。결론 ADAM10엄범표체우소서조기로면부막내성골활약구역，제시기가능조공소서로면부막내성골과정。
Objective To investigate the expression of ADAM10 during craniomaxillofacial ossification of mice. Methods Alcian blue and Alizarin red staining were used to detect the intramembranous ossification areas during craniomaxillofacial development of wild-type C57BL/6 mice. Expression of ADAM10 in mice craniomaxillofacial bone was analyzed by immunohistochemistry. The subcellular localization of ADAM10 in MC3T3 was observed by double immunofluorescent staining and the expression of ADAM10 in mice mandibular was determined by western blot. Results Skull, nasal cartilage, maxilla, palate plate and mandibular of mice displayed intramembranous ossification and expression of ADAM10 showed strong singal in these regions, especially the regions undergoing intramembranous ossification. In MC3T3, ADAM10 distributed in cytoplasm, especially in cell membrance and cell nucleus, and the high expression signal of ADAM10 near cell nucleus was co-localized with GM130, a marker of Golgi, indicating ADAM10 decorated in Golgi before transported to the cell membrance. The expression of ADAM10 in mice mandibular was significantly down-regulated when grown up. Conclusion ADAM10 is widely expressed in mice intramembranous ossification regions, providing the evidence of its regulation in intramembranous ossification of mice.