CAJ | 학술논문

目的：研究珍珠粉对MC3T3-E1细胞成骨增殖、分化、间质矿化及凋亡的影响，并初步探讨其作用机制。方法在体外培养的MC3T3-E1细胞中分别加入珍珠粉和低钙珍珠粉，采用MTT法及碱性磷酸酶（Alkaline phosphatase，ALP）试剂盒检测其对MC3T3-E1细胞增殖及分化的影响；Von Kossa染色法观察矿化结节的形成；RT-PCR检测细胞中转录因子（ Runt-related transcription factor II，Runx2）、骨钙素（Osteocalcin，OC）、骨保护因子（Osteoprotegerin， OPG）及核因子-κB受体活化因子配体（Receptor activator of NK-κB ligand， RANKL）mRNA的表达；流式细胞术检测血清饥饿诱导的细胞凋亡。结果珍珠粉及低钙珍珠粉在一定浓度范围内，能促进MC3T3-E1细胞成熟期的ALP合成分泌和矿化结节的形成，并对Runx2及OC基因表达具有明显促进作用，可降低RANKL/OPG的比率，明显改善血清饥饿诱导的成骨细胞凋亡。结论珍珠粉及低钙珍珠粉对MC3T3-E1细胞分化成熟过程有显著的促进作用，前者作用显著强于后者，说明珍珠粉中钙成分对MC3T3-E1细胞分化成熟起到主要作用。
목적：연구진주분대MC3T3-E1세포성골증식、분화、간질광화급조망적영향，병초보탐토기작용궤제。방법재체외배양적MC3T3-E1세포중분별가입진주분화저개진주분，채용MTT법급감성린산매（Alkaline phosphatase，ALP）시제합검측기대MC3T3-E1세포증식급분화적영향；Von Kossa염색법관찰광화결절적형성；RT-PCR검측세포중전록인자（ Runt-related transcription factor II，Runx2）、골개소（Osteocalcin，OC）、골보호인자（Osteoprotegerin， OPG）급핵인자-κB수체활화인자배체（Receptor activator of NK-κB ligand， RANKL）mRNA적표체；류식세포술검측혈청기아유도적세포조망。결과진주분급저개진주분재일정농도범위내，능촉진MC3T3-E1세포성숙기적ALP합성분비화광화결절적형성，병대Runx2급OC기인표체구유명현촉진작용，가강저RANKL/OPG적비솔，명현개선혈청기아유도적성골세포조망。결론진주분급저개진주분대MC3T3-E1세포분화성숙과정유현저적촉진작용，전자작용현저강우후자，설명진주분중개성분대MC3T3-E1세포분화성숙기도주요작용。
Objective To investigate the effect of pearl powder on the osteogenic proliferation, differentiation, interstitial mineralization, and apoptosis in MC3T3-E1 cells, and to explore the potential mechanisms.Methods MC3T3-E1 cells were cultured with pearl powder or low-calcium pearl powder in vitro.The osteogenic proliferation was determined using MTT method. And the differentiation was determined using alkaline phosphatase ( ALP) kit.The formation of mineralized nodules was observed using von Kossa staining. The mRNA expression of Runt-related transcription factor II ( Runx2 ) , osteocalcin ( OC ) , osteoprotegerin ( OPG ) , and receptor activator of NK-κB ligand ( RANKL ) was detected using RT-PCR.The cell apoptosis induced by serum deprivation was detected using flow cytometry.Results In a certain range of concentration, pearl powder and low-calcium pearl powder could significantly increase the secretion of ALP and the formation of mineralized nodules, and markedly improve the mRNA expression of Runx2 and OC in MC3T3-E1cells.Furthermore, pearl powder and low calcium pearl powder could significantly decrease the RANKL/OPG ratio, and ameliorate the apoptosis of the osteoblasts induced by serum deprivation. Conclusion Both pearl powder and low calcium pearl powder can promote the differentiation and maturation in MC3T3-E1cells. And the effect of pearl powder is much stronger, suggesting a critical role of calcium contained in pearl powder in regulating the osteogenic proliferation and differentiation in MC3T3-E1cells.