珍珠粉对 MC3T3-E1细胞增殖、分化、矿化及骨相关基因表达的影响

摘要

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.%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。