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  

Effect of pearl powder on the proliferation,differentiation,and mineralization of MC3T3-E1 cells and the expression of bone related genes in the cells

作　　者：王凯[1] 魏博[1] 谭佳妮[1] 季晖[1] 

机构地区：[1]中国药科大学药理教研室,南京210009

出　　处：《中国骨质疏松杂志》2014年第6期666-672,共7页Chinese Journal of Osteoporosis

摘　　要：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。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.

关 键 词：珍珠粉 MC3T3-E1细胞 成骨活性 

分 类 号：R965[医药卫生—药理学]