卵黄高磷蛋白磷酸肽及其钙螯合物在双细胞共培养体系中对成骨细胞分化的促进作用

Phosvitin Phosphopeptide and Its Calcium Chelate Promote Osteoblast Differentiation in A Double Cell Co-culture System

摘要:

研究了卵黄高磷蛋白磷酸肽（Phosvitin Phosphopeptide，PPP）及其钙螯合物（PPP-Ca）在成骨细胞（MC3T3-E1）和破骨前体细胞（RAW264.7）共培养体系中对成骨细胞分化的调节作用。对细胞毒性、碱性磷酸酶（AKP）、抗酒石酸酸性磷酸酶（TRAP）的活性和TRAP染色进行分析；用RT-PCR技术进一步探究成骨细胞RANKL/OPG通路相关蛋白的mRNA表达情况。结果发现，PPP和PPP-Ca可以使MC3T3-E1体系中的AKP活性分别增加9.5%和12.7%；PPP和PPP-Ca的加入可以使MC3T3-E1体系中OPG基因mRNA的表达量从0.92分别提升至1.25和1.39、使RANKL基因mRNA的表达量从1.00分别提升至1.23和1.45。该研究结果表明，PPP和PPP-Ca可有效促进成骨细胞的分化。实验结果为进一步探索磷酸肽在多细胞模型体系中的作用提供了研究基础，同时为磷酸肽的功能活性开发提供理论依据。

Abstract:

The regulatory effects of phosvitin phosphopeptide (PPP) and its calcium chelate (PPP-Ca) on osteoblast differentiation in the co-culture system of osteoblast (MC3T3-E1) and osteoclast precursor (RAW264.7) were investigated. Cytotoxicity, alkaline phosphatase (AKP) activity, tartrate-resistant acid phosphatase (TRAP) activity and TRAP staining were analyzed. The mRNA expressions of RANKL/OPG pathway-related proteins in osteoblasts were further studied by RT-PCR. The results showed that PPP and PPP-Ca increased the AKP activity in the MC3T3-E1 system by 9.5% and 12.7%, respectively. In the MC3T3-E1 system, PPP and PPP-ca increased the mRNA expression levels from 0.92 to 1.25 and 1.39, respectively, for OPG, and from 1.00 to 1.23 and 1.45, respectively for RANKL. The results of this study showed that PPP and PPP-ca could effectively promote the differentiation of osteoblasts. The experimental results provide a research basis for further exploration of the role of phosphopeptides in the multi-cell model system and the development of functional activity of phosphopeptides.