Department of Biomedical Laboratory Science, School of Medicine, Eulji University, Daejeon 34824, Republic of Korea.
Department of Integrated Material's Development, CHA Meditech Co., Ltd, Daejeon 34025, Republic of Korea.
Differentiation. 2018 May-Jun;101:1-7. doi: 10.1016/j.diff.2018.03.001. Epub 2018 Mar 15.
Vitamin C-linker-conjugated Ala-His-Lys tripeptide (Vit C-AHK) is a derivative of Vitamin C-conjugated tripeptides, which were originally developed as a component of a product for collagen synthesis enhancement or human dermal fibroblast growth. Here, we investigated the effect of Vit C-AHK on bone morphogenetic protein (BMP)-2-induced osteoblast differentiation in a cell culture model. Vit C-AHK enhanced proliferation of C2C12 cells and induction of BMP-2-induced alkaline phosphatase, a typical marker of osteoblast differentiation. Vit C-AHK also stimulated the phosphorylation and translocation of Smad1/5/8 to the nucleus and phosphorylation of mitogen-activated protein kinases (MAPKs) including ERK1/2 and p38. In addition, Vit C-AHK enhanced the BMP-2-induced mRNA expression of osteoblast differentiation-related genes such as ALP, BMP-2, Osteocalcin, and Runx2. Our results suggest that Vit C-AHK exerts an enhancing effect on osteoblast proliferation and differentiation through activation of Smad1/5/8 and MAPK ERK1/2 and p38 signaling and without significant cytotoxicity. These results provide important data for the development of peptide-based bone-regenerative agents and treatment of bone-related disorders.
维生素 C-连接子-缀合的 Ala-His-Lys 三肽 (Vit C-AHK) 是维生素 C-缀合三肽的衍生物，最初被开发为增强胶原蛋白合成或人真皮成纤维细胞生长的产品的一部分。在这里，我们研究了 Vit C-AHK 在细胞培养模型中对骨形态发生蛋白 (BMP)-2 诱导的成骨细胞分化的影响。Vit C-AHK 增强了 C2C12 细胞的增殖和 BMP-2 诱导的碱性磷酸酶的诱导，碱性磷酸酶是成骨细胞分化的典型标志物。Vit C-AHK 还刺激 Smad1/5/8 的磷酸化和向核内易位以及包括 ERK1/2 和 p38 在内的丝裂原激活蛋白激酶 (MAPK) 的磷酸化。此外，Vit C-AHK 增强了 BMP-2 诱导的成骨细胞分化相关基因如 ALP、BMP-2、骨钙素和 Runx2 的 mRNA 表达。我们的结果表明，Vit C-AHK 通过激活 Smad1/5/8 和 MAPK ERK1/2 和 p38 信号通路，增强成骨细胞的增殖和分化，而没有明显的细胞毒性。这些结果为基于肽的骨再生剂的开发和骨相关疾病的治疗提供了重要数据。
