Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical & Health Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.
Curr Pharm Des. 2018;24(25):2993-3001. doi: 10.2174/1381612824666180816093227.
Amelogenins are enamel matrix proteins that play crucial roles in enamel formation. Previous studies have indicated that amelogenin and amelogenin C-terminal peptides have cell-signaling functions. Recently, adipocyte-derived mesenchymal stem cells (ADSCs) have received attention as a potential source of stem cells for use in regeneration therapy. In this study, we examined the effects of human full-length amelogenin (rh174) and amelogenin C-terminal peptide (amgCP) on the proliferation of ADSCs. ADSCs were cultured in the presence of amgCP or rh174. Cell proliferation was analyzed using BrdU immunoassay and MTS assay. Cell migration was evaluated by ELISA. The MAPK-ERK pathway was examined by phospho-p44/42 MAPK (Thr202/Tyr204) sandwich ELISA and western blotting. A specific MAPK inhibitor, U0126, was used to block ERK activity. ADSC proliferation and migration were significantly (P < 0.05) increased in the presence of rh174 or amgCP compared to non-treated control cells. The increased proliferation of ADSCs induced by rh174 or amgCP was significantly (P < 0.05) inhibited in the presence of 2 µg/ml U0126. The pERK/tERK ratio was significantly (P < 0.05) increased upon treatment with rh174 or amgCP compared to non-treated ADSCs, while this increase was significantly (P < 0.05) suppressed by the addition of U0126. Similar results were found by western blot analysis. In conclusion, amgCP and rh174 increase ADSC proliferation via the MAPK-ERK signaling pathway, and ADSCs may be useful for tissue regeneration in the orofacial region.
釉原蛋白是釉基质蛋白，在釉质形成中起着至关重要的作用。先前的研究表明釉原蛋白和釉原蛋白 C 端肽具有细胞信号功能。最近，脂肪细胞源性间充质干细胞（ADSCs）作为再生治疗中潜在的干细胞来源受到关注。在这项研究中，我们研究了人全长釉原蛋白（rh174）和釉原蛋白 C 端肽（amgCP）对 ADSC 增殖的影响。在存在 amgCP 或 rh174 的情况下培养 ADSC。通过 BrdU 免疫测定和 MTS 测定分析细胞增殖。通过 ELISA 评估细胞迁移。通过磷酸化 p44/42 MAPK（Thr202/Tyr204）夹心 ELISA 和 Western blot 检查 MAPK-ERK 途径。使用特定的 MAPK 抑制剂 U0126 阻断 ERK 活性。与未处理的对照细胞相比，rh174 或 amgCP 的存在显著（P <0.05）增加了 ADSC 的增殖和迁移。rh174 或 amgCP 诱导的 ADSC 增殖显著（P <0.05）被 2μg/ml U0126 抑制。与未处理的 ADSC 相比，rh174 或 amgCP 处理后 pERK/tERK 比值显著（P <0.05）增加，而 U0126 的添加显著（P <0.05）抑制了这种增加。Western blot 分析也得到了类似的结果。总之，amgCP 和 rh174 通过 MAPK-ERK 信号通路增加 ADSC 增殖，ADSCs 可能可用于口腔颌面部组织再生。
