620nm非相干红光对大鼠骨髓间充质干细胞的光生物调节作用.pdfVIP

（3 ）细胞在光照下有明显的增殖，总mTOR 的水平没有明显变化，但mTOR 的 磷酸化水平增高； （4 ）受光照的细胞有更多的钙结节生成，Cry2、Per2 、Clock 的表达水平有不同 程度的上升。 结论：（1）密度梯度离心法结合贴壁法可获得均一、稳定、可靠的间充质干细胞； （2 ）低能量非相干光可以促进普通培养基中的间充质干细胞增殖，不能促进其 向成骨分化，但可促进成骨诱导基中的间充质干细胞成骨分化并使其增殖速度减 缓； （3 ）低能量非相干光可促进间充质干细胞增殖并激活mTOR 信号通路； （4 ）昼夜节律基因可能参与调控低能量非相干光诱导的间充质干细胞成骨分化。 关键词：骨髓间充质干细胞；非相干光；光生物调节作用；低能量光照治疗；增 殖；成骨分化；信号通路；昼夜节律 2 Abstract Object: (1) To obtain purity and stable bone marrow mesenchymal stem cells (MSCs) culutured in vitro; (2) To investigate the effect of red light emitted from light-emitting diode (LED) on bone marrow MSCs with or without osteogenic supplements; (3) To investigate the activation of mTOR in photoinduced proliferation of rat MSCs; (4) To investigate the expression activation of mTOR in photoinduced proliferation of rat MSCs; Method: (1) MSCs were isolated by density gradient centrifugation combined adherence culture, observed under optical microscope and detected by flow cytometry. The cells were induced into osteoblasts, fat cells and chondrogenic cells, and identified by von kossa staining, oil red O staining and alcian blue staining; (2) MSCs with and without osteogenic supplements both were divided into four 2 groups and each group was irradiated at dose of 0, 1, 2, and 4J/cm . Cellular proliferation was evaluated by using CCK-8 and 5-ethynyl-2′-deoxyuridine (EdU) fluorescence staining. The alkaline phosphatase (ALP) activity, mineralization formation, and expression of osteoblast master genes (Col1 α1, Alpl , Bglap and Runx2) were monitored as indicators of MSCs’ differentiation toward osteoblasts. In groups withou