1.Affiliated Hospital of Xiangnan University;2.湘南学院附属医院
1.Affiliated Hospital of Xiangnan University;2.Affiliated Hospital of Xiangnan University (Clinical College)
目的：探讨多聚磷酸盐（Inorganic polyphosphate , Poly P）对肌腱干细胞增殖和能量代谢的影响。方法：分离培养大鼠肌腱干细胞，采用流式细胞术对其表面分子进行鉴定，然后用不同浓度（0 mM、0.5mM、1.0mM）的多聚磷酸盐处理大鼠肌腱干细胞，分别用CKK-8、细胞周期实验和ATP能量检测试剂盒检测肌腱干细胞增殖能力和能量代谢情况，采用蛋白免疫印迹（Western blot）和免疫荧光检测Collagen I的表达。结果: 流式细胞分析表明成功分离大鼠肌腱干细胞， CKK-8实验结果表明Poly P可以促进肌腱干细胞的增殖，细胞周期实验表明Poly P可以提高细胞G2+S期的百分比，免疫荧光和Western blot结果表明Poly P能够促进肌腱干细胞基质Collagen I的表达。结论: 在体外Poly P可以上调大鼠肌腱干细胞基质合成基因Collagen I的表达和ATP的生成，促进肌腱干细胞的增殖。通过明确Poly P对肌腱干细胞增殖和能量代谢的影响，为治疗肌腱病提供新的思路。
Objective: To investigate the effect of Inorganic polyphosphate (Poly P) on the proliferation and energy metabolism of tendon stem cells. Methods: Rat tendon stem cells were isolated and cultured, with the stem cell surface markers confirmed by flow cytometry analysis. These rat tendon stem cells were treated with different concentrations of Poly P (0, 0.5 mM, 1.0 mM), and their proliferation was tested by CCK8 assay and the cell cycle analysis. Also, the energy metabolism of these tendon stem cells under the treatment of polyphosphate was assessed by studying the levels of ATP production, and the expression of Collagen I was detected by Western blot and immunofluorescence. Results: The successful isolation of the rat tendon stem cells was confirmed by the flow cytometry analysis. We found that Poly P could dose-dependently promote the proliferation of tendon stem cells and increase the proportion of cells in the G2/S phase. Immunofluorescence and Western blot both showed that Poly P could increase the expression of Collagen I in these tendon stem cells. Conclusion: Poly P treatment can promote the proliferation of rat tendon stem cells in vitro. It can also up-regulate the expression of collagen I and increase the ATP production of these tendon stem cells in a dose-dependent manner. Understanding the effect of Poly P on the key features of tendon stem cells may provide valuable information for the treatment of tendinopathy.