Department of Orthopaedics Surgery, Ningbo No. 2 Hospital, Zhejiang, 315010, People's Republic of China.
, 41 Xibei Rd., Haishu, Ningbo, 315000, People's Republic of China.
Mol Cell Biochem. 2018 Jan;438(1-2):175-182. doi: 10.1007/s11010-017-3124-8. Epub 2017 Aug 1.
To investigate the mechanism of recombinant human platelet-derived growth factor-BB (rhPDGF-BB) and human adipose-derived stem cells (hADSCs) in the treatment of Achilles tendinitis. Biomechanical indices of stiffness, stress, and maximum load-to-failure were detected by biomechanical test. mRNA and protein levels of miR-363, p-PI3K/AKT, tendon-related genes Collagen I, Scleraxis (Scx), and Tenascin C (TNC) were measured by qRT-PCR and western blot. The proliferation of hADSCs was accessed by MTT assay. Biomechanical indices of stiffness, stress, and maximum load-to-failure, and mRNA and protein levels of tendon-related genes could be improved by rhPDGF-BB or hADSCs alone, and could be further improved by rhPDGF-BB + hADSCs. rhPDGF-BB and hADSCs downregulated the expression of miR-363 and upregulated the levels of p-PI3K/Akt, and rhPDGF-BB + hADSCs further strengthened these effects. In addition, rhPDGF-BB promoted the proliferation of hADSCs in vitro and upregulated the expression of tendon-related genes. miR-363 mimic downregulated the levels of p-PI3K/Akt, miR-363 inhibitor upregulated the levels of p-PI3K/Akt, and miR-363 mimic and PI3K/Akt pathway inhibitor LY294002 reversed the positive effect of rhPDGF-BB on the proliferation of hADSCs, which suggested that rhPDGF-BB promoted the proliferation of hADSCs via miR-363/PI3K/Akt pathway. Biomechanical indices and tendon-related genes could be improved by rhPDGF-BB and hADSCs. Moreover, rhPDGF-BB promoted the proliferation of hADSCs via miR-363/PI3K/Akt pathway, indicating that rhPDGF-BB combined with ADSCs could treat Achilles tendinitis via miR-363/PI3K/Akt pathway.
为了探讨重组人血小板衍生生长因子-BB（rhPDGF-BB）和人脂肪来源干细胞（hADSCs）在治疗跟腱炎中的作用机制。通过生物力学测试检测了硬度、应力和最大破坏载荷等生物力学指标。通过 qRT-PCR 和 Western blot 检测了 miR-363、p-PI3K/AKT、肌腱相关基因胶原 I、Scleraxis（Scx）和 Tenascin C（TNC）的 mRNA 和蛋白水平。通过 MTT 测定法检测了 hADSCs 的增殖。rhPDGF-BB 或 hADSCs 单独处理可改善硬度、应力和最大破坏载荷等生物力学指标，以及肌腱相关基因的 mRNA 和蛋白水平，而 rhPDGF-BB+hADSCs 处理则可进一步改善这些指标。rhPDGF-BB 和 hADSCs 下调了 miR-363 的表达，上调了 p-PI3K/Akt 的水平，rhPDGF-BB+hADSCs 进一步增强了这些作用。此外，rhPDGF-BB 促进了 hADSCs 的体外增殖，并上调了肌腱相关基因的表达。miR-363 模拟物下调了 p-PI3K/Akt 的水平，miR-363 抑制剂上调了 p-PI3K/Akt 的水平，而 miR-363 模拟物和 PI3K/Akt 通路抑制剂 LY294002 逆转了 rhPDGF-BB 对 hADSCs 增殖的正向作用，这表明 rhPDGF-BB 通过 miR-363/PI3K/Akt 通路促进了 hADSCs 的增殖。rhPDGF-BB 和 hADSCs 可改善生物力学指标和肌腱相关基因。此外，rhPDGF-BB 通过 miR-363/PI3K/Akt 通路促进了 hADSCs 的增殖，表明 rhPDGF-BB 联合 ADSCs 通过 miR-363/PI3K/Akt 通路可治疗跟腱炎。
