固本葆骨方对糖皮质激素相关性骨微血管内皮细胞损伤的保护作用及机制研究
Protective effects of Gumibao recipe on glucocorticoid-included bone microcirculatory endothelial cell injury and the underlying mechanism.
机构信息
Orthopedics and Traumatology Department, Suzhou Hospital of Traditional Chinese Medicine, Suzhou 215000, China.
Orthopedics and Traumatology Department, Suzhou Hospital of Traditional Chinese Medicine, Suzhou 215000, China.
出版信息
Int Immunopharmacol. 2024 Dec 5;142(Pt A):112989. doi: 10.1016/j.intimp.2024.112989. Epub 2024 Aug 31.
OBJECTIVE
To investigate the protective effects of Gumibao recipe on glucocorticoid-included bone microcirculatory endothelial cell (BMEC) injury, and elucidate the possible underlying mechanism.
METHODS
BMECs were treated with different concentrations of hydrocortisone at different time points, and the viability as well as migration of BMECs were evaluated; furthermore, the release of LDH, levels of VEGF, PAI-1, t-PA, and the content of NO by BMECs have been evaluated by commercially available kits; moreover, the expressions of eNOS, p-PI3K, p-Akt and p-mTOR in BMECs were examined by WB methods. Next, hydrocortisone treated BMECs were co-treated with Gumibao recipe, and the viability, migration and autophagy of BMECs were evaluated.
RESULTS
0.2 mg/ml and 0.3 mg/ml hydrocortisone significantly decreased viability and migration ability of BMECs, and also impeded the endothelial function of BMECs by decreasing the levels of VEGF, t-PA, the content of NO, and increasing the level of PAI-1. Gumibao medicated serum markedly increased the viability and migration of BMECs, and also increased the levels of VEGF, t-PA, the content of NO, meanwhile decreased the level of PAI-1 in 0.3 mg/ml hydrocortisone treated BMECs; moreover, glucocorticoids inhibited the autophagy of BMECs, and Gumibao recipe significantly increased the autophagy of BMECs; meanwhile, autophagy inhibitor 3-MA partially blocked the protective effects of Gumibao recipe. Finally, gumibao recipe partially abrogated the inhibitory effects of hydrocortisone on the activation of PI3K/Akt/mTOR singling, and these effects were further counteracted by PI3K and mTOR inhibitor NVP-BEZ235.
CONCLUSIONS
We reported for the first time the protective effects of Gumibao recipe on glucocorticoid-included BMECs injury, and the possible underlying mechanism may be regulating the autophagy of BMECs via PI3K/AKT/mTOR signaling pathway.
目的
探讨龟芪骨肽方对糖皮质激素诱导的骨髓微血管内皮细胞（BMEC）损伤的保护作用，并阐明其可能的作用机制。
方法
采用不同浓度的氢化可的松处理 BMECs 不同时间，评估 BMECs 的活力和迁移能力；同时，采用商业试剂盒评估 BMECs 乳酸脱氢酶（LDH）的释放、血管内皮生长因子（VEGF）、纤溶酶原激活物抑制剂-1（PAI-1）、组织型纤溶酶原激活物（t-PA）和一氧化氮（NO）的水平；此外，采用 Western blot 方法检测 BMECs 中内皮型一氧化氮合酶（eNOS）、磷酸化磷脂酰肌醇 3 激酶（p-PI3K）、磷酸化蛋白激酶 B（p-Akt）和磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）的表达。接下来，用龟芪骨肽方共同处理氢化可的松处理的 BMECs，评估 BMECs 的活力、迁移和自噬。
结果
0.2mg/ml 和 0.3mg/ml 的氢化可的松显著降低了 BMECs 的活力和迁移能力，并通过降低 VEGF、t-PA、NO 的水平和增加 PAI-1 的水平，损害了 BMECs 的内皮功能。龟芪骨肽方明显增加了 0.3mg/ml 氢化可的松处理的 BMECs 的活力和迁移，并增加了 VEGF、t-PA、NO 的水平，同时降低了 PAI-1 的水平；此外，糖皮质激素抑制了 BMECs 的自噬，而龟芪骨肽方显著增加了 BMECs 的自噬；同时，自噬抑制剂 3-MA 部分阻断了龟芪骨肽方的保护作用。最后，龟芪骨肽方部分阻断了氢化可的松对 PI3K/Akt/mTOR 信号通路的抑制作用，而这种作用被 PI3K 和 mTOR 抑制剂 NVP-BEZ235 进一步抵消。
结论
我们首次报道了龟芪骨肽方对糖皮质激素诱导的 BMECs 损伤的保护作用，其可能的作用机制可能是通过 PI3K/AKT/mTOR 信号通路调节 BMECs 的自噬。
Zotero 插件