【摘要】 目的 构建小鼠胚胎成骨细胞前体细胞（MC3T3-E1）衰老模型，通过检测细胞衰老和成骨分化相关指标，验证血小板衍生生长因子-BB（PDGF-BB）对MC3T3-E1细胞衰老以及成骨分化的作用，并探索其机制。方法 使用H₂O₂诱导MC3T3-E1细胞衰老，并进行PDGF-BB干预。检测细胞衰老标志物包括细胞增殖、细胞衰老β-半乳糖苷酶（SA-β-gal）染色阳性细胞数、活性氧（ROS）生成、周期蛋白依赖激酶抑制剂2A （P16）和周期蛋白依赖激酶抑制剂1A（P21）以及炎性相关因子白介素-6（IL-6）、白介素-8（IL-8）的mRNA表达水平和成骨分化相关指标。使用抑制剂MK-2206探索AKT信号通路。结果 与Control组相比，H₂O₂组细胞增殖减弱，SA-β-gal染色阳性细胞数、ROS生成增多，P16、P21、IL-6和IL-8的基因表达上调（P<0.05）以及成骨分化能力受损（P<0.05）;H₂O₂+PDGF-BB组细胞的增殖能力、成骨分化能力高于H₂O₂组（P<0.05）, SA-β-gal染色阳性细胞数、ROS生成、P16、P21、IL-6和IL-8基因表达水平低于H₂O₂组（P<0.05）;H₂O₂+PDGF-BB+MK-2206组细胞的增殖能力、成骨分化能力低于H₂O₂+PDGF-BB组（P<0.05）,SA-β-gal染色阳性细胞数、ROS生成、P16、IL-6和IL-8基因表达水平高于H₂O₂+PDGF-BB组（P<0.05）。结论 PDGF-BB对抗小鼠MC3T3-E1细胞衰老并促进衰老细胞的成骨分化，该作用的发挥可能与AKT信号通路被再次激活有关。更多还原

【Abstract】 Objective To establish an in vitro senescence model in mouse embryonic osteoblast precursor cells（MC3T3-E1） and to investigate the effects of platelet-derived growth factor-BB（PDGF-BB） on senescence and osteogenic differentiation of MC3T3-E1 cells, and the potential mechanisms by detecting cell senescence and osteogenic differentiation related indicators.Methods MC3T3-E1 cells were induced with H₂O₂ to create the in vitro senescence model, followed by PDGF-BB intervention. Cell senescence markers were assessed, including cell proliferation, senescence-associated β-galactosidase（SA-β-gal） staining-positive cells, reactive oxygen species（ROS） generation, mRNA expressions of cyclin-dependent kinase inhibitors 2A（P16） and 1A（P21）, as well as inflammatory factors interleukin-6（IL-6） and interleukin-8（IL-8）, and osteogenic differentiation markers. The protein kinase B（AKT） signaling pathway was explored using the inhibitor MK-2206.Results Compared to the Control group, the H₂O₂ group exhibited decreased cell proliferation, increased SA-β-gal-positive cells, elevated ROS generation, and up-regulated P16, P21, IL-6, and IL-8（P<0.05）, as well as impaired osteogenic differentiation ability（P<0.05）. The H₂O₂+PDGF-BB group showed significantly higher cell proliferation and osteogenic differentiation capacity than the H₂O₂ group（P<0.05）, but less number of SA-β-gal-positive cells, and lower ROS production, and expression levels of P16, P21, IL-6, and IL-8（P<0.05）. In contrast, the H₂O₂+PDGF-BB+MK-2206 group demonstrated significantly lower cell proliferation and osteogenic differentiation ability compared to the H₂O₂+PDGF-BB group（P<0.05）, but higher SA-β-gal-positive cells, ROS generation, and expression levels of P16, IL-6, and IL-8（P<0.05）.Conclusion PDGF-BB can counteract cell senescence in murine MC3T3-E1 cells and promote osteogenic differentiation of senescent cells by reactivating the AKT signaling pathway.更多还原