力学刺激对巨噬细胞极性的影响
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上海市骨科内植物重点实验室开放课题(KFKT2019002)


Effects of Mechanical Stimulation on Polarity of Macrophages
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    摘要:

    目的 探讨力学刺激对巨噬细胞极性的影响。方法 使用不同幅度、时间张应变刺激RAW264.7细胞,CCK-8试剂检测细胞活性以明确使用的力学刺激参数。将RAW264.7细胞诱导为M1型巨噬细胞,并施加10%幅度、2 Hz张应变。使用CCK-8、流式细胞仪检测张应变对细胞活性、凋亡的影响。运用实时荧光定量聚合酶链反应仪(quantitative real-time polymerase chain reaction, qRT-PCR)检测张应变对M1型巨噬细胞炎症相关基因表达量的影响。结果 在刺激3 h后,15%和20%幅度、2 Hz张应变会显著抑制细胞活性(P<0.05),而10%幅度、2 Hz张应变不会对RAW264.7活性产生抑制作用(P>0.05)。10%幅度、2 Hz张应变对M1型巨噬细胞凋亡、活性无显著影响(P>0.05)。10%幅度、2 Hz张应变可以显著抑制M1型巨噬细胞炎症相关因子白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)的表达(P<0.05)。结论 10%幅度、2 Hz力学刺激可抑制M1型巨噬细胞,促使其向M2型转化。力学刺激可能成为炎症相关疾病的一种治疗手段。

    Abstract:

    Objective To explore the effect of mechanical stimulation on polarity of macrophages. Methods RAW264.7 cells were stimulated with tensile stretch at various amplitude and time, then cell viability was assessed with cell count kit-8 (CCK-8) for determining the stimulation parameters. RAW264.7 cells were induced to M1 type, then tensile stretch at 10% amplitude and 2 Hz was applied to M1 cells. CCK-8 and flow cytometry were used to detect the effects of tensile stretch on cell activity and apoptosis. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the effect of tensile stretch on M1 type macrophage related gene expression. Results After stimulation for 3 hours, tensile stretch at 15% or 20% amplitude and 2 Hz significantly inhibited cell viability (P<0.05), while tensile stretch at 10% amplitude and 2 Hz did not inhibit the viability of RAW264.7 cells (P>0.05). Tensile stretch at 10% amplitude and 2 Hz neither inhibited viability nor cause apoptosis of M1 type macrophages. The expression of inflammation-related genes including interleukin-1β(IL-1β) and tumor necrosis factor-α (TNF-α) of M1 type macrophages was significantly down-regulated with tensile stretch at 10% amplitude and 2 Hz (P<0.05). Conclusions Mechanical stimulation at 10% amplitude and 2 Hz can inhibit M1 type macrophages and promote the polarization from M1 to M2. Mechanical stimulation may become a method for treating inflammation-related diseases.

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池广浩,李般若,吴伟,郝敏,马建,邱龙顺.力学刺激对巨噬细胞极性的影响[J].医用生物力学,2020,35(6):739-743

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  • 收稿日期:2019-09-05
  • 最后修改日期:2020-02-01
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  • 在线发布日期: 2020-12-24
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