摘要： 目的 探讨低强度脉冲超声（LIPUS）对脂多糖（LPS）活化和未经活化的RAW264.7细胞迁徙和吞噬能力的影响。方法 LPS诱导RAW264.7巨噬细胞建立体外炎症活化模型；通过CCK-8法研究LIPUS对不同环境下RAW264.7细胞活力的影响，划痕实验观察LIPUS对RAW264.7巨噬细胞迁徙的影响，Transwell实验观察LPS作为趋化因素时RAW264.7细胞趋化迁徙能力的变化情况，共聚焦显微镜及流式细胞术分析细胞内FITC荧光强度以探究巨噬细胞吞噬pHrodo Green标记的大肠埃希菌生物颗粒的能力。结果 成功构建活化RAW264.7炎症模型，筛选LPS浓度为100 ng/mL；LIPUS抑制未活化的RAW264.7巨噬细胞向划痕区域内迁徙和未活化巨噬细胞的趋化迁徙；但在当前参数下LIPUS不影响巨噬细胞的细胞活力和吞噬能力。结论 LIPUS抑制经LPS诱导的RAW264.7巨噬细胞的迁徙，但不影响其吞噬作用。

低强度脉冲超声对脂多糖诱导炎性活化的RAW264.7巨噬细胞迁徙和吞噬的影响

郭雨萌, 王越, 刘笑涵, 吴琳

中国医科大学口腔医学院·附属口腔医院修复科, 辽宁省口腔疾病重点实验室, 沈阳 110002

收稿日期:2023-04-12出版日期:2024-01-30发布日期:2024-01-09
通讯作者:吴琳E-mail:wulin13@163.com
作者简介:郭雨萌(1995-),女,硕士研究生.
基金资助:国家自然科学基金（82201104）


关键词: 低强度脉冲超声, 脂多糖, 巨噬细胞, 迁徙, 吞噬
Abstract: Objective Tto evaluate the effect of low intensity pulsed ultrasound (LIPUS) on the migration and phagocytosis ability of lipopolysaccharide (LPS)-induced RAW264.7 macrophages and on macrophage behavior under inflammation. Methods An in vitro activated RAW264.7 macrophage model was developed using LPS. Cell viability was assessed using a CCK-8 assay to explore the effect of LIPUS on activated and inactivated macrophages. Wound healing assays were employed to measure the effect of LIPUS on macrophage migration, and the Transwell assay was employed when LPS was used as a chemoattractant. Phagocytosis ability was examined using confocal microscopy and flow cytometry by observing the FITC fluorescence signal of internalized pHrodo Green E. coli BioParticles Conjugate. Results An activated RAW264.7 macrophage model was successfully developed using 100 ng/mL LPS. LIPUS inhibited the migration of inactivated macrophages into scratch areas as well as guided cell migration. However, cell viability and phagocytosis remained unchanged. Conclusion LIPUS may inhibit RAW264.7 migration but not affect the phagocytosis ability of the macrophages.
Key words: low intensity pulsed ultrasound, lipopolysaccharide, macrophage, migration, phagocytosis
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