文章摘要
陈 绩,杨传熙,徐天华,孙 伟,孔祥清,盛燕辉.低强度脉冲式超声波对小鼠骨髓来源巨噬细胞极化和吞脂能力的影响[J].南京医科大学学报,2021,(4):489~495
低强度脉冲式超声波对小鼠骨髓来源巨噬细胞极化和吞脂能力的影响
LIPUS improves murine bone marrow⁃derived macrophage polarization and its phagocytosis of lipid droplet
投稿时间:2020-08-16  
DOI:10.7655/NYDXBNS20210403
中文关键词: 炎症  低强度脉冲式超声波  巨噬细胞  M1表型  吞脂能力  NF⁃κB/MAPK通路
英文关键词: inflammation  LIPUS  macrophage  M1 phenotype  phagocytosis of lipid droplet  NF⁃κB/MAPKpathway
基金项目:国家重大科研仪器研制项目(81627802)
作者单位
陈 绩 南京医科大学第一附属医院心内科江苏 南京 210029 
杨传熙 东南大学医学院江苏 南京 211189 
徐天华 南京医科大学第一附属医院心内科江苏 南京 210029 
孙 伟 南京医科大学第一附属医院心内科江苏 南京 210029 
孔祥清 南京医科大学第一附属医院心内科江苏 南京 210029 
盛燕辉 南京医科大学第一附属医院心内科江苏 南京 210029 
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中文摘要:
      目的:探究低强度脉冲式超声波(low?intensity pulsed ultrasound,LIPUS)对巨噬细胞极化与吞脂能力的潜在影响及其相关分子机制。方法:采用胫骨骨髓灌注法提取小鼠骨髓来源的巨噬细胞,分别用不同强度(4.825、19.300、43.425 mW/cm2)的LIPUS对巨噬细胞进行辐照处理,再给予脂多糖(lipopolysaccharide,LPS)诱导巨噬细胞向M1型极化;采用流式细胞术检测巨噬细胞凋亡水平;CCK?8法检测巨噬细胞活性;热电偶测温技术检测LIPUS热效应;实时荧光定量PCR(qPCR)分析M1型巨噬细胞标志基因的mRNA水平;Western blot检测NF?κB/MAPK 通路相关蛋白表达;采用荧光染料DiI标记的氧化低密度脂蛋白与巨噬细胞共同孵育6 h,荧光显微镜观察并计算巨噬细胞的脂滴吞噬效率。结果:流式细胞术和CCK?8结果表明,不同强度LIPUS辐照处理均未对巨噬细胞的凋亡和细胞活性产生影响;热电偶测温技术表明,温度变化稳定,未产生明显热效应。qPCR结果显示,与对照组相比,LPS能显著上调巨噬细胞M1表型基因的mRNA水平,而LIPUS辐照处理可下调LPS诱导的M1表型基因mRNA水平,差异具有统计学意义;脂滴吞噬染色结果显示,LPS能够增强巨噬细胞的吞脂作用,而LIPUS辐照处理能减弱LPS对巨噬细胞吞脂能力的促进作用,差异具有统计学意义;Western blot结果显示,LIPUS能够下调LPS诱导的NF?κB/MAPK通路蛋白水平,差异具有统计学意义。结论:LIPUS可能通过抑制LPS活化的NF?κB/MAPK通路阻止巨噬细胞向M1型极化以及减弱巨噬细胞的吞脂能力。
英文摘要:
      Objective:We aimed to investigate the potential effects of low?intensity pulsed ultrasound(LIPUS)on the polarization and lipid droplet phagocytosis of macrophage,and further explore the underlying molecular mechanism. Methods:Murine tibial bone marrow was perfused to extract murine bone marrow?derived macrophage. Different intensities(4.825,19.300,43.425 mW/cm2)of LIPUS treatment were performed in macrophage. Then LPS was used to induce polarization of macrophage to the M1 phenotype. After that,we used flow cytometry and CCK?8 kit to determine the levels of apoptosis and viability of macrophage in different groups,respectively. Besides,the heating effects of LIPUS irradiation were measured by thermocouples. The mRNA expression of inflammatory?related genes was validated by q?PCR analysis. Western blot was conducted to explore the protein levels of NF?κB/MAPK pathway. Then,the phagocytosis of DiI?labeled OX?LDL was observed under inverted fluorescence microscope after 6 h co?culture with macrophage. Results:First,no significant difference of cell apoptosis in LIPUS group was shown in flow cytometry compared with that in the control group. The viability of macrophage examined in CCK?8 did not behave obvious difference. Besides,the results of temperature test demonstrated that the ultrasound conditions we set hardly changed the temperature of culture medium during LIPUS procedures. Then,results of q?PCR showed that mRNA levels of M1 phenotype genes were remarkedly up?regulated in LPS?induced macrophage compared with those in the control group,which was attenuated by LIPUS treatment(P < 0.05). Moreover,LIPUS treatment down regulated LPS?induced phagocytosis of DiI?labeled OX?LDL(P < 0.05). Results of western blot showed that LIPUS inhibited LPS?induced activation of NF?κB/MAPK pathways(P < 0.05). Conclusion:Overall,our results demonstrated that LIPUS could reduce LPS?induced polarization of macrophage to the M1 phenotype,and phagocytosis of lipid droplet through inactivating NF?κB/MAPK pathway.
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