实用医学杂志 ›› 2023, Vol. 39 ›› Issue (9): 1092-1097.doi: 10.3969/j.issn.1006⁃5725.2023.09.005

• 基础研究 • 上一篇    下一篇

瑞马唑仑通过调节肺泡巨噬细胞极化减轻脂多糖诱导的急性肺损伤 

王静1 高煜茹1 蔡钱伟3 朱委委1 黄潇1 孙大康2 王晓芝1 王涛1    

  1. 滨州医学院附属医院1 重症医学科,2 临床医学实验室(山东滨州 256603);3 山东大学齐鲁医院德州医院胸 外科(山东德州 253000)

  • 出版日期:2023-05-10 发布日期:2023-05-10
  • 通讯作者: 王涛 E⁃mail:wtsjr@163.com
  • 基金资助:
    山东省自然科学基金青年项目(编号:ZR2021QH105);中华国际医学交流基金(编号:Z⁃2018⁃35⁃2101) 

Remimazolam alleviates LPS ⁃ induced acute lung injury by regulating macrophage polarization 

WANG Jing* ,GAO Yuru,CAI Qianwei,ZHUA Weiwei,HUANG Xiao,SUN Dakang,WANG Xiaozhi,WANG Tao.   

  1. De⁃ partment of Critical Care Medicine,Binzhou Medical University Hospital,Binzhou 256603,China

  • Online:2023-05-10 Published:2023-05-10
  • Contact: Wang Tao E⁃mail:wtsjr@163.com

摘要:

目的 探讨瑞马唑仑对急性肺损伤(ALI)的作用及潜在分子机制。 方法 将 C57BL/6 雄性 小鼠随机分为正常对照组(Control 组)、模型组(LPS 组)、瑞马唑仑干预组(RM+LPS 组)和瑞马唑仑对照组 (RM 组),12 h 后处死小鼠取材,收集肺泡灌洗液(BALF)、血清和双侧肺组织。通过苏木素⁃伊红(HE)染色、肺组织的湿干质量比(W/D)、BCA 法检测 BALF 中总蛋白含量评价肺损伤情况;采用酶联免疫吸附实 验(ELISA)检测 BALF 和血清中肿瘤坏死因子⁃α(TNF⁃α)、白细胞介素(IL⁃6、IL⁃1β、IL⁃10)的含量;通过流 式细胞术检测 F4/80+ 标记的巨噬细胞中 M1 巨噬细胞阳性标志物 iNOS+ 和 M2 巨噬细胞标志物 CD206+ 的细胞比例;通过实时荧光定量 PCR 和蛋白质免疫印迹分析检测肺组织中 TNF⁃α、iNOS、CD206、Arg⁃1的mRNA和蛋白水平的表达量。结果 LPS 滴注 12 h 后小鼠出现明显肺损伤;BALF 和血清中 TNF⁃α、IL⁃6、 IL⁃1β 含量升高;流式细胞术检测 iNOS+ 细胞数量显著增加;肺组织中 TNF⁃α、iNOS mRNA 含量及蛋白水平 均显著增加。给予瑞马唑仑干预后,与 LPS 组相比,RM+LPS 组小鼠肺损伤明显改善;BALF 和血清中 TNF⁃ α、IL⁃6、IL⁃1β 含量显著降低,IL⁃10 含量显著增加;流式细胞术检测 iNOS+ 细胞数量显著降低,CD206+ 细胞 显著增加;肺组织中 TNF⁃α、iNOS mRNA 含量及蛋白水平均显著降低,CD206、Arg⁃1 mRNA 含量及蛋白水平均显著增加。结论 瑞马唑仑可通过抑制肺泡巨噬细胞向 M1 型极化,促进其向 M2 型极化,减轻炎症反应,缓解脂多糖诱导的ALI。

关键词:

脓毒症, 急性肺损伤, 巨噬细胞极化, 瑞马唑仑

Abstract:

Objective To investigate the effect and molecular mechanism of remimazolam on LPS⁃induced acute lung injury. Methods C57BL/6 mice were divided into normal control group,acute lung injury group,remimazolam pretreatment group and remimazolam control group,After 12 hours,the mice were sacrificed for BALF,serum and lung tissue collection. Lung injury was evaluated by HE,W/D ratio and total protein concentra⁃ tion in BALF. ELISA was used to detected the contents of TNF⁃α,IL⁃6,IL⁃1β,IL⁃10. The M1 macrophage marker iNOS + and the M2 macrophage marker CD206 + in F4/80 + macrophages were detected by flow cytometry and the mRNA and protein expressions of TNF ⁃ α,iNOS,CD206 and Arg1 were detected by real ⁃time quantitative PCR and Western blotting. Results Twelve hours after intratracheal injection of LPSA,the lung injury score,W/D ra⁃ tio and total protein concentration in BALF were significantly higher,the TNF⁃α,IL⁃6,IL⁃1β in BALF and serum was significantly increased,iNOS + macrophage was significantly increased,the mRNA and protein expressions of TNF⁃α and iNOS were significantly increased,as compared with the control group. After activated by remimazol⁃ am,the lung injury score,W/D ratio and total protein concentration in BALF were significantly decreased,the TNF⁃α,IL⁃6,IL⁃1β in BALF and serum was significantly decreased,IL⁃10 was significantly increased,iNOS + macrophage was significantly decreased,CD206 + macrophage was significantly increased,the mRNA and protein expressions of TNF⁃α and iNOS were significantly decreased,the mRNA and protein expressions of CD206 and Arg⁃ 1 were significantly increased,as compared with the LPS group. Conclusion Remimazolam alleviates LPS ⁃in⁃ duced acute lung injury by inhibiting macrophage polarization to M1 phenotype and promoting the macrophage po-larization to M2 phenotype.

Key words: sepsis, acute lung injury, macrophage polarization, remimazolam