南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (04): 525-530.doi: 10.12122/j.issn.1673-4254.2020.04.12

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丙泊酚抑制内毒素血症中巨噬细胞发生焦亡的分子机制

纪雪霞，郭远波，邱倩琪，王志鹏，王 研，吉锦泉，孙 强，蔡宇晶，周国斌   

  • 出版日期:2020-04-30 发布日期:2020-04-20
  • 基金资助:

Molecular mechanism underlying the inhibitory effect of propofol on lipopolysaccharideinduced pyroptosis of mouse bone marrow-derived macrophages

  

  • Online:2020-04-30 Published:2020-04-20

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摘要： 目的 探讨丙泊酚抑制巨噬细胞发生焦亡的分子机制。方法 提取小鼠骨髓来源巨噬细胞，分为对照组、脂多糖（LPS）+三磷酸腺苷（ATP）刺激组以及丙泊酚+LPS+ATP组。对照组不给予任何处理；LPS+ATP组先给予脂多糖(LPS) 1 μg/mL刺激4 h，再加三磷酸腺苷（ATP）4 mmol/L刺激1 h；丙泊酚+LPS+ATP组先同时给予50 μmmol/L丙泊酚+LPS 1 μg/mL刺激4 h，再加ATP刺激1 h。处理完毕则收集细胞培养上清液和细胞。分别通过CCK8、流式分析检测细胞活力，酶联免疫吸附测定法法检测细胞上清炎症因子IL-1β、IL-18含量；用免疫印迹检测细胞caspase-1蛋白表达及细胞膜Toll样受体-4（TLR4）表达；流式分析及免疫组化荧光检测细胞焦亡情况。结果 LPS+ATP可导致小鼠骨髓来源巨噬细胞（BMDM）活力显著降低（P<0.05）；炎症因子IL-1β、IL-18升高（P<0.05）；活化的caspase-1蛋白增加，细胞膜表面TLR-4表达升高（P<0.05）；予丙泊酚处理后，可改善LPS+ATP诱导的上述指标变化。结论 LPS+ATP可诱导BMDM发生焦亡，丙泊酚有效抑制这种细胞死亡，提示丙泊酚麻醉有益于临床脓毒症患者的手术，有效调节患者免疫状态。

Abstract: Objective To investigate the molecular mechanism underlying the inhibitory effect of propofol on pyroptosis of macrophages. Methods Macrophages derived from bone marrow were extracted and divided into three groups: control group, LPS+ATP group and propofol+LPS+ATP group. The control group was not given any treatment; LPS+ATP group was given LPS 1 μg/mL stimulation for 4 h, then ATP 4 mM stimulation for 1 h; Propofol+LPS+ATP group was given propofol+LPS 1 μg/mL stimulation for 4 h, then ATP stimulation for 1 h. After treatment, the supernatant and cells of cell culture were collected. the cell activity was detected by CCK8 and flow cytometry. The inflammatory cytokines IL-1βand IL-18 were detected by Elisa. Western blot was used to detect the expression of caspase-1 protein and TLR4 on cell membran Immunohistochemical fluorescence was used to detect apoptosis of cells. Results LPS+ATP significantly decreased the viability of the macrophages and increased the cellular production of IL-1β and IL-18, activation of caspase-1 protein and the expression of TLR-4 on the cell membrane (P<0.05). Treatment with propofol obviously reversed the changes induced by LPS+ATP. Conclusion LPS+ATP can induce pyroptosis of mouse bone marrow-derived macrophages, and propofol effectively inhibits such cell death, suggesting that propofol anesthesia is beneficial during operation and helps to regulate the immune function of in patients with sepsis.