[Molecular mechanism underlying the inhibitory effect of propofol on lipopolysaccharide-induced pyroptosis of mouse bone marrow-derived macrophages]

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. 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。

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.

To investigate the molecular mechanism underlying the inhibitory effect of propofol on pyroptosis of macrophages.

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. Conclusions: 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. 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。