摘要： 目的 探讨丙泊酚作用核苷酸结合寡聚化结构域样受体蛋白3 （NLRP3）/凋亡相关斑点样蛋白（ASC）/半胱氨酸天冬氨酸蛋白酶-1 （caspase-1）通路对A549细胞焦亡的影响。方法 超低吸附培养法建立肺癌A549细胞三维培养模型，建立模型后采用不同浓度丙泊酚作用肺癌A549细胞，CCK-8法检测丙泊酚对A549细胞增殖的影响；酶联免疫吸附法检测肺癌A549细胞上清炎性细胞因子白细胞介素（IL）-18、IL-1β、IL-6表达水平变化；蛋白免疫印迹法检测各组焦亡相关蛋白NLRP3、ASC、caspase-1、焦孔素蛋白N端（GSDMD-N）、IL-1β表达水平。结果 三维培养下，A549细胞间有聚拢黏附趋势，培养12 h即可清晰观察到细胞球。与空白对照组相比，随着丙泊酚浓度增加，肺癌A549细胞存活率依次降低（P < 0.05）；肺癌A549细胞上清炎性细胞因子IL-18、IL-1β、IL-6、焦亡相关蛋白NLRP3、ASC、caspase-1、GSDMD-N、IL-1β表达水平依次增加（P < 0.05）。结论 超低吸附培养法成功建立肺癌A549细胞三维培养模型，丙泊酚通过激活NLRP3/ASC/caspase-1通路促进细胞焦亡，进而抑制肺癌A549细胞增殖。

丙泊酚作用NLRP3/ASC/caspase-1通路对肺癌A549细胞焦亡的影响

关冀弛, 杨金雨, 刘丹, 李磊, 李东华, 汪海峰

沈阳化工大学制药与生物工程学院, 沈阳 110142

收稿日期:2023-03-02出版日期:2024-02-28发布日期:2024-01-12
通讯作者:汪海峰E-mail:38whf@163.com
作者简介:关冀弛(1998-),男,硕士研究生.
基金资助:辽宁省教育厅科学研究经费项目青年科技人才“育苗”项目（LQ2020021）


关键词: 丙泊酚, 超低吸附培养法, 肿瘤细胞三维培养模型, 肺癌, 细胞焦亡
Abstract: Objective To investigate the effect of propofol on pyroptosis and A549 cells via the NLRP3/ASC/caspase-1 pathway. Methods Establish a three-dimensional culture model of A549 tumor cells using ultra-low attachment plates, A549 cells were cultured using ultra-low adsorption culture plates to establish a three-dimensional culture model. The CCK-8 method was used to detect the effect of propofol on A549 cell proliferation;the inflammatory factors interleukin (IL) -18, IL-1β, and IL-6 were detected in the A549 lung cancer cell supernatants using enzyme-linked immunosorbent assays;western blotting was used to detect the expression levels of pyrolysis-associated proteins NLRP3, ASC, caspase-1, GSDMD-N, and IL-1β in A549 lung cancer cells in each group. Results Compared to the blank control group, the survival rate of A549 cells in low, medium, and high concentrations of propofol in each group decreased in turn (P < 0.05);the levels of inflammatory factors IL-18, IL-1β, and IL-6 in the A549 cell supernatant, and scorch related protein NLRP3, ASC, caspase-1, GSDMD-N, and IL-1β increased with the increased propofol concentrations (P < 0.05). Conclusion The three-dimensional culture model of lung cancer A549 cells was successfully established using the ultra-low adsorption culture method. Propofol can promote cell apoptosis and inhibit the pyroptosis of A549 lung cancer cells via activating the NLRP3/ASC/caspase-1 pathway.
Key words: propofol, ultra-low attachment plates, three-dimensional culture model of tumor cells, lung cancer, pyroptosis
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