Central Laboratory, Shanghai Sixth People's Hospital, Shanghai, 201306, China.
Department of Diagnostic and Interventional Radiology, Shanghai Sixth People's Hospital, Shanghai, 201306, China.
Cell Biochem Biophys. 2024 Dec;82(4):3695-3702. doi: 10.1007/s12013-024-01457-9. Epub 2024 Aug 3.
This study aimed to investigate the detrimental impact of cigarettes on lung cells and the potential effects of astragaloside IV on lung epithelial cell oxidative stress and pyroptosis. The research utilized cigarette smoke extract (CSE) to stimulate lung epithelial cells BEAS-2B, assessed cytotoxicity using the CCK-8 method, and measured changes in reactive oxygen species (ROS) and mitochondrial membrane potential with a probe method. Additionally, Seahorse XF24 was employed to analyze the impact of CSE on mitochondria in lung epithelial cells. Furthermore, LPS and cigarette combination-treated mice were created, alveolar damage was evaluated using HE staining, and changes in the key protein GSDMD of pyroptosis were detected using western blot (WB). The study also utilized the CCK-8 method to assess the potential toxic effects of astragaloside IV on lung epithelial cells, and the probe method to monitor changes in ROS and mitochondrial membrane potential. WB analysis was conducted to observe protein alterations in the TXNIP/NLRP3/GSDMD pathway. CSE concentration-dependently reduced cell activity, increased cellular ROS levels, and decreased mitochondrial membrane potential. CSE also decreases basal respiratory capacity, respiratory reserve capacity, and ATP production levels in cells. In LPS and cigarette combination-treated mice, cigarette smoke caused the alveolar septum to break and alveoli to enlarge, while increasing the expression of pyroptosis-related protein GSDMD. Astragaloside IV did not show significant cytotoxic effects within 48 h of treatment and could reduce CSE-induced ROS levels while increasing mitochondrial membrane potential. WB results indicated that astragaloside IV reduced the activation of the TXNIP/NLRP3/GSDMD signaling pathway in lung epithelial cells exposed to CSE. Our study demonstrates that CSE induces oxidative stress and impairs mitochondrial function in pulmonary epithelial cells, while astragaloside IV can potentially reverse these effects by inhibiting the TXNIP-NLRP3-GSDMD signaling pathway, thereby mitigating CSE-induced pulmonary disease and epithelial cell pyroptosis.
本研究旨在探讨香烟对肺细胞的有害影响,以及黄芪甲苷 IV 对肺上皮细胞氧化应激和细胞焦亡的潜在作用。研究采用香烟烟雾提取物(CSE)刺激肺上皮细胞 BEAS-2B,用 CCK-8 法评估细胞毒性,用探针法测量活性氧(ROS)和线粒体膜电位的变化。此外,还使用 Seahorse XF24 分析 CSE 对肺上皮细胞线粒体的影响。进一步构建 LPS 和香烟联合处理的小鼠模型,通过 HE 染色评估肺泡损伤,用 Western blot(WB)检测细胞焦亡关键蛋白 GSDMD 的变化。还采用 CCK-8 法评估黄芪甲苷 IV 对肺上皮细胞的潜在毒性作用,用探针法监测 ROS 和线粒体膜电位的变化。WB 分析用于观察 TXNIP/NLRP3/GSDMD 通路中蛋白的变化。CSE 浓度依赖性地降低细胞活性,增加细胞内 ROS 水平,降低线粒体膜电位。CSE 还降低细胞的基础呼吸能力、呼吸储备能力和 ATP 产生水平。在 LPS 和香烟联合处理的小鼠中,香烟烟雾导致肺泡间隔破裂,肺泡扩大,同时增加细胞焦亡相关蛋白 GSDMD 的表达。黄芪甲苷 IV 在治疗 48 小时内未表现出明显的细胞毒性作用,可降低 CSE 诱导的 ROS 水平,同时增加线粒体膜电位。WB 结果表明,黄芪甲苷 IV 可降低暴露于 CSE 的肺上皮细胞中 TXNIP/NLRP3/GSDMD 信号通路的激活。本研究表明,CSE 诱导肺上皮细胞氧化应激和线粒体功能障碍,而黄芪甲苷 IV 可能通过抑制 TXNIP-NLRP3-GSDMD 信号通路来逆转这些效应,从而减轻 CSE 诱导的肺部疾病和上皮细胞焦亡。