摘要
目的 探究罗哌卡因对缺氧/复氧(H/R)诱导A549细胞氧化应激、炎症反应和铁死亡的影响。方法 取人Ⅱ型肺泡上皮细胞A549分为对照组(Control组,未予任何处理的A549细胞)、模型组(H/R组)、罗哌卡因组(予H/R联合不同浓度罗哌卡因处理)。采用MTT法检测不同浓度罗哌卡因对A549细胞活性的影响,采用TUNEL分析不同浓度罗哌卡因对A549细胞凋亡的影响,并使用相关试剂盒测定不同浓度罗哌卡因对A549细胞氧化应激、炎症反应和铁死亡相关指标的影响,采用Western印迹法检测不同浓度罗哌卡因对相关蛋白表达的影响。结果 与Control组比较,H/R组A549细胞活力降低,细胞凋亡率显著升高,丙二醛(MDA)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)和活性氧(ROS)、Fe^(2+)水平升高,超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)水平降低,FTH1和GPX4表达水平显著下调,Tf、Keap1、Nrf2和HO-1表达显著上调(P<0.05,P<0.01)。与H/R组比较,5.00、10.00、20.00μmol/L罗哌卡因组A549细胞活力升高,细胞凋亡率显著降低,SOD、GSH-Px、FTH1、GPX4表达降低和MDA、IL-6、IL-8、TNF-α、ROS、Fe^(2+)及Tf表达升高被显著逆转,Keap1蛋白表达降低,Nrf2和HO-1蛋白表达升高(P<0.05,P<0.01)。结论 罗哌卡因在H/R诱导的肺上皮细胞损伤中发挥保护作用,且可能与Keap1-Nrf2/HO-1信号通路活化有关。
Objective To investigate the effects of Ropivacaine on oxidative stress,inflammatory response and ferroptosis in A549 cells induced by hypoxia/reoxygenation(H/R).Methods Human type II alveolar epithelial cells A549 were divided into Control group(A549 cells did not receive any treatment),model group(H/R group) and Ropivacaine group(H/R combined with different concentrations of Ropivacaine).MTT assay was used to detect the effects of different concentrations of Ropivacaine on the activity of A549 cells,and TUNEL was used to analyze the effects of different concentrations of Ropivacaine on the apoptosis of A549 cells.The effects of different concentrations of Ropivacaine on oxidative stress,inflammatory response and ferroptosis-related indicators in A549 cells were measured using relevant kits,and the effect of Ropivacaine on the expression of related proteins was detected by Western blotting.Results Compared with Control group,A549 cells in H/R group had decreased viability,significantly increased apoptosis rate,and increased levels of malondialdehyde(MDA),interleukin-6(IL-6),interleukin-8(IL-8),tumor necrosis factor-α(TNF-α),reactive oxygen species(ROS) and Fe^(2+).The levels of superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) were decreased,the expressions of FTH1 and GPX4 were significantly down-regulated,and the expressions of Tf,Keap1,Nrf2 and HO-1 were significantly up-regulated(P<0.05,P<0.01).Compared with H/R group,A549 cells in 5.00,10.00 and 20.00 μmol/L Ropivainine groups had increased viability,significantly decreased apoptosis rate,decreased SOD,GSH-Px,FTH1 and GPX4 expression,significant reversion of increased MDA,IL-6,IL-8,TNF-α,ROS,Fe^(2+) and Tf expression,decreased Keap1 protein expression,and increased Nrf2 and HO-1 protein expression(P<0.05,P<0.01).Conclusion Ropivacaine plays a protective role in H/R-induced alveolar epithelial cell injury,which may be associated with the activation of Keap1-Nrf2/HO-1 signaling pathway.
作者
尹亚岚
钟惠
鲍蕾
赵春武
李金凤
周璐
YIN Yalan;ZHONG Hui;BAO Lei;ZHAO Chunwu;LI Jinfeng;ZHOU Lu(Department of Anesthesiology,the Seventh People's Hospital of Chengdu City,Chengdu 610000,China)
出处
《临床误诊误治》
CAS
2023年第7期44-49,共6页
Clinical Misdiagnosis & Mistherapy
基金
四川省医学青年创新科研课题计划(Q1906315)。
关键词
罗哌卡因
肺上皮细胞
氧化应激
炎症反应
铁死亡
丙二醛
白细胞介素-8
超氧化物歧化酶
Ropivacaine
Pulmonary epithelial cell
Oxidative stress
Inflammatory response
Ferroptosis
Malondialdehyde
Interleukin-8
Superoxide dismutase
