异氟醚通过Nrf2/HO-1/ROS途径抑制缺氧引起肺动脉平滑肌细胞焦亡  

Isoflurane Inhibits Pyroptosis of Human Pulmonary Artery Smooth Muscle Cells Through the Nrf2/HO-1/ROS Pathway

作　　者：刘文志 郭立平[1] 张倩璐[1] LIU Wen-Zhi;GUO Li-Ping;ZHANG Qian-Lu(Department of Anesthesiology,The First Affiliated Hospital of University of South China,Hengyang 421001,Hunan,China)

机构地区：[1]南华大学第一附属医院麻醉科,湖南衡阳421001

出　　处：《中国生物化学与分子生物学报》2022年第8期1087-1097,共11页Chinese Journal of Biochemistry and Molecular Biology

基　　金：衡阳市科学技术项目(No.2016KJ41)资助。

摘　　要：研究发现,异氟醚吸入麻醉可明显减轻由缺血-再灌注引起的肺动脉高压(PAH),提示其对肺循环功能有一定保护效应。肺动脉平滑肌细胞(PASMC)是肺动脉血管重塑和PAH发生的主要参与者,其结构改变和功能异常均可显著影响肺动脉高压病情进展。本研究探讨异氟醚对缺氧诱导的PASMC焦亡的影响及其调控机制,旨在为肺动脉高压治疗提供潜在分子靶点。PASMC于37℃、5%CO_(2)、3%O_(2)条件下静置培养24 h建立缺氧模型。RT-PCR和Western印迹等结果显示,缺氧致使PASMC内紅系衍生的核转录因子2(Nrf2)核转位减少,血红素加氧酶-1(HO-1)表达水平下调,而焦亡相关蛋白质,包括NOD样受体蛋白3(NLRP3)、胱天蛋白酶1(caspase-1)、凋亡相关斑点样蛋白(ASC)及消皮素D(GSDMD)等表达上调,活性氧(ROS)生成、胱天蛋白酶1活性和乳酸脱氢酶(LDH)释放水平升高,Hoechst/PI染色显示,焦亡孔洞增加。ELISA结果表明,IL-1β、IL-6、IL-18和TNF-α分泌增加(P<0.05)。异氟醚处理可显著激活Nrf2/HO-1通路,缓解缺氧诱导的PASMC焦亡,并且ROS阻断剂NAC预处理肺动脉平滑肌细胞,可使异氟醚的抗焦亡效应更为明显。另外,Nrf2特异性siRNA(Nrf2 siRNA)、或HO-1抑制剂锌原卟啉(Znpp)处理肺动脉平滑肌细胞,异氟醚引起的ROS生成显著升高(P<0.05),且Nrf2 siRNA转染显著抵消了异氟醚的抗焦亡效应。综上,异氟醚可通过Nrf2/HO-1/ROS途径抑制缺氧引起的肺动脉平滑肌细胞焦亡。Previous studies have demonstrated that isoflurane inhale anesthesia can effectively attenuate the ischemia-reperfusion-induced pulmonary hypertension(PAH),indicating a protective effect of isoflurane on pulmonary circulation.Pulmonary artery smooth muscle cells(PASMCs)play an important role in pulmonary vascular remodeling and PAH.The abnormality of PASMC structure and function may greatly contribute to the development of PAH.This study aims to explore the effects of isoflurane on hypoxia-induced PASMC pyroptosis and the underlying mechanisms,and to find potential therapeutic target for the treatment of PAH.PASMCs were cultured at 37℃,5%CO_(2) and 3%O_(2) for 24 h to establish a hypoxia model.RT-PCR and Western blotting showed that hypoxia treatment significantly decreased Nrf2 translocation and HO-1 expression,while the levels of pyroptosis-related proteins(NLRP3,caspase-1,ASC,GSDMD),caspase-1 activity,ROS production,LDH release were up-regulated.Plus,the Hoechst/PI staining and ELISA assay showed that the pore formation and secretion of pro-inflammatory cytokines(IL-1β,IL-6,IL-18,TNF-α)were remarkably increased(P<0.05).Treatment with isoflurane significantly ameliorated hypoxia-induced pyroptosis,which could be amplified by pre-incubation with the ROS inhibitor,NAC.Furthermore,transfection of PASMCs with Nrf2 siRNA or pre-treatment with the HO-1 inhibitor,Znpp,notably elevated isoflurane-induced ROS production.The transfection of Nrf2 siRNA also compromised the effects of isoflurane on PASMC pyroptosis(P<0.05).Taken together,we deduce the conclusion that isoflurane inhibits hypoxia-induced PASMC pyroptosis through the Nrf2/HO-1/ROS pathway.

关 键 词：异氟醚 核因子E2相关因子2 活性氧 肺动脉平滑肌细胞 焦亡 

分 类 号：R614[医药卫生—麻醉学] R365[医药卫生—外科学]