摘要
目的 研究囊性纤维化跨膜转导调节因子(CFTR)氯离子通道在Ⅱ型肺泡上皮细胞(AECⅡ)高氧损伤中的作用。方法 AECⅡ于密封舱中通入95%O_(2)+5%CO_(2),持续24h或48h,构建高氧损伤AECⅡ模型。将实验组AECⅡ随机分为高氧24h组、高氧24h+激动剂(Gen)组、高氧24h+抑制剂(AA)、高氧48h组、高氧48h+Gen组和高氧48h+AA组;对照组通入空气,未干预。以膜片钳系统记录CFTR氯离子电流强度。细胞外液中分别加入GEN50μM或AA10μM,记录GEN激活或AA抑制后氯离子电流强度。采用Clampfit 10.6行数据分析,应用R4.4.1统计软件对不同组别行Spearman相关性分析。结果 1.高氧24h、48h明显抑制AECⅡ氯离子电流(P<0.01),以24h较明显;2. Gen明显增加高氧24h、48hAECⅡ氯离子电流;AA明显抑制氯离子电流(P<0.01),均以48h较明显。结论 高氧可能通过影响AECⅡCFTR氯离子外流而发挥作用。
Objective To investigate the role of cystic fibrosis transmembrane conductance regulator(CFTR)chloride channel in high oxygen-induced injury of alveolar epithelial cells type II(AECⅡ).Methods AECsⅡwere Placed in a sealed chamber containing 95%O2 and 5%CO2,and incubated in a culture incubator for 24 hours or 48 hours to establish a model of high oxygen damage in AECsⅡ.In the experimental group,AECsⅡwere randomly divided into the following groups:the hyperoxia group for 24 hours,the hyperoxia group for 24 hours plus the stimulant(Gen),the hyperoxia group for 24 hours plus the inhibitor(AA),the hyperoxia group for 48 hours,the hyperoxia group for 48 hours plus Gen,and the hyperoxia groupfor 48 hours plus AA.The control group AECsⅡwere cultured in a medium with air and without intervention.The whole-cell Patch-clamp system was used to record the chloride current strength of CFTR which was recorded after Gen at 50μM or AA at 10μM was added in the extracellular fluid.Data was analyzed using Clampfit 10.6,where the current changes under different stimulation voltages were calculated,and the results were presented in the form of a voltage-current trend line.Spearman correlation analysis was conducted on the different groups using R 4.4.1 statistical software.Results 1.ComPared with the control group,high oxygen exposure for 24 hours and 48 hours significantly inhibited the chloride current of AECⅡ,showing a decrease in the negative value(P<0.01),with the 24-hour group showing a more significant inhibition.2.Compared with the simple hyPeroxia group,Gen significantly increased the chloride current of AECⅡin high oxygen for 24 hours and 48 hours,showing an increase in the negative value(P<0.01),with the 48-hour group being more obvious(P<0.01);while AA significantly inhibited the chloride current,showing a decrease in the negative value(P<0.01),with the 48-hour group being more obvious(P<0.01).Conclusion Hyperoxia may exert its effects by affecting the chloride current outflow of CFTR in AECⅡ.
作者
葛丹丹
刘登礼
GE Dan-dan;LIU Deng-li(Department of Pediatrics,Pediatric Key Laboratory of Xiamen,Institute of Pediatrics,School of Medicine,the First Affiliated Hospital of Xiamen University,Xiamen 361003,Fujian Province,China)
出处
《罕少疾病杂志》
2025年第1期54-56,共3页
Journal of Rare and Uncommon Diseases
基金
福建省自然科学基金(2020J011223)。
