氢气对屋尘螨诱导的鼻黏膜上皮细胞氧化应激损伤及炎症反应的影响  

Effects of hydrogen on oxidative stress damage and inflammatory response of nasal mucosal epithelial cells induced by house dust mites

作　　者：王雨洁 李茜 昂扬 龙云 张明燕 张婷 潘晗 陈伟[1] WANG Yujie;LI Qian;ANG Yang;LONG Yun;ZHANG Mingyan;ZHANG Ting;PAN Han;CHEN Wei(Department of Otorhinolaryngology Head and Neck Surgery,Jinling Clinical Medical College,Nanjing University of Traditional Chinese Medicine/General Hospital of Eastern Theater Command,PLA,Nanjing 210002,Jiangsu,China;Department of Anesthesiology and Pain,the Affiliated Jiangning Hospital of Nanjing Medical University,Nanjing 211100,Jiangsu,China;Department of Anesthesiology,Zhongda Hospital Southeast University,Nanjing 210009,Jiangsu,China;Department of Otorhinolaryngology Head and Neck Surgery,General Hospital of East-ern Theater Command,PLA,Nanjing 210002,Jiangsu,China)

机构地区：[1]南京中医药大学金陵临床医学院(东部战区总医院)耳鼻咽喉-头颈外科,南京210002 [2]南京医科大学附属江宁医院麻醉疼痛科,南京211100 [3]东南大学附属中大医院麻醉科,南京210009 [4]东部战区总医院耳鼻咽喉-头颈外科,南京210002

出　　处：《医学研究与战创伤救治》2025年第3期238-244,共7页Journal of Medical Research & Combat Trauma Care

基　　金：东部战区总医院院管课题(2023JCYJYB093,22LCZLXJS35)。

摘　　要：目的探讨氢气(H_(2))对屋尘螨提取物(HDM)诱导的人鼻黏膜上皮细胞(HNEPC)氧化应激的保护作用并推测其可能的机制。方法体外培养HNEPC细胞随机分为4组:NC组(不做任何处理)、HDM组(10μg/m L HDM构建AR体外模型)、HDM+H_(2)组、H_(2)组。采用CCK8法检测细胞活力,Calcein/PI试剂盒法检测细胞活性与细胞毒性,ROS试剂盒法检测细胞氧化应激,RT-qPCR法检测细胞中TNF-α、IL-6、IL-1β的m RNA水平,免疫荧光法检测NLRP3在细胞中的表达水平。结果细胞活力检测结果显示,与NC组相比,HDM组的细胞活力显著降低(P<0.01),HDM+H_(2)组较HDM组细胞活力显著增加(P<0.01)。细胞活性与细胞毒性检测结果显示,与NC组相比,HDM组的红色荧光显著增加,标志细胞死亡率显著升高(P<0.05);HDM+H_(2)组与HDM组相比红色荧光显著减少,标志细胞死亡率显著降低(P<0.05)。ROS检测结果显示,与NC组相比,HDM组绿色荧光显著增加(P<0.01),HDM+H_(2)组与HDM组相比绿色荧光显著减弱(P<0.01)。RT-qPCR检测结果显示,与NC组相比,HDM组中IL-6、IL-1β以及TNF-α的m RNA含量显著增加(P<0.01),与HDM组相比,HDM+H_(2)组中IL-6、IL-1β以及TNF-α的m RNA含量显著降低(P<0.05)。免疫荧光检测结果显示,与NC组相比,HDM组NLRP3的表达明显增加(P<0.001);与HDM组相比,HDM+H_(2)组细胞中NLRP3的表达明显降低(P<0.01)。结论H_(2)可能通过抑制氧化应激,减少促炎因子释放来保护HDM诱导的HNEPC细胞损伤。Objective To investigate the protective effect of hydrogen(H_(2))on oxidative stress induced by house dust mite ex-tract(HDM)in human nasal mucosal epithelial cells(HNEPC)and to speculate on its possible mechanism.Methods HNEPC cells cultured in vitro were randomly divided into 4 groups:NC group(without any treatment),HDM group(10μg/mL HDM to construct AR model in vitro),HDM+H_(2)group,H_(2)group.CCK8 assay was used to detect cell viability,Calcein/PI assay was used to detect cell activity and cytotoxicity,ROS assay was used to detect cellular oxidative stress,RT-qPCR assay was used to detect mRNA levels of TNF-α,IL-6 and IL-1β,and immunofluorescence assay was used to detect the expression level of NLRP3 in cells.Results Compared with the NC group,the cell viability of HDM group was signifi-cantly decreased(P<0.01),and the cell viability of HDM+H_(2)group was significantly increased(P<0.01).The results of cell activi-ty and cytotoxicity detection showed that compared with NC group,the red fluorescence and mortality of marker cells in HDM group were significantly increased(P<0.05).The red fluorescence of HDM+H_(2)group and HDM group was significantly decreased,and the mortality of marker cells was significantly decreased(P<0.05).The ROS detection results showed that compared with the NC group,the green fluorescence of the HDM group was significantly increased(P<0.01),and the green fluorescence of the HDM+H_(2)group was significantly decreased compared with the HDM group(P<0.01).The results of RT-qPCR showed that compared with the NC group,the mRNA contents of IL-6,IL-1βand TNF-αin HDM group were significantly increased(P<0.01),and the mRNA contents of IL-6,IL-1βand TNF-αin HDM+H_(2)group were significantly decreased compared with the HDM group(P<0.05).The results of immu-nofluorescence detection showed that NLRP3 expression was significantly increased in HDM group compared with NC group(P<0.001).Compared with HDM group,the expression of NLRP3 in HDM+H_(2)group was significantly decreased(P<0.01).Conclusion

关 键 词：变应性鼻炎 人鼻黏膜上皮细胞 氢气 氧化应激 保护作用 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]