摘要
目的:探究核受体亚家族4A组成员1(nuclear receptor subfamily 4 group A member 1,Nr4a1)Nr4a1激动剂胞孢子酮B(cytosporone B,Csn-B)对小鼠噪声暴露后听力损失的治疗作用。方法:采用双氧水刺激HEI-OC1毛细胞系的方法构建氧化应激细胞模型;通过实时荧光定量PCR(quantitative real-time PCR,q PCR)检测细胞中Nr4a1的mRNA表达水平;分别通过细胞计数试剂盒(cell counting kit-8,CCK8)及流式细胞术的方法检测细胞活力和细胞凋亡水平以评估Csn-B预处理后经双氧水刺激的细胞状态。构建小鼠噪声性听力损失模型,运用qPCR和免疫荧光技术检测噪声暴露后Nr4a1在小鼠耳蜗中的表达;通过检测听性脑干反应(auditory brainstem response,ABR)评估噪声暴露后以及Csn-B连续治疗13 d后小鼠听力情况。结果:双氧水刺激后HEI-OC1毛细胞中Nr4a1表达上升,细胞活力显著下降,凋亡水平显著升高;Csn-B预处理HEI-OC1毛细胞经双氧水刺激,细胞活力显著高于对照组而凋亡水平则显著低于对照组。在体研究结果显示,噪声暴露后小鼠听力显著降低,Nr4a1在小鼠耳蜗中的表达水平显著升高。噪声暴露后经Csn-B治疗小鼠听力得到改善,主要表现为Click-ABR以及Tone Burst-ABR(4000、8000Hz处)阈值下降。结论:Nr4a1激动剂Csn-B增强内耳毛细胞对氧化应激损伤的抵御能力,部分改善噪声暴露后的小鼠听力。
Objective:To investigate the therapeutic effects of the nuclear receptor subfamily 4 group A member 1(Nr4a1)agonist cytosporone B(Csn-B)on noise-induced hearing loss in mice.Methods:The HEI-OC1 outer hair cell line was subjected to hydrogen peroxide stimulation to establish an oxidative stress cell model.PCR was utilized to assess Nr4a1 expression in the cells.Cell viability was measured using CCK8,and flow cytometry was employed to evaluate apoptosis in cells pre-treated with Csn-B before hydrogen peroxide stimulation.By exposing mice to noisy audio,a model for noise-induced hearing loss was established,and PCR and immunofluorescence techniques were employed to detect Nr4a1 expression in the cochlea following noise exposure.ABR testing was conducted to assess mouse hearing both after noise exposure and following 13 days of continuous Csn-B treatment.Results:Nr4a1 expression increased in HEI-OC1 cells after hydrogen peroxide stimulation,accompanied by a significant decrease in cell viability and an increase in apoptosis.Pre-treatment with Csn-B enhanced cell viability and reduced apoptosis in HEI-OC1 cells exposed to hydrogen peroxide compared to the control group.In vivo studies revealed an increase in Nr4a1 expression in the cochlea of mice after noise exposure,associated with a decline in auditory function.Significant hearing recovery was observed after Csn-B treatment,evidenced by decreased thresholds in Click-ABR and Tone Burst-ABR(at 4000 and 8000Hz).Conclusion:The Nr4a1 agonist Csn-B enhances cellular resilience to oxidative stress and partially rescues noise-induced hearing loss in mice.
作者
苏波
赖彦冰
王晓迪
褚汉启
冰丹
SU Bo;LAI Yanbing;WANG Xiaodi;CHU Hanqi;BING Dan(Department of Otolaryngology-Head and Neck Surgery,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)
出处
《神经损伤与功能重建》
2024年第5期249-255,共7页
Neural Injury and Functional Reconstruction
基金
湖北省重点研发计划(突发性耳聋预后预测与精准分型诊疗智能协同体系构建,No.2022BC A006)
武汉市知识创新专项(婴幼儿耳聋筛查防治体系构建及遗传机制研究,No.2022022101015011)
同济医院科研基金重点项目(慢性睡眠剥夺活化内耳巨噬细胞经由NLRP3/IL-1β/IL1R导致听力损失,No.2023A01)。
关键词
Nr4a1
Csn-B
噪声性耳聋
氧化应激
nuclear receptor subfamily 4 group A member 1
cytosporone B
noise-induced hearing loss
oxidative stress
