摘要
目的 研究肌醇需求酶1(inositol-requiring enzyme 1,IRE1)对顺铂所致小鼠耳蜗毛细胞凋亡的作用。方法 分离生后3~5 d的健康昆明小鼠(500只)耳蜗基底膜并进行体外培养,24 h后随机分为对照组和不同浓度顺铂组(4、8、16和32μg/ml)(每组200条),再培养24 h。应用异硫氰酸荧光素(FITC)染色观察小鼠耳蜗毛细胞的缺失;应用Hoechst 33258和四甲基异硫氰酸罗丹明(TRITC)标记的鬼笔环肽双重荧光染色观察小鼠耳蜗毛细胞的凋亡;并用Western blot检测小鼠耳蜗中p-IRE1α、p-JNK、Bax及caspase-3的蛋白水平。结果 与对照组比较,顺铂组小鼠耳蜗毛细胞缺失、细胞凋亡率以及p-IRE1α、p-JNK、Bax、caspase-3等蛋白水平显著增高(P<0.01),并且p-IRE1α与Bax、caspase-3表达和细胞凋亡率呈正相关(P<0.05)。结论 IRE1可能参与调控顺铂所致小鼠耳蜗毛细胞凋亡。
Objective To investigate the effect of inositol-requiring enzyme 1(IRE1) on cisplatin-induced apoptosis of cochlear hair cells in mouse.Methods The cochlear basilar membranes of 500 healthy Kunming mice(3~5 days old) were isolated and cultured in vitro.After 24 hours,they were randomly divided into control group and cisplatin groups(4 μg/ml,8 μg/ml,16 μg/ml and 32 μg/ml),and cultured for another 24 hours.The loss of cochlear hair cells was observed by fluorescein isothiocyanate(FITC) staining.The apoptosis of cochlear hair cells was observed by TRITC and Hoechst 33258 fluorescence staining.Western blot was used to detect the protein levels of p-IRE1α,p-JNK,Bax and caspase-3 in each group.Results Compared with control group,the hair cell loss and apoptosis rate of hair cells and the protein levels of p-IRE1α,p-JNK,Bax and caspase-3 were significantly increased in cisplatin groups(P<0.01).Furthermore,the expression of p-IRE1α was positively correlated with the expression of Bax and caspase-3 and with the apoptosis rate of hair cells(P<0.05).Conclusion IRE1 may be involved in the regulation of cisplatin-induced apoptosis in cochlear hair cells.
作者
张晓
王爱梅
Zhang Xiao;Wang Aimei(Department of Anatomy,Henan Vocational College of Nursing,Anyang,455000,China;不详)
出处
《听力学及言语疾病杂志》
CAS
CSCD
北大核心
2024年第2期150-154,共5页
Journal of Audiology and Speech Pathology
基金
辽宁省“兴辽英才计划”项目资助(XLYC2002023)。
关键词
肌醇需求酶1
顺铂
耳蜗
凋亡
Inositol-requiring enzyme 1
Cisplatin
Cochlea
Apoptosis
