HDAC2对豚鼠急性听力损失治疗中糖皮质激素抵抗的影响

HDAC2 is Associated with Glucocorticoid Sensitivity in LPS-induced Sudden Hearing Loss in Guinea Pig

周琼琼;佘万东;

1:南京大学医学院附属鼓楼医院耳鼻咽喉科

摘要
目的观察地塞米松(dexamethasone,DEX)加氨茶碱(amionophylline,AMI)治疗脂多糖(lipopolysaccharide,LPS)诱导的豚鼠急性听力损失的疗效及耳蜗组蛋白去乙酰化酶2(histone deacetylase 2,HDAC2)的表达,探讨治疗中HDAC2对糖皮质激素(glucocorticoid,GC)抵抗的影响。方法选取50只健康白色豚鼠,其中10只为对照组,仅双耳鼓阶注入5μl人工外淋巴液(artificial perilymph,AP)(AP组);其余40只鼓阶注入5μl(5mg/ml)的LPS后再随机分为4组,每组10只:模型组(LPS组),无其他处理;LPS+DEX组:术前30min及术后24小时腹腔注射DEX 1mg/kg;LPS+AMI组:术前30min及术后24小时腹腔注射AMI 20mg/kg;LPS+DEX+AMI组:术前30min及术后24小时腹腔注射AMI 20mg/kg及DEX 1mg/kg。所有动物给药前及鼓阶注射术后48小时行ABR检测后取耳蜗,采用免疫荧光法,观察耳蜗基底膜铺片及耳蜗组织冰冻切片中HDAC2的分布及空间定位,用ELISA法测定耳蜗组织中HDAC2的含量。结果 LPS组鼓阶注射LPS后48小时全频听力损失,由低频向高频逐渐加重;LPS+DEX组及LPS+AMI组16、32kHz ABR阈移较LPS组改善明显(<0.05),LPS+DEX+AMI组各频率ABR阈移均低于LPS+DEX组(P<0.05),以16kHz处最显著(P<0.01)。耳蜗基底膜铺片及耳蜗冰冻切片免疫染色结果表明HDAC2广泛分布于耳蜗中,且主要存在于胞核内。8、16、32kHz ABR阈移与耳蜗内HDAC2的含量呈负相关(P<0.05)。结论 AMI可提高豚鼠耳蜗HDAC2的表达,HDAC2可改善治疗中GC抵抗,对提高GC治疗LPS导致的急性听力损失的敏感性有一定的作用。
关键词
组蛋白去乙酰化酶2;糖皮质激素抵抗;听力损失;氨茶碱;豚鼠
基金项目(Foundation):
国家自然科学基金项目(81271074);; 江苏省第十批“六大人才”高峰项目(WSN-009);; 江苏省临床医学科技专项(BL2014002);; 南京市国际联合研究项目(2012sd311038)联合资助
作者
周琼琼;佘万东;
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