难治性突发性聋患者外周血单个核细胞内HDAC2表达与其预后的相关性分析

发布时间：2019-08-01 07:43

【摘要】：目的探讨组蛋白去乙酰化酶2(histone deacetylase 2,HDAC2)水平与突发性聋(sudden sensorineural hearing loss,SSNHL)患者糖皮质激素(glucocorticoid,GC)抵抗的关系。方法选择难治性SSNHL患者20例,所有患耳鼓室内灌注甲强龙20mg/d,连续10天,同时辅以全身静脉滴注营养神经及改善微循环药物。对照组为10例正常听力志愿者。分离所有研究对象的外周血单个核细胞(peripheral blood mononuclear cell,PBMC),提取细胞核蛋白以及总RNA,分别用HDAC2活性检测试剂盒以及real time PCR的方法,检测受试者PBMC中HDAC2的蛋白活性以及HDAC2mRNA的表达情况,并随访SSNHL患者听力,根据听力恢复情况将SSNHL患者分为GC有效组和GC无效组,探讨HDAC2水平与GC抵抗的关系。结果 20例SSNHL患者中,GC有效8例,GC无效12例,治疗前GC有效组及无效组患耳PTA分别为75.21±9.44和80.98±12.73dB HL,治疗后分别为40.83±14.37和77.92±12.25dB HL;治疗前突聋患者HDAC2蛋白活性OD值(GC有效组0.664±0.022,GC无效组0.659±0.043)显著低于对照组(0.726±0.024),而HDAC2mRNA表达量(GC有效组0.079±0.023,GC无效组0.083±0.047)显著高于对照组(0.035±0.022),治疗后,两组患者HDAC2mRNA表达量(GC有效组0.132±0.041,GC无效组0.132±0.048)较治疗前升高,GC有效组患者HDAC2蛋白活性(0.730±0.033)上升并接近正常水平,而GC无效组患者HDAC2蛋白活性(0.647±0.013)无明显变化。结论难治性SSNHL患者对GC不敏感可能与HDAC2蛋白活性降低有关,GC可以促使HDAC2mRNA的表达上调,GC治疗无效的患者体内可能存在某种机制,对HDAC2进行翻译后修饰,使HDAC2蛋白活性降低,从而抑制组蛋白去乙酰化,导致其对激素治疗不敏感。
[Abstract]:Objective to investigate the relationship between histone deacetylase 2 (histone deacetylase 2, HDAC 2) and glucocorticoid (glucocorticoid,GC) resistance in patients with sudden deafness (sudden sensorineural hearing loss,SSNHL). Methods 20 patients with refractory SSNHL were enrolled in this study. All patients were infused with prednisolone 20 mg / d for 10 days, supplemented by systemic intravenous infusion of nutritive nerve and drugs to improve microcirculation. There were 10 normal hearing volunteers in the control group. (peripheral blood mononuclear cell,PBMC was isolated from peripheral blood mononuclear cells (PBMCs). Nuclear protein and total RNA, were extracted from peripheral blood mononuclear cells (PBMCs). The protein activity of HDAC2 and the expression of HDAC2mRNA in PBMC were detected by HDAC2 activity kit and real time PCR, respectively. the hearing of SSNHL patients was followed up. SSNHL patients were divided into GC effective group and GC ineffective group according to hearing recovery to explore the relationship between HDAC2 level and GC resistance. Results among 20 patients with SSNHL, GC was effective in 8 cases and GC was ineffective in 12 cases. The PTA of ears in GC effective group and ineffective group were 75.21 卤9.44 and 80.98 卤12.73dB HL, respectively after treatment, 40.83 卤14.37 and 77.92 卤12.25dB HL;, respectively. The activity of HDAC2 protein in patients with sudden deafness before treatment (GC effective group 0.664 卤0.022, GC ineffective group 0.659 卤0.043) was significantly lower than that in control group (0.726 卤0.024), while HDAC2mRNA expression (GC effective group 0.079 卤0.023, GC ineffective group 0.083 卤0.047) was significantly higher than that in control group (0.035 卤0.022). After treatment, the expression of HDAC2mRNA in the two groups (GC effective group 0.132 卤0.041, GC ineffective group 0.132 卤0.048) was significantly higher than that before treatment. The activity of HDAC2 protein in GC effective group (0.730 卤0.033) increased and approached the normal level, but there was no significant change in HDAC2 protein activity in GC ineffective group (0.647 卤0.013). Conclusion the insensitivity to GC in patients with refractory SSNHL may be related to the decrease of HDAC2 protein activity. GC can up-regulate the expression of HDAC2mRNA. There may be some mechanism in patients with ineffective GC treatment. Post-translation modification of HDAC2 can reduce the activity of HDAC2 protein, thus inhibiting histone deacetylation and insensitive to hormone therapy.
【作者单位】：南京大学医学院附属鼓楼医院耳鼻咽喉科;
【基金】：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
【分类号】：R764.437

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