激素抵抗对突发性聋治疗效果影响的临床观察

摘要

Objective To investigate the correlation between the proliferation inhibition effect of glucocorticoid (GC) on peripheral blood mononuclear cell (PBMC) and the pure tone average (PTA) improvement in SSNHL patients.Methods Sixty inpatients with SSNHL were included from July 2013 to October 2015 in Nanjing Drum Tower Hospital, Medical School of Nanjing University.Peripheral venous blood was collected before receiving treatment, then the PBMC was isolated for GC proliferation inhibition.PBMCs of each patient were cultivated into 4 groups: Group A: PBMCs+Medium;Group B: PBMCs+Medium+lipopolysaccharide (LPS, 1 μmol/L);Group C: PBMCs+Medium+LPS+Dexamethasone;Group D: Medium.PBMCs were maintained in a humidified 5% CO2 atmosphere at 37°C and were observed after 24 hours.5-diphenyltetrazolium bromide (MTT) was used to measure PBMC proliferation inhibition rate.The PBMC proliferation inhibition rates were calculated according to the absorbance at 490 nm wavelength under a microtiter plate reader.Independent sample t tests of PBMC proliferation inhibition rate were performed between different groups.x2 tests were performed between gender, affected ear side, accompanied by vertigo or not, audiometric curve, time period from onset to treatment, PBMC proliferation inhibition rate and the improvement of pure tone average (PTA).Linear correlation analyses were performed between PBMC proliferation inhibition rate, the time period from onset to treatment and the hearing improvement.Results The proliferation inhibition effect of GC on PBMC varied significantly among patients.The PBMC proliferation inhibition rate in GC insensitive group was lower than that in GC sensitive group (26.72%±21.82% vs 64.44%±25.48%, t=6.113, P<0.05).The PBMC proliferation inhibition rate in refractory group was lower than that in initial group (40.93%±28.57% vs 57.04%±31.19%, t=2.035, P=0.046).There was no statistical significance between gender, affected ear side, accompanied by vertigo or not, audiometric curve and the hearing improvement (x2 value was 2.320, 0.031, 2.143, 0.106, respectively, all P>0.05).Both in initial group and refractory group, the linear correlation analyses showed a significant positive correlation between PBMC proliferation inhibition rate and the PTA improvement (r value was 0.615, 0.657, respectively, all P<0.05), as well as a significant negative correlation between time period from onset to treatment and the PTA improvement(r value was-0.542, 0.370, respectively, all P<0.05).Conclusions The proliferation inhibition rate of PBMC in vitro by GC is correlated with patients′ hearing improvement.The proliferation inhibition test might be used to predict the sensitivity to GC treatment and be helpful for individualized treatment of SSNHLin clinical practice.%目的 观察突发性聋(sudden sensorineural hearing loss,SSNHL)患者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)增殖抑制情况与其听力预后的关系.方法 纳入2013年7月至2015年10月于南京大学医学院附属鼓楼医院耳鼻咽喉头颈外科住院接受治疗的SSNHL患者60例,在治疗前空腹抽取外周血,提取PBMC进行体外增殖糖皮质激素(glucocorticoid,GC)抑制试验.细胞培养分为4组:A组仅含有细胞及培养基而不添加药物,为对照组;B组加脂多糖(1 μmol/L,下同)刺激PBMC增殖;C组同时加入脂多糖和地塞米松(10-6 mol/L);D组为不添加细胞仅有培养基的空白组.在37℃、5%CO2恒温箱培养24 h后,观察细胞生长情况,并采用噻唑蓝染色法检测其在490 nm波长处的吸光度来反映细胞的增殖抑制情况.不同组间增殖抑制率的比较采用独立样本t检验.患者性别、患耳侧别、是否伴有眩晕、听力分型、发病至就诊时间、PBMC增殖抑制率等各因素分别与受损频率平均听阈(pure tone average,PTA)改善值的关系采用卡方检验.患者发病至就诊的时间和增殖抑制率与PTA改善值之间的相关性采用Linear线性回归分析.将P<0.05设为差异有统计学意义.结果 培养24 h后,所有患者的PBMC均可被地塞米松抑制,但抑制程度有很大差异.GC无效组的PBMC增殖抑制率显著低于GC有效组(26.72%±21.82%比64.44%±25.48%,t=6.113,P<0.05);难治组的PBMC增殖抑制率显著低于初诊组(40.93%±28.57%比57.04%±31.19%,t=2.035,P=0.046).患者的性别、患耳侧别、发病时是否伴有眩晕及听力曲线对患者听力预后影响的差异不具有统计学意义(x2值分别为2.320、0.031、2.143、0.106,P值均>0.05).在初诊组和难治组患者中,PTA改善值与PBMC增殖抑制率之间均存在正相关关系(r值分别为0.615、0.657,P值均<0.05),PTA改善值与发病至就诊时间之间均存在负相关关系(r值分别为-0.542、-0.370,P值均<0.05).结论 SSNHL患者的PBMC体外增殖抑制情况与听力预后相关,可用于了解患者对GC的敏感性,为预测预后提供参考.