突发性聋患者糖皮质激素治疗前后外周血单个核细胞Nrf2表达分析

Study of the Expression of Nrf2 in Peripheral Blood Mononuclear Cells From Sudden Deafness Patients Treated with Methylprednisolone

齐慧;戴艳红;后婕;周琼琼;魏先梅;杨烨;佘万东;

1:南京大学医学院附属鼓楼医院耳鼻咽喉科

摘要
目的探讨突发性聋(sudden sensorineural hearing loss,SSNHL)患者体内转录因子NF-E2相关因子2(nuclear factor erythroid2related factor,Nrf2)的变化及其与糖皮质激素(glucocorticoid,GC)抵抗的关系。方法分离30例SSNHL患者治疗前后外周血单个核细胞(peripheral blood mononuclear cells,PBMCs),提取总RNA,采用实时荧光定量PCR(quantitative real time PCR,QRT-PCR)方法检测其PBMCs中Nrf2mRNA的表达量。根据就诊时间及用药情况将这30例SSNHL患者分为初发组(7例)和难治组(23例);难治组又根据听力恢复是否≥15dB分为GC有效组(10例)及抵抗组(13例);选择10例听力正常的志愿者为对照组;比较各组SSNHL患者治疗前后Nrf2mRNA表达量及与GC抵抗的关系。结果 130例患者中,治疗前难治组、初发组Nrf2mRNA表达量分别为0.520±0.049、0.514±0.052,两组间差异无统计学意义(F=0.052,P=0.911),但均低于对照组(0.607±0.106)(F=6.483,P=0.004);2治疗前难治组中GC有效组、抵抗组患耳0.258kHz平均听阈(pure tone average,PTA)分别为81.75±13.28和91.80±13.79dB HL,治疗后分别为54.67±15.65和89.87±13.98dB HL。治疗后GC有效组Nrf2 mRNA表达量(0.560±0.051)较治疗前(0.510±0.028)有所上调(t=3.007,P=0.015),而GC抵抗组Nrf2mRNA表达量(0.519±0.054)较治疗前(0.526±0.060)无明显变化(t=0.485,P=0.636)。结论 Nrf2表达上调或下降可能参与SSNHL的发生发展;难治性SSNHL患者GC抵抗的机制可能与Nrf2低表达相关,上调Nrf2可能改善难治性SSNHL患者对GC的敏感性和预后。
关键词
突发性聋;糖皮质激素;NF-E2相关因子2;外周血单个核细胞
基金项目(Foundation):
国家自然科学基金(81271074);; 江苏省临床医学科技专项(BL2014002);; 江苏省第十批“六大人才高峰项目”(WSN-009);; 南京市国际联合研究项目(2012sd311038)联合资助
作者
齐慧;戴艳红;后婕;周琼琼;魏先梅;杨烨;佘万东;
参考文献

1 Stachler RJ,Chandrasekhar SS,Archer SM,et al.Clinical practice guideline:sudden hearing loss[J].Otolaryngol Head Neck Surg,2012,146:S1.

2 中国突发性聋多中心临床研究协作组.中国突发性聋分型治疗的多中心研究[J].中华耳鼻咽喉头颈外科杂志,2013,48:355.

3 Wei BP,Stathopoulos D,O'Leary S.Steroids for idiopathic sudden sensorineural hearing loss[J].Cochrane Database Syst Rev,2013,7.doi:10.1002/14651858.

4 Schreiber BE,Agrup C,Haskard DO,et al.Sudden sensorineural hearing loss[J].Lancet,2010,375:1203.

5 Nakagawa T,Sakamoto T,Hiraumi H,et al.Topical insulin-like growth factor 1treatment using gelatin hydrogels for glucocorticoidresistant sudden sensorineural hearing loss:a prospective clinical trial[J].BMC Med,2010,8:76.

6 She W,Dai Y,Du X,et al.Hearing evaluation of intratympanic methylprednisolone perfusion for refractory sudden sensorineural hearing loss[J].Otolaryngol Head Neck Surg,2010,142:266.

7 Barnes PJ,Adack IM.Glucocorticoid resistance in inflammatory diseases[J].Lancet,2009,373:1905.

8 Hou J,She W,Du X,et al.Histone deacetylase 2in sudden sensorineural hearing loss patients in response to intratympanic methylprednisolone perfusion[J].Otolaryngol Head Neck Surg,2015,doi:10.177/0194599815606911.

9 后婕,戴艳红,谢利生,等.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[J].听力学及言语疾病杂志,2014,22:559.

10 Malhotra D,Thimmulappa RK,Mercado N,et al.Denitrosylation of HDAC2by targeting Nrf2restores glucocorticosteroid sensitivity in macrophages from COPD patients[J].J Clin Invest,2011,121:4289..

11 Strom J,Xu B,Tian X,et al.Nrf2protects mitochondrial decay by oxidative stress[J].FASEB J,2015,doi:10.1096/fi.14-268904.

12 Biswal S,Thimmulappa RK,Harvey CJ.Experimental therapeutics of Nrf2as a target for prevention of bacterial exacerbations in COPD[J].Proc Am Thorac Soc,2012,9:47.

13 Kim SJ,Park C,Han AL,et al.Ebselen attenuates cisplatin-induced ROS generation through Nrf2activation in auditory cells[J].Hear Res,2009,251:70.

14 中华医学会耳鼻咽喉头颈外科学分会,中华耳鼻咽喉头颈外科杂志编辑委员.突发性聋的诊断和治疗指南[J].中华耳鼻咽喉头颈外科杂志,2015,50:443.

15 Capaccio P,Pignataro L,Gaini LM,et al.Unbalanced oxidative status in idiopathic sudden sensorineural hearing loss[J].Eur Arch Otorhinolaryngol,2012,269:449..

16 Yang CH,Ko MT,Peng JP,et al.Zinc in the treatment of idiopathic sudden sensorineural hearing loss[J].Laryngoscope,2011,121:617.

17 Wiegman CH,Li F,Clarke CJ,et al.A comprehensive analysis of oxidative stress in the ozone-induced lung inflammation mouse model[J].Clin Sci(Lond),2014,126:425.

18 王璐,丁大连,孙虹,等.对内耳氧化抗氧化失衡的反思[J].临床耳鼻咽喉头颈外科杂志,2013,22:965.

19 Chang J,Jung HH,Yang JY,et al.Protective effect of metformin against cisplatin-induced ototoxicity in an auditory cell line[J].J Assoc Res Otolaryngol,2014,15:149.

20 Du Z,Yang Y,Hu Y,et al.A long-term high-fat diet increases oxidative stress,mitochondrial damage and apoptosis in the inner ear of D-galactose-induced aging rats[J].Hear Res,2012,287:15.

21 Hielscher A,Gerecht S.Hypoxia and free radicals:Role in tumor progression and the use of engineering-based platforms to address these relationships[J].Free Radical Biology and Medicine,2015,79:281.

22 Riva C,Donadieu E,Magnan J,et al.Age-related hearing loss in CD/1 mice is associated to ROS formation and HIF target proteins up-regulation in the cochlea[J].Exp Gerontol,2007,2:327.

23 Ito K,Hanazawa T,Tomita K,et al.Oxidative stress reduces histone deacetylase2activity and enhances IL-8gene expression:role of tyrosine nitration[J].Biochem Biophys Res Commun,2004,315:240.

24 Adenuga D,Caito S,Yao H,et al.Nrf2deficiency influences susceptibility to steroid resistance via HDAC2reduction[J].Biochem Biophys Res Commun,2010,403:452.

25 Mercado N,Thimmulappa R,Thomas CM,et al.Decreased histone deacetylase 2impairs Nrf2 activation by oxidative stress[J].Biochem Biophys Res Commun,2011,406:292.