不同剂量白喉毒素对野生型听力成熟小鼠耳蜗结构及听功能的影响

Effects of Different Doses of Diphtheria Toxin on the Cochlear Structure and Auditory Function in Wildtype Auditory-Mature Mice

潘豪来;王际平;冯艳梅;王慧;殷善开;

1:第六人民医院耳鼻喉科研究所上海交通大学附属第六人民医院

摘要
目的观察不同剂量白喉毒素对野生型听力成熟小鼠的耳蜗结构及听功能的影响,为建立白喉毒素受体介导的细胞敲除系统动物模型提供参考。方法 4周龄C57BL/6J小鼠30只随机分为50ng/g组、100ng/g组和对照组,每组10只;其中50ng/g组和100ng/g组小鼠分别单次腹腔注射白喉毒素50ng/g、100ng/g,对照组小鼠单次腹腔注射等量生理盐水,注射后7天观察两组动物的一般情况,并记录小鼠ABR反应阈,观察小鼠内外毛细胞、螺旋神经节神经元。结果在白喉毒素注射后第7天,各组动物情况良好,无明显体重下降,中耳腔无积液。50ng/g组小鼠2、8、32kHz ABR反应阈分别为57.5±2.74、20.83±2.04、45.83±2.04dB SPL,与对照组比较(分别为56.25±2.31、20.0±3.78、49.38±6.78dB SPL)差异无统计学意义;内、外毛细胞损失率分别为0.8%±0.5%、1%±0.6%;对照组分别为0.3%±0.5%、0.4%±0.3%;螺旋神经节神经元SGN密度为39.45±3.65个/105μm2、对照组为41.03±3.73个/105μm2,均未见明显细胞缺失。100ng/g组小鼠2、8、32kHz ABR反应阈分别为85±3.54、63±4.47、90±0dB SPL,较前两组显著升高(t=19.62,P<0.001),内、外毛细胞损失率分别为0.5%±0.1%、10.7%±0.3%,损伤严重,底、中、顶回缺失达10%(t=42.219,P<0.001);SGN密度为25.55±3.66个/105μm2,平均减少38%,与对照组相比差异有显著统计学意义(t=10.985,P<0.001)。结论 100ng/g白喉毒素注射能引起听力成熟野生型C57BL/6J小鼠外毛细胞及螺旋神经元的损伤。
关键词
白喉毒素;耳毒性;白喉毒素受体
基金项目(Foundation):
上海市人才发展基金资助项目(201613)
作者
潘豪来;王际平;冯艳梅;王慧;殷善开;
参考文献

1 Buch T,Heppner FL,Tertilt C,et al.A Cre-inducible diphtheria toxin receptor mediates cell lineage ablation after toxin administration[J].Nature methods,2005,2:419.

2 Saito M,Iwawaki T,Taya C,et al.Diphtheria toxin receptor-mediated conditional and targeted cell ablation in transgenic mice[J].Nat Biotechnol,2001,19:746.

3 Meyer Zu Horste G,Zozulya AL,El-Haddad H,et al.Active immunization induces toxicity of diphtheria toxin in diphtheria resistant mice--implications for neuroinflammatory models[J].J Immunol Methods,2010,354:80.

4 Goldwich A,Steinkasserer A,Gessner A,et al.Impairment of podocyte function by diphtheria toxin--a new reversible proteinuria model in mice[J].Laboratory Investigation,2012,92:1674.

5 Christiaansen AF,Boggiatto P M,Varga SM.Limitations of Foxp3(+)Treg depletion following viral infection in DEREGmice[J].J Immunol Methods,2014,406:58.

6 Narui Y,Minekawa A,Iizuka T,et al.Development of distortion product otoacoustic emissions in C57BL/6J mice[J].International Journal of Audiology,2009,48:576.

7 Konishi H,Ohgami N,Matsushita A,et al.Exposure to diphtheria toxin during the juvenile period impairs both inner and outer hair cells in C57BL/6 mice[J].Neuroscience,2017.DOI:10.1016/j.neuroscience.

8 Song Y,Xia A,Lee HY,et al.Activity-dependent regulation of prestin expression in mouse outer hair cells[J].J Neurophysiol,2015,113:3531.

9 Hu L,Lu J,Chiang H,et al.Diphtheria toxin-induced cell death triggers wnt-dependent hair cell regeneration in neonatal mice[J].J Neurosci,2016,36:9479.

10 Naglich JG,Metherall JE,Russell DW,et al.Expression cloning of a diphtheria toxin receptor:identity with a heparin-binding EGF-like growth factor precursor[J].Cell,1992,69:1051.

11 Mitamura T,Higashiyama S,Taniguchi N,et al.Diphtheria toxin binds to the epidermal growth factor(EGF)-like domain of human heparin-binding EGF-like growth factor/diphtheria toxin receptor and inhibits specifically its mitogenic activity[J].J Biol Chem,1995,270:1015.

12 Cha JH,Brooke JS,Eidels L.Toxin binding site of the diphtheria toxin receptor:loss and gain of diphtheria toxin binding of monkey and mouse heparin-binding,epidermal growth factor-like growth factor precursors by reciprocal site-directed mutagenesis[J].Mol Microbiol,1998,29:1275.

13 Pienkowski M,Ulfendahl M.Differential effects of salicylate,quinine,and furosemide on Guinea pig inner and outer hair cell function revealed by the input-output relation of the auditory brainstem response[J].J Am Acad Audiol,2011,22:104.

14 RR M.Corynebacterium diphtheriae.In:Mandell GL,Bennett JE,Dolin R,editors.Mandell,Douglas,and Bennett's principles and practice of infectious diseases[M].4th edition.New York:Churchill Livingstone,1995.1865~1872.

15 Baxter R,Lewis N,Bohrer P,et al.Sudden-onset sensorineural hearing loss after immunization:a casecentered analysis[J].Otolaryngol Head Neck Surg,2016,155:81.

16 Nuss RC,Evans G.Hearing loss not associated with DTPvaccination or fever[J].Pediatr Neonatol,1993,92:740.

17 Schubert CR,Cruickshanks KJ,Wiley TL,et al.Diphtheria and hearing loss[J].Public Health Rep,2001,116:362.

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白喉毒素耳毒性白喉毒素受体