大鼠耳蜗一氧化氮合酶的分布和表达

Distribution and Expression of NOS in Rat Cochlea

潘世恺;角南贵司子;山根英雄;王正敏

1:复旦大学附属眼耳鼻喉科医院耳鼻咽喉科

2:曰本国大阪市立大学医学部耳鼻咽喉科学教研室

3:曰本国大阪市立大学医学部耳鼻咽喉科学教研室

4:复旦大学附属眼耳鼻喉科医院耳鼻咽喉科 上海200031

摘要
目的 本实验先用组织化学法 ,通过观察还原型辅酶Ⅱ -黄递酶 (NADPH -黄递酶 )的活性了解一氧化氮合酶 (nitricoxidesynthase ,NOS)在大鼠耳蜗内分布。再用亲合免疫细胞组织化学技术 ,研究大鼠耳蜗内神经元型NOS(neuronalNOS ,nNOS)与内皮型NOS(endothelialNOS ,eNOS)的表达。方法 组化组大鼠耳蜗切片用辅酶Ⅱ孵育液在 37℃条件下孵育 1小时。免疫组化组大鼠耳蜗切片经消除内源性过氧化物酶 ,3%山羊正常血清封闭非正常结合点后 ,用兔抗nNOS抗体、兔抗eNOS抗体 ,室温下孵育 6 0分钟 ,再用生物素标记的山羊抗兔第二抗体孵育、滴加ABC试剂 ,以DAB试剂显色。结果 大鼠耳蜗血管球内皮细胞有明显NADPH -黄递酶活性 ,血管纹及螺旋神经节细胞也有NADPH -黄递酶活性反应。大鼠耳蜗内、外毛细胞、螺旋神经节细胞nNOS、eNOS的表达呈阳性。血管纹细胞处有阳性nNOS、eNOS的表达。耳蜗血管球的内皮细胞无nNOS的表达 ,但eNOS的表达呈强阳性。结论 由nNOS及eNOS合成的NO在维持耳蜗正常神经传导及耳蜗毛细血管张力和正常血液供应中起着重要作用
关键词
一氧化氮合酶;一氧化氮;还原型辅酶Ⅱ-黄递酶;免疫组化;耳蜗;大鼠
基金项目(Foundation):
作者
潘世恺;角南贵司子;山根英雄;王正敏
参考文献

1 BrechtelsbauerPB ,NuttalAl,MillerJM .Basalnitricoxideproduc tioninregulationofcochlearbloodflow[J].HearRes,1994,77:38. 

2 FessendenJD ,SchachtJ.Localizationofsolubleguanylatechclaseac tivityintheguineapigcochleasuggestsinvolvementinregulationofbloodflowandsupportingcellphysilogy[J].JHistochemCytochem,1997,45:1401.

3 GosepathK ,GathI,MaurerJ ,etal.Characterizationofnitricoxidesynthaseisoformsexpressedindifferentstructuresoftheguineapigcochlea[J].BrainRes,1997,747:26. 

4 FessendenJD ,Coling,DE ,SchachtJ.Detectionandcharacterizationofnitricoxidesynthaseinthemammaliancochlea[J].BrainRes,1994,668:9. 

5 潘世恺,角南贵司子,山根英雄,等.豚鼠耳蜗一氧化氮合酶的分布[J].听力学与言语疾病杂志,2001,9:19. 

6 潘世恺,角南贵司子,山根英雄,等.一氧化氮合酶在小鼠耳蜗的表达[J].复旦学报(医学科学版),2001,28:220. 

7 MatsumotoT ,NakaneM ,PollockJS ,etal.Acorrelationbetweensol ublebrainnitricoxidesynthaseandNADPH -diaphoraseactivityisonlyseenafterexposureofthetissuetofixative[J].NeurosciLett,1993,155:61. 

8 LowensteinCJ,DinermanJL ,SnyderSH .Nitricoxide:aphysiologicmessenger[J].AnnInternMed,1994,120:227. 

9 KonishiK ,YamanaH ,IguchiH ,etal.Localsubstancesregulatingcochlearbloodflow[J].ActaOtolaryngol(Stockh),Suppl,1998,538:40. 

10 RenT ,BrechtelsbauerPB ,MillerJM ,etal.CochlearbloodflowmeasuredbyaveragedlaserDopplerflowmetry(ALDF)[J].HearRes,1994,77:200. 

11 HauptH ,ScheibeF ,RebentischE .Improvementininnerearbloodflowbynitricoxidefollowingexperimentallyinducedcochlearthrom bosisinanesthetizedguineapigs[J].EurArchOtorhinolaryngol,1998,255:334. 

12 TodaK ,KitamuraY ,OkamuraT .Neuralmechanismofhypertensionbynitricoxidesynthaseinhibitorindogs[J].Hypertension,1993,21:3.

13 DuranteW ,KrollMH ,VanhouttePM ,etal.Endothelium-derivedrelaxingfactorinhibitsthrombin-inducedplateletaggregationbyin hibitingplateletphospholipaseC[J].Blood,1992,79:110. 

14 AmaeeFR ,ComisSD ,OsborneMP .Possibleinvolvementofnitricoxideinthesensorineuralhearinglossofbacterialmeningitis[J].Ac taOtolaryngol(Stockh),1997,117:329. 

15 TabuchiK ,KusakariJ,ItoZ .Effectofnitricoxidesynthaseinhibitoroncochleardysfunctioninducedbytransientlocalanoxia[J].ActaOtolaryngol(Stockh),1999,119:179. 

16 PaiN ,ZsanskiCJ,GregoryCW .Sodiumnitroprusside/Nitricoxidecausesapoptosisinspiralganglioncells[J].Otolaryngology-HeadandNeckSurgery,1998,119:323. 

17 RuanRS ,LeongSK ,YeohKH .Ototoxicityofsodiumnitroprusside[J].HearingResearch,1997,114:169.

18 RuanRS ,LeongSK ,YeohKH .Ototoxicityofsodiumnitroprussideisnotduetonitricoxide[J].Exp-Neurol,1999,158:192.