豚鼠内淋巴囊上皮细胞Na^＋,K^＋—ATP酶不同？…

目的 进一步研究同肉淋巴囊上皮细胞Ｎａ＾＋，Ｋ＾＋－ＡＴＰ酶不同β亚基的表达。方法 采用豚鼠内淋巴囊冰冻切片、原位杂交的方法检测Ｎａ＾＋，Ｋ＾＋－ＡＴＰ酶不同β亚基ｍＲＮＡ在淋巴囊上皮细胞中的表达。结果 在内淋巴囊上皮细胞胞浆内可见Ｎａ＾＋，Ｋ＾－ＡＴＰ酶不同β亚基的阳性颗粒，β１亚基表达较弱，β２亚基表达较强。结论 结合其他报道，内淋巴囊上皮细胞具有多种亚基异构体组成的Ｎａ＾＋，Ｋ＾＋－ＡＴＰ酶     

Na^＋—D^＋—ATP酶活性在豚鼠内淋巴囊的细胞化学定位

Ｎａ＾＋－Ｋ＾＋－ＡＴＰ酶在Ｎａ＾＋－Ｋ＾＋离子主动性跨上皮转运过程中起重要作用。应用对硝基苯酚磷酸（ｐ－ＮＰＰ）为底物的柠檬酸铅一步法Ｋ＾＋－ＮＰＰ酶电细胞化学技术，观察了反映Ｎａ＾＋－Ｋ＾＋－ＡＴＰ酶活性的Ｋ＾＋－ＮＰＰ酶在豚鼠内淋巴囊的超微结构定位分布。结果：在透射电镜下观察，内淋巴囊超薄切片上的Ｄ＾＋－ＮＰＰ酶活性反应产物仅分布于上皮细胞底侧面胞膜的胞浆一侧，向上一直延续至上皮细胞间的紧密     

一氧化氮在豚鼠耳蜗作用的实验研究

为探讨ＮＯ在内耳的作用，采用外淋巴给药途径，观察一氧化氮气体（ＮＯ）、Ｌ－精氨酸、硝普钠及一氧化氮合酶（ＮＯＳ）拮抗剂Ｎ－甲基－Ｌ－精氨酸对耳蜗蜗内电位（ＥＰ）、复合动作电位（ＣＡＰ）及耳蜗微音器电位（ＣＭ）的影响。结果表明，Ｎ－甲基－Ｌ－精氨酸可以使ＥＰ减小５０％，ＣＡＰ振幅降低３３％及ＣＭ振幅略有降低，在此基础上，用Ｌ－精氨酸外淋巴灌流可以逆转Ｎ－甲基－Ｌ－精氨酸所致的改变。ＮＯ持外淋巴缓释能     

豚鼠膜迷路积水模型耳蜗Ca^2＋ATP酶的变化

目的 了解Ｃａ＾２＋－ＡＴＰ酶在耳蜗活动位点及膜迷路积水后耳蜗Ｃａ＾２＋－ＡＴＰ酶的变化。方法 选成年健康、Ｐｒｅｙｅｒ反射正常豚鼠１４只，左耳装圆窗电极一阻塞肉淋巴，经声反应阈（ＣＡＰ阀值）证明膜迷路积水形成；右侧为对照耳。用枸橼酸铅细胞组化法测定Ｃａ＾２＋－ＡＴＰ酶，在透射电镜下Ｃａ＾２＋－ＡＴＰ酶以磷酸铅黑色颗粒显示。结果 对照耳Ｃａ＾２＋－ＡＴＰ酶活动部位在蜗管前庭膜内淋巴侧、内外毛细胞皮     

一氧化氮在豚鼠耳蜗作用的实验研究

为探讨ＮＯ在内耳的作用，采用外淋巴给药途径，观察一氧化氮气体（ＮＯ）、Ｌ－精氨酸、硝普钠及一氧化氮合酶（ＮＯＳ）拮抗剂Ｎ－甲基－Ｌ－精氨酸对耳蜗蜗内电位（ＥＰ）、复合动作电位（ＣＡＰ）及耳蜗微音器电位（ＣＭ）的影响。结果表明，Ｎ－甲基－Ｌ－精氨酸可以使ＥＰ减小５０％，ＣＡＰ振幅降低３３％及ＣＭ振幅略有降低，在此基础上，用Ｌ－精氨酸外淋巴灌流可以逆转Ｎ－甲基－Ｌ－精氨酸所致的改变。ＮＯ持续外淋巴缓释能使Ｎ－甲基－Ｌ－精氨酸导致的ＥＰ、ＣＡＰ及ＣＭ的改变恢复，并超过正常，随之出现快速下降。外淋巴灌流硝普钠后，ＥＰ、ＣＡＰ及ＣＭ短暂升高后逐渐下降，并维持在较低水平。ＣＡＰＮ１波及ＣＭ潜伏期的变化规律与其振幅的变化规律基本一致。结果提示，ＮＯ在生理条件下维持内耳功能，可能参与耳蜗毛细胞微机械特性及敏感性的调节，过量表达可以产生耳蜗毒性     

豚鼠壶腹嵴毛细胞内钙离子活动及ATP的影响

目的 探讨离体前庭毛细胞（ＶＨＣ）去极化时细胞内钙离子活动特征及内耳活性物质ＡＴＰ对离体ＶＨＣ内钙离子浓度的影响及其机制。方法 采用酶孵育后机械分离法分离豚鼠壶腹嵴ＶＨＣ,６μｍｏｌ／Ｌ钙荧光探针Ｆｌｕｏ－３染色,利用激光扫描共聚焦显微镜（ＬＳＣＭ）记录静息状态下、加入高Ｋ＾＋液及不同浓度ＡＴＰ时代表ＶＨＣ内［Ｃａ＾２＋ｉ的荧光强度相对值的变化。结果 细胞外有Ｃａ＾２＋存在时,７５ｎｍｏｌ／ＬＫ＾     

庆大霉素对豚鼠内淋巴囊和血管纹Na^＋—K^＋—ATP酶活性的影响

应用酶细胞化学技术和计算机图像分析，对豚鼠连续肌注庆大霉素２－１５ｄ后内淋巴囊上皮细胞和血管纹边缘细胞的Ｎａ＾＋－Ｋ＾＋－ＡＴＰ酶活性变化进行了定量观察。结果发现，与正常对照组相比，用药２ｄ后两类细胞的Ｎａ＾＋－Ｋ＾＋－ＡＴＰ酶活性反应产物没有明显变化，但连续用药５ｄ后二者的产物均明显减少，１０ｄ和１５ｄ仍呈继续轻微减少趋势。表明庆大霉素既可抑制血管纹边缘细胞Ｎａ＾＋－Ｋ＾＋－ＡＴＰ酶活性，降低其     

Na~＋－K~＋－ATP酶活性在豚鼠内淋巴囊的细胞化学定位

Ｎａ￣＋－Ｋ￣＋－ＡＴＰ酶在Ｎａ￣＋－Ｋ￣＋离子主动性跨上皮转运过程中起重要作用。应用对硝基苯酚磷酸（ｐ－ＮＰＰ）为底物的柠檬酸铅一步法Ｋ￣＋－ＮＰＰ酶电镜细胞化学技术，观察了反映Ｎａ￣＋－Ｋ￣＋－ＡＴＰ酶活性的Ｋ￣＋－ＮＰＰ酶在豚鼠内淋巴囊的超微结构定位分布。结果：在透射电镜下观察，内淋巴囊超薄切片上的Ｋ￣＋－ＮＰＰ酶活性反应产物仅分布于上皮细胞底侧面胞膜的胞浆一侧，向上一直延续至上皮细胞间的紧密连接水平，内淋巴腔面胞膜未见反应产物。提示丰富的Ｎａ￣＋－Ｋ￣＋－ＡＴＰ酶活性特征性地分布在内淋巴囊上皮细胞底侧面胞膜，表明Ｎａ￣＋－Ｋ￣＋－ＡＴＰ酶在内淋巴囊上皮Ｎａ￣＋－Ｋ￣＋离子交换过程中的重要地位。     

窒息对不同龄豚鼠耳蜗生物电的影响

应用微电极技术，观测内耳实验中常用两组不同龄豚鼠及恢复供氧过程中蜗内电位（ＥＰ）、微音电位（ＣＭ）、总和电位（ＳＰ）的变化，发现大龄豚鼠于窒息时ＥＰ下降较小龄豚鼠慢，＋ＳＰ的峰值大，恢复供氧后ＥＰ、ＣＭ的回升快，－ＳＰ的峰值亦大，提示大龄鼠的血管纹、外毛细胞、内毛细胞均较小龄鼠成熟。     

不同内耳组织抗原免疫致自身免疫性感音神经性聋的研究

目的：探讨不同内耳组织抗原免疫所致内耳主要病理损伤部位和听力障碍类型。方法：采用同种螺旋韧带（ＳＬ）、基底膜（ＢＭ）、螺旋神经节（ＳＧ）组织抗原免疫豚鼠，观察内耳组织病理改变和听觉功能变化。结果：ＳＬＡｇ和ＢＭＡｇ免疫组主要表现耳蜗微音器电位阈值升高和复聪现象，以及蜗管内和血管纹的免疫炎性病理改变；ＳＧＡｇ免疫组主要表现听神经复合动作电位阈值升高和幅值降低，内耳病理变化主要位于蜗轴血管及周围和ＳＧ     

同种内耳抗原免疫致聋豚鼠的子代内耳功能和形态学观察

目的观察自身免疫性感音神经性聋(autoimmune sensorineural hearing loss，ASHL)母鼠所产子代内耳听觉和前庭生理功能、病理形态学的变化，初步探讨母鼠体内针对内耳组织抗原免疫反应是否可以造成子代的内耳损伤。方法同种内耳抗原(homogeneous inner ear antigens，HIEAg)持续免疫孕豚鼠，采用耳蜗电图(包括听神经复合动作电位)、总和直流电位、耳蜗微音器电位和眼震电图仪(记录自发性和冷热空气试验)测试母鼠和子鼠的听觉和前庭功能，并检测针对HIEAg的血清特异性体液和细胞免疫反应，采用火棉胶切片和HE染色，光镜观察内耳病理组织学改变。结果ASHL母鼠所产子鼠中，部分(3／7)出现听觉损伤，并发现其血清中特异性抗体水平升高，内耳出现免疫炎性病理损伤。非ASHL母鼠和对照组母鼠所产子代未见明显异常。结论ASHL雌鼠所产子代可出现感音神经性聋，其内耳损伤和功能障碍极可能与针对内耳组织的自身免疫反应(尤其是体液免疫)有关。     

Autoimmune inner ear disease: a review of basic mechanisms and clinical correlates

Otolaryngologists have long sought to identify causes of sensorineural hearing loss that might be reversed by medical treatment. One such entity has become known as autoimmune inner ear disease. The potential improvement in auditory function in these patients subsequent to immunosuppressive therapy has created a desire in clinicians to better understand this disease. This paper begins by reviewing the basic concepts of autoimmunity. The experimental and clinical data concerning autoimmune inner ear disease are then described and analyzed. Finally, conclusions are drawn concerning our current state of understanding of this disease process.     

Autoimmune mouse antibodies recognize multiple antigens proposed in human immune-mediated hearing loss.

HYPOTHESIS: Autoimmune diseased mice with hearing loss will have autoantibodies against the various cochlear antigens proposed in clinical autoimmune inner ear disease. BACKGROUND: Serum antibodies of patients with hearing loss recognize several proteins that are proposed as possible antigenic targets in the ear. This often leads to a clinical diagnosis of autoimmune inner ear disease, although it is not clear how these antibodies cause inner ear disease. Therefore, to better understand the relationship of autoantibodies and ear disease, an examination was made of serum autoantibodies in the MRL/MpJ-Fas(lpr) autoimmune mouse with hearing loss. Similar antibody patterns in the mouse would provide an animal model in which to investigate potential autoimmune mechanisms of this clinical ear disorder. METHODS: Sera from MRL/MpJ-Fas(lpr) autoimmune mice and normal C3H mice were tested by the enzyme-linked immunosorbent assay technique for reactivity against various reported cochlear antigens: heat shock protein 70 (bovine, human, bacterial), laminin, heparan sulfate proteoglycan, cardiolipin, and collagen types II and IV. RESULTS: The autoimmune mouse sera showed significantly greater antibody reactivity against all of the antigens when compared with normal mouse sera. CONCLUSIONS: Serum antibodies from autoimmune mice recognized several putative autoantigens reported for patients with hearing loss, suggesting that comparable antigen-antibody mechanisms might be operating. However, the recognition of multiple antigens did not identify any one as being the specific target in autoimmune hearing loss. The correlation of antibodies in the MRL/MpJ-Fas(lpr) autoimmune mouse and human studies indicates this animal model should aid further investigations into potential cochlear antigens in autoimmune hearing loss.     

热休克蛋白70抗体检测对自身免疫性聋诊断的意义

目的 分析自身免疫性聋的听力学特点，探讨用免疫印迹法(Western blot)检测热休克蛋白70抗体在自身免疫性耳聋诊断中的意义。方法自身免疫性聋14例28耳，其它感音神经性聋8例16耳，正常对照6例12耳，采用免疫印迹法检测各组受试者血清中的热休克蛋白70抗体，并分别作临床听力学检查。结果自身免疫性耳聋者纯音听力图多数为平坦型，TEOAE和／或DPOAE未引出，听性脑干反应多数正常，轻、中度耳聋者可引出镫骨肌声发射，该组8例(57．1％)热休克蛋白70抗体阳性。其它感音神经性聋者纯音听力图多数为高频减退型和平坦型，TEOAE和DPOAE未引出，重度耳聋者ABR未引出，轻、中度耳聋者可引出镫骨肌声发射，1例(12．5%)热休克蛋白70抗体阳性。正常对照组1例(16．7％)热休克蛋白70抗体阳性。结论免疫印迹法检测热休克蛋白70抗体对自身免疫性耳聋诊断有参考价值，对该病的确诊还应结合临床听力学特点。     

Female MRL.MpJ-Fas(lpr) autoimmune mice have greater hearing loss than males

Although women make up approximately 60-70% of the patients with autoimmune hearing loss, little is known about the impact of gender on this cochlear disease. To explore this relationship of gender and autoimmune inner ear disease, an evaluation was made of cochlear function in male and female autoimmune MRL.MpJ-Fas(lpr) mice. Autoimmune disease and hearing loss onset occur at 3-4 months of age, so mice were tested with auditory brainstem response audiometry at 3, 6, and 9 months of age to identify potential gender differences in thresholds. Sera also were analyzed for differences in the autoimmune factors of immune complexes, anti-nuclear antibodies, and hematocrits. By 9 months of age the surviving mice showed a dramatic gender difference. Female mice had thresholds 25-45 dB higher than males at 4, 8, and 16 kHz, although male thresholds at 32 kHz had risen sufficiently to be statistically similar to those for females. No gender differences were seen in any of the systemic autoimmune factors. These findings of worse hearing in female autoimmune mice parallel a reported female preponderance in clinical immune hearing disorders. This potential gender influence in autoimmune inner ear disease must be better understood for effective evaluation and treatment of this disorder.     

Hearing thresholds in patients affected by rheumatoid arthritis

The aim of the study was to evaluate hearing thresholds in 38 patients with rheumatoid arthritis, divided according to disease activity into active (group A, n = 20) and non-active (group B, n = 18) patients. Pure tone audiometry, tympanometry and complete rheumatological assessment were performed. All patients presented poorer auditory thresholds compared with controls. Patients of group A had both air and bone conduction thresholds poorer than group B (although not statistically significant), and most patients of both groups presented an air-bone (a-b) gap. No significant difference in middle ear pressure was noticed between patients and controls. No correlation between hearing impairment and duration of the disease or patients' age was found. The high prevalence of hearing loss in autoimmune diseases supports the importance of audiometric evaluation in such patients. The auditory recovery through middle ear surgery before cranial nerve involvement could be considered in selected patients. Further investigations are needed for a better knowledge of the middle and inner ear involvement in patients with rheumatoid arthritis.     

周围神经脱髓鞘豚鼠模型听神经病变及听功能研究

目的:通过建立实验性变态反应性神经炎这一周围神经脱髓鞘动物模型,观察其听神经病变,初步探讨其听性脑干反应(ABR)和听神经复合动作电位(CAP)的改变。方法:以粗提的牛外周神经髓鞘碱性蛋白(MBP)作为抗原,免疫实验组豚鼠;对照组以生理盐水代替MBP。检测动物血清抗MBPIgG水平、坐骨神经传导速度,观察坐骨神经、听神经病理改变;检测ABR、CAP阈值及潜伏期;观察内耳病理损伤。结果:实验组血清抗MBPIgG水平升高,与对照组相比P<0.01;实验组坐骨神经传导速度减慢,与对照组相比P<0.05;透射电镜发现坐骨神经、听神经脱髓鞘改变;免疫前后实验组14只(26耳)出现ABR反应阈升高,伴Ⅰ、Ⅲ、Ⅴ波潜伏期明显延长,与对照组比较P<0.01,而Ⅰ～Ⅲ、Ⅲ～Ⅴ波间期与对照组比较P>0.05;CAPN1、N1波潜伏期延长,与对照组比较P<0.01;另有4只(8耳)仅出现潜伏期延长而无阈值升高;免疫组织化学显示内耳免疫损伤部位主要在蜗神经、内耳神经纤维、螺旋神经节;扫描电镜显示内毛细胞纤毛紊乱、胞质溢出。结论:实验性变态反应性神经炎动物模型作为一种可靠的周围神经脱髓鞘动物模型,其病变可累及听神经出现听神经脱髓鞘改变,ABR和CAP阈值升高、潜伏期明显延长,该模型可望成为探讨听神经脱髓鞘的听力学表现的一种有用的动物模型。     

A rat T-cell line that mediates autoimmune disease of the inner ear in the Lewis rat.

In various patterns of sensorineural hearing loss including Ménière's disease, which may show improvement in auditory function following immunosuppressive therapy, an isolated autoimmune disease of the inner ear has been postulated. Because of the lack of well-defined diagnostic criteria to identify autoimmune processes within the inner ear and the fact that the human inner ear is one of the few organs of the body not amenable to diagnostic biopsy, there has been great interest in developing animal models that mimic these clinical entities. Previous studies have found evidence that this process might be cell mediated and that the endolymphatic sac functions as an immunodefensive organ for the inner ear. By heterologous immunization of inbred Lewis rats with inner ear tissue, an autoreactive inner-ear-specific T helper cell line was established. After passive transfer of these cells a labyrinthitis was induced in recipient animals. Immunohistochemically, T helper cells were first identified in the cochlea suggesting that this cell type might carry the autoantigenic epitope. Autoantibodies against inner ear tissue were demonstrated in animals with histologically evident labyrinthitis. We conclude that this experimental design can serve as an animal model for cell-mediated autoimmune disease of the inner ear and could be used to explain the etiology of certain types of sensorineural hearing loss such as Ménière's disease. With this approach the identification of the causative autoantigen should be possible and will lead to the development of appropriate clinical tests to diagnose autoimmune diseases of the inner ear in humans.     

Frequency of cochlear enhancement on magnetic resonance imaging in patients with autoimmune sensorineural hearing loss

OBJECTIVE: To evaluate magnetic resonance imaging (MRI) scans for enhancement of inner ear structures of patients with sensorineural hearing loss and documented antibodies to the 68-kd inner ear antigen. STUDY DESIGN: Retrospective case review with reexamination of MRI scans. SETTING: Outpatient office. PATIENTS: Thirty-five patients with autoimmune sensorineural hearing loss defined by audiograms documenting a sensorineural hearing deficit in one or both ears and the presence of an anti-inner ear antibody (68-kd band) in serum samples who underwent precontrast and postcontrast T1-weighted axial and coronal MRI scans of the inner ear, which were performed concurrently with the hearing loss. INTERVENTIONS: Diagnostic. MAIN OUTCOME MEASURES: Frequency and intensity of cochlear enhancement on MRI scans. RESULTS: One patient demonstrated +2 cochlear enhancement. However, that finding was thought to represent postoperative inflammatory change. CONCLUSION: No correlation was found between the presence of antibodies to inner ear antigen in patients with hearing loss and cochlear enhancement on MRI scans.