内皮舒张因子对豚鼠耳蜗血管纹血管的保护作用

本实验利用活体显微镜摄像技术、透射电镜技术,观察了内皮舒张因子（ＥＤＲＦ）对豚鼠耳蜗微循环的保护作用。结果提示：（１）速尿组（Ｆ）动物,经静脉注射药１０ｍｉｎ后,耳蜗微动脉缺血,血管内皮细胞损伤;（２）地速尿／Ｌ－精氨酸组（Ｆ／Ｌ－Ａｒｇｉｎｉｎｅ）动物,ＥＤＲＦ能使速尿引起的微动脉缺血明显改善,血管纹血管超微结构的损伤程度较单纯Ｆ组减轻;（３）速尿／Ｌ－硝基－精且（Ｆ／Ｌ－ＮＮＡ）动物,耳蜗的缺     

内皮舒张因子对耳蜗微循环作用的实验研究

为探讨内皮依赖性舒张因子（EDRF）对耳蜗微循环的调节作用，本实验应用活体显微镜结合电视摄象的图像分析技术及血管纹螺旋韧带铺片技术，观察了不同剂量EDRF合成前体L-精氨酸（L－Arg）及EDRF的合成抑制剂L-硝基-精氨酸（L－NNA）对耳蜗血流（cochlearbloodfrow，CoBF）的作用。结果显示：L－Arg100mg／kg组动物在给药5分钟后耳蜗血管流速开始加快，20分钟时平均增加达31．4％，血管扩张，平均30分钟恢复正常；L－Arg200mg／kg组动物，在给药早期耳蜗血管纹血流速度增加，之后流速减慢，20分钟时平均下降54％，并伴BP下降；而L－NNA组给药后血流速度减慢，血管收缩；同时对EDRF增加CoBF的机理进行了探讨。提示：①、EDRF对CoBF有调节作用；②适当剂量的EDRF可扩张血管，加快血流，增加耳蜗微循环的灌注量。     

9370兆赫微波与耳蜗微循环的研究

利用激光多普勒测试技术结合螺旋韧带血管纹红细胞计数，探讨微波对豚鼠耳蜗微循环的作用。结果表明：（１）微波组动物经辐射１０分钟后，耳蜗微循环的血流量较辐射前上升１９．７９％，３０分钟后上升２３．４５％；（２）对速尿／微波一组动物，微波能使速尿引起的血流下降明显改善。与此同时血管纹毛细血管的红细胞充盈也较单纯注射速尿动物组显著增加；（３）有关听功能的改善情况，比较ＡＰ反应阈恢复正常的时间，速尿／微波组动物可较单纯速尿组动物提前１／３。提示加快内耳微循环的血流，增加血管纹红细胞的充盈是微波辐射促进听功能恢复有重要作用。     

9370兆赫微波与耳蜗微循环的研究

利用激光多普勒测试技术结合螺旋韧带血管纹红细胞计数，探讨微波对豚鼠耳蜗微循环的作用。结果表明：（1）微波组动物经辐射10分钟后，耳蜗微循环的血流量较辐射前上升19．79%，30分钟后上升23．45%；(2)对速尿／微波-组动物，微波能使速尿引起的血流下降明显改善。与此同时血管纹毛细血管的红细胞充盈也较单纯注射速尿动物组显著增加；(3)有关听功能的改善情况，比较AP反应阈恢复正常的时间，速尿／微波组动物可较单纯速尿组动物提前1/3．提示加快内耳微循环的血流，增加血管纹红细胞的充盈是微波辐射促进听功能恢复有重要作用。     

听力学

980368内耳器官培养技术及其应用(综述)/崔鹏程…//陕西医学杂志一1997,26(5)一287一288980369内皮舒张因子对耳蜗微循环作用的实验研究/史晓瑞…//耳鼻咽喉一头颈外科一1 997,4(3)一172~176 为探讨内皮依赖性舒张因子(EDRF)对耳蜗微循环的调节作用,本实验应用活体显微镜结合电视摄象的图像分析技术及血管纹螺旋韧带铺片技术,观察了不同剂量EDRF合成前体L一精氨酸(L一Arg)及EDRF的合成抑制剂L-硝基一精氨酸(L一NNA)对耳蜗血流(C。BF)的作用。结果显示:I一Arg 100mg/kg组动物在给药5分钟后耳蜗血管流速开始加快,20分钟时平均增加达…     

缺血后处理减轻缺血再灌注大鼠耳蜗血管损伤的研究

目的探讨缺血后处理（ischemic postconditioning）对大鼠耳蜗缺血再灌注的保护作用。方法成年雄性SD大鼠42只,随机分为3组（n=14）,分别为假手术对照组、缺血再灌注组和缺血后处理组。各组动物于再灌注24h取左侧耳蜗,每组随机4个耳蜗做透射电镜观察血管纹血管的超微结构变化,各组余下耳蜗组织进行匀浆,测定匀浆中过氧化氢酶（catalas,CAT）活性、丙二醛（molondialdehyde,MDA）含量。结果与假手术对照组大鼠相比,缺血再灌注组和缺血后处理组大鼠CAT活性显著降低（P〈0.05）,MDA含量显著升高（P〈0.05）;与缺血再灌注组大鼠相比,缺血后处理组大鼠CAT活性升高（P〈0.05）,MDA含量下降（P〈0.05）。形态学缺血再灌注组耳蜗血管内皮细胞超微结构破坏明显,而缺血后处理组超微结构有明显改善。结论耳蜗缺血后处理可能抑制耳蜗缺血再灌注后的氧化损伤,对血管有一定保护作用。     

内皮舒张因子对耳蜗微循环作用的实验研究

为探讨内皮依赖性舒张因子（ＥＤＲＦ）对耳蜗微循环的调节作用，本实验应用活体显微镜结合电视摄象的图像分析技术及血管纹螺旋韧带铺片技术，观察了不同剂量ＥＤＲＦ合成前体Ｌ－精氨酸（Ｌ－Ａｒｇ）及ＥＤＲＦ的合成制抑制Ｌ－硝基－精氨酸（Ｌ－ＮＮＡ）对耳蜗血清（ｃｏｃｈｌｅａｒｂｌｏｏｄｆｒｏｗ，ＣｏＢＦ）的作用，结果显示：Ｌ－Ａｒｇ１００ｍｇ／ｋｇ组药物在给药５分钟后蜗血管流速开始加快，２０分钟时平均增加达     

豚鼠耳蜗微动脉自律运动的连续观察及分析

目的探讨耳蜗微动脉自律运动的频率特性、收缩特性的特征参数。方法采用微循环数字图像检测和分析方法，根据微动脉管径动态变化的跟踪测量结果，利用快速富里叶频谱转换进行定量分析。结果正常状态下耳蜗微动脉存在运动幅度较小的自律运动，自律运动的频率为（０．０２１±０．００２）Ｈｚ，振幅为（０．３７±０．１１）μｍ，管径变化范围为（１２．２９±１．９０）μｍ～（１０．４１±３．０３）μｍ，平均管径为（１１．２０±２．７０）μｍ，收缩指数为０．４２±０．１７。静脉注入Ｌ精氨酸后，耳蜗微动脉的血管运动被激活，在给药１０ｍｉｎ时振幅达（０．８４±０．０１）μｍ，频率增加为（０．０６８±０．００２）Ｈｚ，与用药前相比有显著的统计学意义。结论正常耳蜗微动脉存在运动幅度较小的自律运动，Ｌ精氨酸能显著增加微动脉自律运动的频率及振幅，增强了耳蜗微循环的功能，使耳蜗血流量增加。     

速尿引起的小鼠耳蜗血管纹缺血缺氧性病变

目的观察腹腔注射速尿引起的小鼠耳蜗血管纹缺血缺氧病变过程,探讨速尿破坏蜗管外壁开放血-迷路屏障的最佳时机。方法选择42只听力正常的健康CBA小鼠用于本实验。其中24只小鼠被平均分为3组,全麻下分别按照每公斤体重400毫克或200毫克或100毫克的剂量腹腔注射速尿,同时监测注射速尿前后的听性脑干反应的变化,在用药后1周终止实验并记录各组动物的存活数量,以确定不同剂量速尿对听觉功能的影响及其安全性。另外18只小鼠被平均分为6组,分别在注射速尿前和注射速尿后10分钟,30分钟,1小时,3小时和6小时随机处死3只小鼠,用于观察腹腔注射速尿后引起的血管纹缺血缺氧性病理改变过程。受试动物的一侧耳蜗制作成全耳蜗螺旋韧带铺片并用伊红染色,观察血管纹毛细血管内的红血球充盈情况以评估血管纹的供血在注射速尿后不同时间的缺血程度,另一侧耳蜗用环氧树脂法包埋后切片,观察血管纹上皮细胞因组织缺血缺氧而引起的病理学改变。结果按照每公斤体重400毫克的剂量腹腔注射速尿引起的小鼠死亡率约为75%,按照每公斤体重100毫克剂量腹腔注射速尿引起的小鼠听性脑干反应仅发生短时间的中等程度阈移,而按照每公斤体重200毫克剂量腹腔注射速尿的小鼠死亡率小于15%,但可造成较长时间的严重耳聋,因此按照每公斤体重200毫克的剂量腹腔注射速尿是实现小鼠暂时性耳聋的安全有效剂量。耳蜗螺旋韧带铺片结果显示按照每公斤体重200毫克的剂量腹腔速尿注射后10分钟即开始发生供血减少;用药后30分钟和1小时,血管纹呈现许多空血管;用药后3小时,毛细血管内的红细胞数量开始有所增加;但直到用药后6小时仍未完全恢复。血管纹切片结果显示,用药后10分钟,血管纹上皮细胞已经开始因缺氧而发生肿胀;用药后30分钟和1小时,血管纹细胞内出现许多空泡;用药后3小时和6小时,血管纹上皮细胞的积水肿胀仍然十分严重。结论腹腔注射速尿后可在数分钟内阻断供应耳蜗外侧壁的血流,血管纹的供血在用药后6小时逐渐趋向于恢复,但因缺血引起的血管纹细胞缺氧性肿胀和破坏还可持续一段更长的时间。从腹腔注射速尿后1小时至少到6小时可能都是蜗管外壁血-迷路屏障因缺氧变性而暂时性开放的重要时机。     

耳蜗外侧壁微区微循环损伤对耳蜗内淋巴电位的影响

为了解某些内耳疾病引起的感音神经性聋的区域频率听力下降与耳蜗局部血流改变的关系，采用光化学方法造成耳蜗外侧壁微区微循环血管损伤，应用活体耳蜗微循环与内淋巴电位（ＥＰ）同时观察记录的方法，观察了不同损伤面积的微循环改变对内淋巴电位的影响。实验应用血卟啉单甲醚作为光敏剂，超高压汞灯作为激发光。实验发现相对较小面积的耳蜗外侧壁微循环损伤组（０．２ｍｍ×０．４ｍｍ）ＥＰ无明显变化；相对较大面积的损伤组（０．２ｍｍ×０．８ｍｍ）ＥＰ轻度下降，但这种下降与正常对照组ＥＰ变化相比差异无显著性。结果提示ＥＰ虽产生于血管纹，但小面积血管纹微循环损伤不引起ＥＰ的明显变化，说明耳蜗局部代偿能力能够在一定程度上维持内耳功能。     

The histologic study of the inner ear in guinea pigs on anaphylatoxin

Complement is known to relate to many inflammatory reactions. C4a, C3a and C5a, known as anaphylatoxins, are known to cause strong inflammatory reactions. In this study, the role of anaphylatoxins on the pathogenesis in the cochlea was examined. On hundred forty six male Harley guinea pigs, weighing about 350 grs, all susceptible to preyer's reflex, were used in this study. Anaphylatoxins were made from guinea pig serum treated with zymosan, and inoculated into the carotid artery of the guinea pigs. Parts of these animals were sacrificed and examined at ten minutes, one day, two days, three days, seven days, ten days and fifteen days after injection of anaphylatoxins. Pathological changes in inner ears were observed by light microscopy. After 10 minutes, inner ears were found morphologically normal. After one day, inner ears were found to be almost morphologically normal but the stria vascularis was observed with cystic formation. After two days, cystic formations in the stria vascularis were enlarged and Reissner's membranes were collapsed in some other animals. After three days, the stria vascularis in the various cochlear turns except in the basal turn, were extremely atrophied, some cochlear nerves showed degeneration and some cochlea showed endolymphatic hydrops. After seven days, ten days and fifteen days, the morphological changes showed atrophy in the stria vascularis similar to the results observed on the third days. Atrophy in the stria vascularis was improved gradually with time, but the degeneration of the cochlear nerve was not improved. Opinions have been divided on the cause of inner ear disease including Meniere's disease. Many authors have reported that infectious diseases, for example mumps, measles and cytomegalovirus infection, have caused human sensorineural hearing loss. These diseases have been reported to result in atrophy in the stria vascularis, degeneration of the cochlear nerve and some other pathological changes. In this study, it was clearly observed that the atrophy of the stria vascularis, the endolymphatic hydrops and other morphological changes were caused by introduction of anaphylatoxins. These results were similar to the pathological changes observed in inner ear diseases in human beings. Therefore, inflammatory substances, including anaphylatoxins, were closely related to the cause of inner ear diseases. The animal model used in this report is considered to be important for elucidating the pathogenesis of inner ear diseases.     

内皮素在噪声性内耳损伤过程中的作用

目的研究噪声性内耳损伤过程中耳蜗微循环的病理变化以及内皮素(endothelin，ET)在这一病理过程中的作用及意义。方法30只大鼠按电脑产生随机数字法分为5组：对照组、噪声暴露(115dB白噪声每天8h)1、4、8和15d组。分别用扫描电镜观察毛细胞的形态，血管纹铺片观察耳蜗微循环的状态，放射免疫方法检测血浆ET含量，免疫组化染色观察ET-1、ETA、ETB的分布。结果噪声4、8、15d组出现了血管纹明显缺血，外毛细胞形态异常。血浆ET于4d组出现暂时性升高。ET-1于耳蜗组织中有广泛的阳性表达，噪声暴露前后差异无统计学意义；ETA主要分布于血管纹中间细胞与微血管壁周围的细胞，对照组与1d组表达呈弱阳性( )，噪声4、8、15d组表达呈强阳性( )。ETB主要分布于血管纹毛细血管内皮细胞，其变化趋势与ETA相似。结论噪声性内耳损伤过程中出现了明显的耳蜗微循环障碍，同时大鼠耳蜗组织的ET系统活性增高，二者在时间上有明显的一致性。推测噪声所致的耳蜗微循环障碍过程中ET起了重要的介导作用。     

In vivo imaging of mammalian cochlear blood flow using fluorescence microendoscopy.

AIMS: We sought to develop techniques for visualizing cochlear blood flow in live mammalian subjects using fluorescence microendoscopy. BACKGROUND: Inner ear microcirculation appears to be intimately involved in cochlear function. Blood velocity measurements suggest that intense sounds can alter cochlear blood flow. Disruption of cochlear blood flow may be a significant cause of hearing impairment, including sudden sensorineural hearing loss. However, inability to image cochlear blood flow in a nondestructive manner has limited investigation of the role of inner ear microcirculation in hearing function. Present techniques for imaging cochlear microcirculation using intravital light microscopy involve extensive perturbations to cochlear structure, precluding application in human patients. The few previous endoscopy studies of the cochlea have suffered from optical resolution insufficient for visualizing cochlear microvasculature. Fluorescence microendoscopy is an emerging minimally invasive imaging modality that provides micron-scale resolution in tissues inaccessible to light microscopy. In this article, we describe the use of fluorescence microendoscopy in live guinea pigs to image capillary blood flow and movements of individual red blood cells within the basal turn of the cochlea. METHODS: We anesthetized eight adult guinea pigs and accessed the inner ear through the mastoid bulla. After intravenous injection of fluorescein dye, we made a limited cochleostomy and introduced a compound doublet gradient refractive index endoscope probe 1 mm in diameter into the inner ear. We then imaged cochlear blood flow within individual vessels in an epifluorescence configuration using one-photon fluorescence microendoscopy. RESULTS: We observed single red blood cells passing through individual capillaries in several cochlear structures, including the round window membrane, spiral ligament, osseous spiral lamina, and basilar membrane. Blood flow velocities within inner ear capillaries varied widely, with observed speeds reaching up to approximately 500 microm/s. CONCLUSION: Fluorescence microendoscopy permits visualization of cochlear microcirculation with micron-scale optical resolution and determination of blood flow velocities through analysis of video sequences.     

内耳色素对爆震性听损伤保护作用的实验观察

目的　探讨内耳色素与爆震性听损伤的关系。方法　利用耳蜗铺片、石蜡切片、透射电镜及扫描电镜观察爆震前后白化豚鼠和杂色豚鼠耳蜗形态结构的变化 ,并对爆震前后白化豚鼠和杂色豚鼠听性脑干反应(ABR)反应阈进行测定。结果　白化豚鼠的耳蜗形态损伤和听功能损伤均较杂色豚鼠严重。结论　爆震后白化豚鼠耳蜗形态学损伤及ABR反应阈的改变均较杂色豚鼠明显。提示内耳血管纹色素颗粒与爆震性内耳损伤有关 ,机理可能为色素颗粒参与调节爆震后内淋巴液中钙离子浓度的平衡及参与清除爆震后耳蜗产生的氧自由基     

Effects of indomethacin in guinea pig cochlea: researches on audiology, morphology and cochlear blood flow]

Prostaglandins are known to be widely distributed in mammal tissues with diverse and potent biological activities. Indomethacin is a well-known cyclooxygenase inhibitor of prostaglandin synthesis. However, no paper appeared in the literature with regard to the effects of indomethacin on inner ear hearing function and cochlear lateral wall blood flow (LWBF). The present project was undertaken to investigate the effect of super-tolerated doses of indomethacin on audiology, morphology and LWBF of cochlea when that endogenous prostaglandin synthesis was greatly suppressed. In drug-treated group, super-tolerated doses of indomethacin (100 mg/kg) were given iv. to 29 guinea pigs. The compound action potentials (CAP) were monitored before and after drug administration till 180 minutes. The morphological changes of the organ of Corti were examined with scanning electron microscope, and that of stria vascularis with light and transmission electron microscopes. The LWBF were measured by biomicrosphere technique. The data were compared to those of the control group. Results showed no statistically significant changes of amplitude and threshold of CAP after indomethacin injection. No obvious morphological damage was found in the organ of Corti and stria vascularis except that a few of myelin figure were observed in marginal cell. LWBF was not decreased at 30 and 120 minutes after indomethacin injection. The study suggests that therapeutic doses of indomethacin do not damage cochlea, and inner ear hearing function is not severely interfered with when endogenous PGs synthesis is greatly suppressed, at least over the short time.     

Distribution of beta-tubulin in guinea pig inner ear

Our previous research had suggested that beta-tubulin might be an autoantigen for autoimmune inner ear disease. In this study, the expression of beta-tubulin in inner ears of normal and tubulin-immunized guinea pigs was examined by immunohistochemical staining. Strong immunoreactivity to beta-tubulin monoclonal antibody was found in stria vascularis, neurons of the spiral ganglion, cochlear nerve fibers and spiral ligament. Diffuse staining was found in the stria vascularis and the neurons of the spiral ganglion, while dense network staining was found in the spiral ligament, the nerve fibers and the vestibular end organs. The semicircular canals, endolymphatic duct and sac were also positively stained. In inner ears of guinea pigs challenged with beta-tubulin, staining intensity was diminished in the stria vascularis, the spiral ligament, and the neurons of the spiral ganglion. The results suggest that beta-tubulin is distributed to most structures of guinea pig inner ear. A challenge to the inner ear by tubulin could change the beta-tubulin distribution and cause degeneration in the spiral ganglion. The results support the hypothesis that beta-tubulin might be an autoantigen for autoimmune inner ear disease.     

噪声刺激对耳蜗一氧化氮合酶的影响

目的：探讨一氧化氮（ＮＯ）在噪声性聋发病中的作用。方法：用中高频连续稳态噪声制作噪声性聋的动物模型,用ＮＡＤＰＨ－黄递酶组织化学、原位杂效和Ｎｏｒｔｈｅｒｎ印迹法,观察噪声刺激对耳蜗一氧化氮合酶（ＮＯＳ）表达的影响。结果：组织化学法显示ＮＯＳ主要分布于内外毛细胞、螺旋神经节细胞和血管纹边缘细胞;原位杂效法发现ＮＯＳｍＲＮＡ在内外毛细胞、螺旋神经节细胞胞浆内均可见阳性染色,但血管纹边缘细胞无阳性染色     

Experimental labyrinthitis in guinea pigs caused by a hantavirus

Hantavirus-induced acute viral infection of the inner ear of guinea pigs is reported. Guinea pigs were inoculated directly into the inner ear through the round window, or intraperitoneally with hantavirus (B-1 strain), and the cochlea was examined using an immunohistochemical technique for the localization of viral antigens. Virus-specific antigens were detected prominently in the stria vascularis and to a lesser degree in other inner ear structures of the infected guinea pigs. The morphological changes in the inner ear structures of these guinea pigs were observed. Virus was isolated from the blood of some infected guinea pigs. These data suggest that hantavirus can infect the inner ear of the guinea pig via viremia. The effect of immunosuppression by ciclosporin A or cyclophosphamide is discussed.     

iNOS在链霉素耳中毒豚鼠耳蜗血管纹的表达

目的探讨链霉素（streptomycin,SM）耳中毒过程中豚鼠耳蜗诱生型一氧化氮合霉（inducible nitric oxide synthase,iNOS）的表达.方法 20只豚鼠随机分为正常对照组、SM组,每组10只,SM组每日腹腔注射硫酸链霉素150 mg/kg,正常对照组每日腹腔注射等量生理盐水,用药10天后处死.以听性脑干反应（ABR）为监测指标,结合电镜、免疫组化及图像分析技术,检测链霉素耳中毒过程中耳蜗血管纹iNOS的表达及形态学改变.结果用药10天后,SM组ABR阈值明显高于正常对照组,有统计学差异（P〈0.01）,电镜下可见SM组血管纹损伤严重;免疫组化结果表明,SM组血管纹iNOS的表达明显高于正常对照组.结论 SM耳中毒时ABR阈值升高,血管纹iNOS表达增强.     

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