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背景:一氧化氮由一氧化氮合酶催化而成,关于一氧化氮合酶的3种异构体在豚鼠耳蜗结构中的确切定位和表达,目前尚存争议。目的:观察一氧化氮合酶3种异构体在豚鼠耳蜗的定位表达情况,探讨一氧化氮在内耳听觉生理和病理生理过程中的作用。设计:观察对比实验。单位:锦州医学院生理学教研室、锦州市附属第二医院骨科及锦州市中心医院耳鼻咽喉科。材料:实验于2000-05/11在中国医科大学听力研究室完成。选用健康纯白红目雄性豚鼠10只,体质量250～300g,耳郭反射灵敏。方法:豚鼠经腹腔麻醉后,断头处死,速取听泡。打开圆窗和卵圆窗并缓慢灌流含40g/L多聚甲醛的磷酸盐缓冲液,再将标本浸入该溶液固定2h。100g/L乙二胺四乙酸溶液脱钙1周后,移入250g/L蔗糖溶液中过夜沉淀。包埋耳蜗,制成20μm恒冷冰冻切片用于免疫组织化学染色。主要观察指标:豚鼠耳蜗内一氧化氮合酶3种异构体的定位表达情况。结果:10只豚鼠全部进入结果分析,无脱失。①神经元型和内皮型一氧化氮合酶免疫反应阳性见于耳蜗螺旋器的内、外毛细胞和螺旋神经节细胞。此外,也见于血管纹边缘细胞、螺旋韧带细胞和神经纤维。②诱导型一氧化氮合酶免疫反应活性在生理状态下也见于耳蜗的上述各部位。结论:一氧化氮在哺乳类动物耳蜗神经传导、血流调节及细胞毒性等生理和病理生理过程中可能起重要作用。
BACKGROUND: Nitric oxide is catalyzed by nitric oxide synthase. The exact location and expression of three isoforms of nitric oxide synthase in the guinea pig cochlear structure remains controversial. OBJECTIVE: To observe the localization of nitric oxide synthase (NOS) in the cochlea of guinea pigs and to explore the role of nitric oxide in the auditory physiology and pathophysiology of the inner ear. Design: observe the contrast experiment. SETTING: Department of Physiology, Jinzhou Medical College, Department of Orthopedics, Second Affiliated Hospital of Jinzhou City, and Department of Otorhinolaryngology, Jinzhou Central Hospital. Materials: The experiment was performed at Hearing Laboratory, China Medical University from May to May 2000. Selection of healthy pure white male red guinea pigs 10, body weight 250 ~ 300g, sensitive to ear reflex. Methods: After intraperitoneal anesthesia in guinea pigs, the decapitated rats were sacrificed. Open the round window and oval window and slowly perfused phosphate buffer containing 40g / L paraformaldehyde, then the specimen was immersed in the solution for 2h. 100g / L ethylenediaminetetraacetic acid solution decalcification for 1 week, transferred to 250g / L sucrose solution overnight precipitation. Cochlear implants were used to make 20 [mu] m frozen frozen sections for immunohistochemical staining. MAIN OUTCOME MEASURES: Localization and expression of three isoforms of nitric oxide synthase in guinea pig cochlea. Results: All the 10 guinea pigs entered the result analysis without loss. ① Neuronal and endothelial nitric oxide synthase immunoreactivity positive in the cochlear spiral inner and outer hair cells and spiral ganglion cells. In addition, also found in stria vascularis cells, spiral ligament cells and nerve fibers. ② inducible nitric oxide synthase immunoreactivity in the physiological state is also seen in the above parts of the cochlea. Conclusion: Nitric oxide may play an important role in the physiological and pathophysiological processes of cochlear nerve conduction, blood flow regulation and cytotoxicity in mammals.