不同浓度Ouabain对贵州小型猪耳蜗形态及听功能影响的初步研究

发布时间：2018-02-03 14:04

本文关键词： Ouabain 毛细胞螺旋神经节听力损失　出处：《南京医科大学》2013年硕士论文　论文类型：学位论文

【摘要】：Ouabain是一种有效的Na+/K+-ATP酶抑制剂，曾作为临床治疗心衰的药物使用，其地位现已经被地高辛所取代。在内耳中，同样分布着大量的Na+/K+-ATP酶，其对维持内、外淋巴钾离子梯度及各细胞功能稳定起着重要作用。研究显示，将Ouabain应用于动物内耳，可造成不同程度的螺旋神经节细胞和毛细胞损害。在沙鼠，Ouabain可特异性损伤螺旋神经节I型神经元，制造出听神经病动物模型，进一步用于听神经病发病机制及治疗方法的研究；而在豚鼠，ouabain主要损伤耳蜗毛细胞。贵州小型猪与人类的种属差异性更小，在很多疾病与人类有共同的发病机制，，在内耳疾病研究中，小型猪与啮齿类动物相比也具有很多优势。本实验中选用在进化上与人类更为接近的小型猪，进一步研究ouabain对大体型动物内耳的影响。目的：探讨短时间局部应用不同浓度ouabain溶液对贵州小型猪内耳的作用，分析内耳形态及听力改变与药物浓度关系。方法：于圆窗龛处局部应用不同浓度ouabain或生理盐水，实验动物分为1mMouabain组、0.1mMouabain组、生理盐水对照组及空白对照组。采用听性脑干反应（auditorybrainstemresponse，ABR）测定、免疫组织化学染色等方法，观察各组动物在应用药物后3天、一周及两周时耳蜗内螺旋神经节和毛细胞的形态、数目及听功能变化。结果：生理盐水对照组及空白对照组在术后内耳形态及听功能方面无明显变化。与空白对照组相比，1mM组螺旋神经节细胞明显减少，0.1mM组损伤相对较轻；术后两周时，1mM组毛细胞排列紊乱，数量无明显减少，0.1mM组毛细胞基本无变化；1mM组及0.1mM组ABR阈值均较对照组显著升高（P0.01），其中1mM组比0.1mM组升高更明显（P0.05）。结论：不同浓度ouabain溶液对贵州小型猪螺旋神经节细胞均有损害作用，损伤程度与药物浓度呈正相关，耳蜗毛细胞受损相对较轻，而对听功能的影响较为明显。
[Abstract]:Ouabain is an effective Na / K -ATPase inhibitor, which has been used as a clinical drug for the treatment of heart failure, and its status has been replaced by digoxin. There are also a large number of Na / K-ATPase, which plays an important role in maintaining the gradient of potassium ions in the endolymphatic and perilymphatic and the functional stability of the cells. It has been shown that Ouabain is used in the inner ear of animals. In the gerbils, the spiral ganglion type I neurons were specifically damaged, and the animal model of auditory neuropathy was made. To further study the pathogenesis and treatment of auditory neuropathy; However, in guinea pigs, the cochlear hair cells were mainly damaged. The difference between Guizhou miniature pigs and human species was even smaller, and there were common pathogenesis between many diseases and human beings, and in the study of inner ear diseases. Compared with rodents, miniature pigs have many advantages. In this study, the effects of ouabain on the inner ear of large animals were studied. Objective: to investigate the effects of local administration of different concentrations of ouabain solution on the inner ear of Guizhou miniature pig, and to analyze the relationship between the shape of the inner ear and the changes of hearing and drug concentration. Methods: the animals were divided into 1 m Mouabain group and 0.1 m Mouabain group with different concentrations of ouabain or normal saline. The auditory brainstem response (ABR) and immunohistochemical staining were used in normal saline control group and blank control group. The morphology, number and auditory function of spiral ganglion and hair cells in cochlea were observed 3 days, 1 week and 2 weeks after administration. Results: there were no significant changes in the shape and auditory function of the inner ear in the saline control group and the blank control group, but the number of spiral ganglion cells decreased significantly in the 1 mm group compared with the blank control group. The injury was relatively mild in 0.1 mm group. At two weeks after operation, the hair cells in the 1st mm group were disordered, and the number of hair cells in 0.1 mm group was not significantly decreased. The ABR threshold of 1mm group and 0.1 mm group were significantly higher than that of the control group, especially the 1mm group was significantly higher than the 0.1 mm group. Conclusion: different concentrations of ouabain solution can damage spiral ganglion cells of Guizhou miniature pigs. The degree of injury is positively correlated with the concentration of drug, and the damage of cochlear hair cells is relatively mild. But the effect on hearing function is obvious.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R764.4;R965

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