中耳加压对豚鼠膜迷路积水及内耳AQP1、2表达的影响

发布时间：2018-04-05 02:13

  本文选题：水通道蛋白　切入点：醛固酮　出处：《辽宁医学院》2012年硕士论文

【摘要】：目的 制备豚鼠膜迷路积水动物模型,并对动物进行中耳加压治疗，观察中耳加压后膜迷路积水的变化及AQP1和AQP2表达的变化，以探究AQPs在积水过程中的可能作用及中耳加压对膜迷路积水影响的可能机制。 方法 采用腹腔注射醛固酮方法制备豚鼠膜迷路积水动物模型，将50只豚鼠随机分为5组，每组各10只，对照组及加压对照组生理盐水腹腔注射0.5ml/d，连续5d；积水4周组，积水8周组，加压组第1周腹腔注射醛固酮0.1mg/(100g/d),连续5d。加压组及加压对照组豚鼠在造模后第5周起置于压力舱内加压治疗，共3周，观察每组动物耳蜗形态和功能的变化，并用SP法免疫组织化学及Western blot方法检测各组豚鼠内耳AQP1、AQP2的表达变化。 结果 1、积水8周组ABR听阈值为[47.50±4.25]dBSPL比积水4周组[36.50±4.74]dBSPL、加压对照组[24.95±2.35] dBSPL和对照组[23.52±2.41] dBSPL升高(P0.05)，加压组[33.50±2.42] dBSPL较积水8周组阈值下降(P0.05)。 2、膜迷路积水程度：对照组及加压对照组无膜迷路积水；积水8周组R值为[0.63±0.11]较积水4周组[0.47±0.08]及加压组[0.42±0.05]升高（P0.05）。 3、免疫组化实验结果：①AQP1表达主要在螺旋韧带基底部、Corti’s器基底膜等处，积水8周组表达面积比值为[0.0946±0.0125]较积水4周组[0.1163±0.0148]、对照组[0.1492±0.0125]豚鼠AQP1的表达减少（P0.05），加压组[0.1216±0.0118]与积水8周组比较AQP1的表达增加（P0.05）。②AQP2表达主要在血管纹、螺旋韧带、Corti’s器等处，积水8周组表达面积比值为[0.2538±0.0157]较积水4周组[0.1743±0.0064]、对照组[0.1191±0.0085]表达增加（P0.05），加压组[0.1502±0.0112]与积水8周组比较AQP2的表达减少（P0.05）。 4、Western blot实验结果：①耳蜗外侧壁组织AQP1表达量在积水8周组[0.674±0.25]较积水4周组[0.856±0.12]和对照组[0.978±0.15]减少（P0.05），，加压组[0.753±0.32]与积水8周组比较表达量增加（P0.05）。②耳蜗外侧壁组织AQP2表达量在积水8周组[3.507±0.89]较积水4周组[2.194±0.75]及对照组[0.985±0.15]增加（P0.05），加压组[1.891±0.31]与积水8周组比较表达量减少（P0.05）。 结论 1、中耳加压可减轻醛固酮引起的豚鼠膜迷路积水，并且改善了耳蜗听功能。 2、由腹腔注射醛固酮引起豚鼠膜迷路积水时，在实验周期内积水随时间延长而加重，AQP1表达下调，而AQP2表达增加。 3、中耳加压治疗使膜迷路积水豚鼠耳蜗AQP1表达增加，AQP2表达减少，说明中耳加压减轻膜迷路积水可能与AQP的表达改变有关。
[Abstract]:PurposeThe animal model of membranous labyrinth hydrops in guinea pigs was established and treated with middle ear pressure. The changes of membranous labyrinthine hydrops and the expression of AQP1 and AQP2 were observed.To explore the possible role of AQPs in hydrops and the possible mechanism of the effects of middle ear pressure on membranous labyrinthine hydrops.Method50 guinea pigs were randomly divided into 5 groups by intraperitoneal injection of aldosterone. The control group and the pressurized control group were injected with 0.5 ml / d normal saline for 5 consecutive days, and the hydrops for 4 weeks were treated with hydrocephalus for 8 weeks, the control group and the pressurized control group were injected intraperitoneally with 0.5 ml / d of normal saline for 5 days.In the first week, aldosterone was injected intraperitoneally with 0.1 mg / 100 g / d aldosterone for 5 days.The guinea pigs in the compression group and the control group were placed in the pressure chamber for 3 weeks from the 5th week after the establishment of the model. The changes of cochlea morphology and function in each group were observed.The expression of AQP2 in the inner ear of guinea pigs was detected by SP immunohistochemical method and Western blot method.Result1,绉�姘�8鍛ㄧ粍ABR鍚�闃堝�间负[47.50卤4.25]dBSPL姣旂Н姘

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