缺氧对体外培养耳蜗螺旋神经节细胞及神经纤维的影响  

Effect of the spiral ganglion cell and nerve fiber of rat cochlea in vitro to hypoxia

作　　者：王丽萍[1] 王苹[2] 杜波[2] 杜宝东[2] 

机构地区：[1]吉林大学第一医院病理科,长春130021 [2]吉林大学第一医院耳鼻咽喉-头颈外科

出　　处：《临床耳鼻咽喉头颈外科杂志》2008年第22期1040-1042,共3页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

基　　金：世界卫生组织国际合作项目(No:2006.0781)

摘　　要：目的:建立耳蜗器官体外培养模型,详细观察缺氧对体外培养耳蜗螺旋神经节细胞(SGC)及神经纤维的影响。方法:分离生后3 d Wistar大鼠耳蜗基底膜,平铺在含有DMEM培养液的培养基内(每盘6条基底膜),2盘为1组,随机分为5组。4组作为实验组,按不同时间段(6 h、12 h、24 h、48 h)进行缺氧(37℃,90%N2,5%CO2,5%O2)培养。1组作为对照组放置在37℃、5%CO2培养箱。用抗神经丝蛋白作为一抗,对耳蜗SGC及神经纤维进行免疫荧光染色。激光共聚焦显微镜下观察缺氧时耳蜗SGC及神经纤维形态变化并计数单位面积(24 mm×36 mm)SGC数即细胞密度。结果:缺氧早期(6 h)神经纤维局灶性水肿,SGC变化不明显。12 h神经纤维局灶性断裂及崩解,SGC减少,细胞密度与对照组相比差异有显著性(P<0.01)。随缺氧时间延长,神经纤维崩解和SGC缺失逐渐加重,48 h神经纤维完全崩解破坏,SGC严重缺失,细胞密度与对照组比较差异有显著性(P<0.01)。结论:缺氧造成体外培养耳蜗SGC及神经纤维损伤,神经纤维对缺氧更敏感。Objective：To establish a practical model for Wistar rat cochlea organ culturing in vitro, and to ob serve the growing status in hypoxia of the spiral ganglion cell and nerve fiber. Method：We used an in vitro hypoxia model and dissociated cultures of the basal membrane from the cochlea of 3-day-old Wistar rats. And put them in incubator（37 ℃, 90 % N2, 5 % CO2, 5 % O2 ） to hypoxia culture for different times. The culture were Immunofiuo rescence dyed and count the number of the spiral ganglion ceil and the cell density in unit area（24 mm× 36 mm）, and observe the morph of nerve fiber under the confocal microscope, the results were compared with controls. Result：Hypoxia early （6 h） nerve fiber appear edema, spiral ganglion cell didn＇t change compared with controls; nerve fiber appear break and disintegration and the spiral ganglion cell decrease in 12 hours culturing, and the cell density in unit area had remarkable difference compared with control（ P 〈0.01）. Hypoxia leads to the cell density decrease in a time-dependent manner, the longer of cultures times in hypoxia, the heavier of damage in spiral ganglion cell and nerve fiber. Twelve hours culturing, and the cell density in unit area had remarkable difference compared with control（ P 〈0.01）. Hypoxia leads to the cell density decrease in a time-dependent manner, the longer of cultures times in hypoxia, the heavier of damage in spiral ganglion cell and nerve fiber. Conclusion： The study findings suggest that hypoxia makes the spiral ganglion cell and nerve fiber damage of culturing in vitro, and nerve fiber more susceptible than spiral ganglion cell for hypoxia.

关 键 词：耳蜗 螺旋神经节细胞 神经纤维 体外培养 缺氧 

分 类 号：R764.35[医药卫生—耳鼻咽喉科]