蛋氨酸对离体培养小鼠耳蜗毛细胞顺铂耳毒性的拮抗作用

目的 在培养基中加入顺铂,建立离体培养小鼠耳蜗毛细胞的顺铂损害模型,探讨蛋氨酸对毛细胞的保护作用.方法 32只出生后2 d昆明小鼠,取出耳蜗基底膜,分为4组,每组16个样本,分别用无血清培养液、含顺铂的无血清培养液、含顺铂+蛋氨酸的无血清培养液以及含蛋氨酸的无血清培养液进行离体培养.采用肌球蛋白Ⅵ(myosinⅥ)免疫荧光染色,通过激光共聚焦显微镜、光镜、毛细胞计数等方法,观察蛋氨酸对顺铂引起的耳蜗毛细胞损伤的保护作用.结果 对照组和蛋氨酸组基底膜内、外毛细胞未见损伤缺失;顺铂组耳蜗基底膜内、外毛细胞均有不同程度的损伤,与对照组和蛋氨酸组比较,差异均具有统计学意义(P值均＜0.05);顺铂+蛋氨酸组基底膜内、外毛细胞的损伤程度与顺铂组相比明显减轻,差异具有有统计学意义(t内毛细胞=3.929、t外毛细胞=8.582,P值均＜0.05).结论 顺铂能损伤离体培养的小鼠耳蜗基底膜毛细胞,蛋氨酸可以在一定程度上拮抗顺铂对毛细胞的损伤.

Abstract：

Objective To establish an in vitro model of mouse cochlear basilar membrane impairment using cisplatin, and observe the protective effect of methionine on the hair cells. Methods The cochlear basilar membrane samples of thirty two Kunming mice were harvested on the 2nd day after birth and randomly divided into four groups. Each group had 16 samples. Overnight preincubation the cochlear organ followed by appropriate treatment respectively as follows: the serum-free culture medium, the serum-free culture medium with methionine and cisplatin, the cisplatinum-containing serum-free culture medium, and the methionine-containing serum-free culture medium. The protective effect of methionine for injury of cochlea hair cells induced by cisplatin was observed by myosin-Ⅵ immunofluorescence, lightmicroscop,laser confocal scanning microscope and hair cells counting. Results The outer hair cells(OHC) and inner hair cells(IHC) of control group and methionine group were not damaged. The outer and inner hair cells of cisplatin group were damaged in various degree, and had remarkable difference compared with control group and methionine group(P ＜ 0. 05). The outer hair cells and inner hair cells of cisplatin + methionine group were damaged less than the cisplatin group with remarkable difference (tIHC = 3. 929, tOHC = 8. 582, P ＜0. 05). Conclusions Cisplatinum could damage the cochlear hair cells of the basal membrane in Kunming mice. Methionine might protect against cisplatin's damage on the cochlear hair cells.

In vitro protective effect of methionine against cisplatin's damage to the cochlear hair cell of mice

Objective To establish an in vitro model of mouse cochlear basilar membrane impairment using cisplatin, and observe the protective effect of methionine on the hair cells. Methods The cochlear basilar membrane samples of thirty two Kunming mice were harvested on the 2nd day after birth and randomly divided into four groups. Each group had 16 samples. Overnight preincubation the cochlear organ followed by appropriate treatment respectively as follows: the serum-free culture medium, the serum-free culture medium with methionine and cisplatin, the cisplatinum-containing serum-free culture medium, and the methionine-containing serum-free culture medium. The protective effect of methionine for injury of cochlea hair cells induced by cisplatin was observed by myosin-Ⅵ immunofluorescence, lightmicroscop,laser confocal scanning microscope and hair cells counting. Results The outer hair cells(OHC) and inner hair cells(IHC) of control group and methionine group were not damaged. The outer and inner hair cells of cisplatin group were damaged in various degree, and had remarkable difference compared with control group and methionine group(P ＜ 0. 05). The outer hair cells and inner hair cells of cisplatin + methionine group were damaged less than the cisplatin group with remarkable difference (tIHC = 3. 929, tOHC = 8. 582, P ＜0. 05). Conclusions Cisplatinum could damage the cochlear hair cells of the basal membrane in Kunming mice. Methionine might protect against cisplatin's damage on the cochlear hair cells.