CAJ | 학술논문

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.
목적 재배양기중가입순박,건립리체배양소서이와모세포적순박손해모형,탐토단안산대모세포적보호작용.방법 32지출생후2 d곤명소서,취출이와기저막,분위4조,매조16개양본,분별용무혈청배양액、함순박적무혈청배양액、함순박+단안산적무혈청배양액이급함단안산적무혈청배양액진행리체배양.채용기구단백Ⅵ(myosinⅥ)면역형광염색,통과격광공취초현미경、광경、모세포계수등방법,관찰단안산대순박인기적이와모세포손상적보호작용.결과 대조조화단안산조기저막내、외모세포미견손상결실;순박조이와기저막내、외모세포균유불동정도적손상,여대조조화단안산조비교,차이균구유통계학의의(P치균＜0.05);순박+단안산조기저막내、외모세포적손상정도여순박조상비명현감경,차이구유유통계학의의(t내모세포=3.929、t외모세포=8.582,P치균＜0.05).결론 순박능손상리체배양적소서이와기저막모세포,단안산가이재일정정도상길항순박대모세포적손상.
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.