α-硫辛酸对顺铂致小鼠耳蜗毛细胞损伤的保护作用的体外研究

目的:研究α-硫辛酸(LA)对顺铂致离体小鼠耳蜗毛细胞损伤的保护作用。方法:分离3～5d的新生昆明种小鼠耳蜗基底膜,在含有1%牛血清白蛋白培养基中培养24h,分为对照组、模型组(顺铂16μg.mL-1)、药物对照(LA0.5mmol.L-1)组和低、高浓度LA(顺铂16μg.mL-1+LA0.2、0.5mmol.L-1)组(n=10),分别加入含相应药物的新鲜培养基2mL,培养24h,观察各组耳蜗毛细胞组织形态变化,计算内、外毛细胞缺失百分率,检测Bax、Bcl-2的表达情况。结果:与对照组比较,模型组小鼠耳蜗毛细胞出现纤毛倒伏、排列紊乱,内、外毛细胞缺失百分率、Bax表达水平明显增加(P<0.01),Bcl-2表达明显减弱(P<0.01);与模型组比较,低、高浓度LA组小鼠耳蜗毛细胞的形态改变明显减轻,内、外毛细胞缺失百分率、Bax表达明显减弱(P<0.05或P<0.01),Bcl-2表达明显增强(P<0.05),且高浓度组改善效果明显优于低浓度组(P<0.05);对照组与药物对照组比较无明显差异(P>0.05)。结论:LA对离体培养的顺铂致小鼠耳蜗毛细胞损伤具有保护作用。

Protective Effects of a-lipoic Acid against Cisplatin-induced Cochlea Hair Cells Injury in Mice in Vitro

OBJECTIVE：To investigate the protective effect of α-lipoic acid（LA） against cisplatin-induced cochlea hair cells impairment in mice in vitro.METHODS：The cochlea basilar membrane was isolated from postnatal day 3 to 5 Kunming mice cochlea and cultured in culture medium containing 1% bovine serum albumin for 24 hours.Those samples were divided into control group,model group（cisplatin 16 μg·mL-1）,drug control group（LA 0.5 mmol·L-1） and LA low and high concentration groups（cisplatin 16 μg·mL-1＋LA 0.2,0.5 mmol·L-1）（n=10）.They were cultured in 2 mL new culture medium containing relevant medicine for 24 h respectively.The morphology changes of mice cochlea hair cells were observed.The percentages of internal and external hair cell deficiency were calculated.At the same time,Bax and Bcl-2 expression in cochlea hair cell of mice were determined.RESULTS：Compared with control group,the cilia of cochlea hair cells in model group were lodged and arranged in disorder.The percentage of missing inner and outer hair cell and Bax expression were significantly increased（P〈0.01）,while Bcl-2 expression was decreased significantly（P〈0.01）;compared with model group,the morphological changes of cochlea hair cells in LA groups were obvious alleviated,and the percentage of missing hair cells and Bax expression were significantly decreased（P〈0.05 or P〈0.01）,while Bcl-2 expression was increased significantly（P〈0.05）,the improvement of high concentration group was better than low concentration group（P〈0.05）;there was no significant difference between control group and drug control group（P〉0.05）.CONCLUSION：This study demonstrates LA might effectively protect cochlea hair cells against cisplatin-induced injury in mice cultured in vitro.