顺铂对离体培养小鼠耳蜗毛细胞钙蛋白酶表达的影响

The Effects of Cisplatin on the Expression of Calpain in Mouse Cochlear Hair Cell in Vitro

张立伟;刘芳芳;于利;王爱梅;

1:辽宁医学院生理学教研室

摘要
目的观察顺铂对离体培养小鼠耳蜗毛细胞钙蛋白酶(calpain)表达的影响,探讨顺铂致耳蜗毛细胞凋亡的机制。方法取出生后3d的昆明小鼠300只(600耳),分离出耳蜗基底膜600条,体外培养24h后,随机分为对照组和4、8、16μg/ml顺铂组,每组150条;对照组加入2ml新鲜培养基,顺铂组分别加入2ml含不同浓度顺铂(4、8、16μg/ml)的新鲜培养基,再继续培养24h后,应用Hoechst 33258荧光染色观察耳蜗毛细胞凋亡情况,并应用免疫荧光染色和免疫印迹(Western blot)技术检测calpain 1(μ-calpain)和calpain 2(m-calpain)在耳蜗毛细胞的表达。结果 4、8、16μg/ml顺铂组耳蜗毛细胞凋亡率分别为15.63%±0.20%、38.40%±2.64%和64.24%±0.05%,均高于对照组(5.55%±0.12%),呈现明显的量效关系;不同浓度顺铂组μ-calpain和m-calpain的表达均较对照组明显增强(P<0.01),且m-calpain的表达随顺铂浓度的增高而明显增强(P<0.01)。结论顺铂可通过calpain通路诱导小鼠耳蜗毛细胞凋亡,从而发挥毒性效应。
关键词
顺铂;耳蜗;毛细胞;钙蛋白酶;小鼠
基金项目(Foundation):
辽宁省自然科学基金(2014022029)资助
作者
张立伟;刘芳芳;于利;王爱梅;
参考文献

1 Neuhof C,Neuhof H.Calpain system and its involvement in myocardial ischemia and reperfusion injury[J].World J Cardiol,2014,6:638.

2 Chang L,Wang AM.Calpain mediated cisplatin-induced ototoxicity in mice[J].Neural Regen Res,2013,8:1995.

3 Branquinha MH,Marinho FA,Sangenito LS,et al.Calpains:potential targets for alternative chemotherapeutic intervention against human pathogenic trypanosomatids[J].Curr Med Chem,2013,20:3174.

4 Yu L,Tang H,Jiang XH,et al.Involvement of calpain-I and microRNA34in kanamycin-induced apoptosis of inner ear cells[J].Cell Biol Int,2010,34:1219.

5 Momiyama J,Hashimoto T,Matsubara A,et al.Leupeptin,a calpain inhibitor,protects inner hair cells from aminoglycoside ototoxicity[J].Tohoku Journal of Experimental Medicine,2006,209:89.

6 Coffin AB,Williamson KL,Mamiya A,et al.Profiling drug-induced cell death pathways in the zebrafish lateral line[J].Apoptosis,2013,18:393.

7 Ladrech S,Guition M,Saido T,et al.Calpain activity in the amikacin-damaged rat cochlea[J].J Comp Neurol,2004,477:149.

8 Jiang H,Sha SH,Forge A,et al.Caspase-independent pathways of hair cell death induced by kanamycin in vivo[J].Cell Death and Differention,2006,13:20.

9 王苹,杜波,杜宝东.钙蛋白酶活性及其诱导内耳细胞死亡[J].国际耳鼻咽喉头颈外科杂志,2006,30:354.

本文信息

PDF(181K)

本文关键词相关文章

顺铂耳蜗毛细胞钙蛋白酶小鼠

本文作者相关文章

张立伟刘芳芳于利王爱梅