EGCG对阿米卡星诱导的大鼠螺旋神经元凋亡的保护作用

The Protection of EGCG from Amikacin-induced Apoptosis of Spiral Ganglion Neurons in Rats

罗欣荣;谢鼎华;刘强和;

1:中南大学耳科研究所

2:桂林医学院附属医院耳鼻咽喉科

摘要
目的研究硫酸阿米卡星(Amikacin sulfate)诱导SD大鼠螺旋神经元(spiral ganglion neurons,SGNs)损伤的机制及表没食子儿茶素表没食子酸酯[(-)-epigallocatechin-3-gallate,EGCG]对硫酸阿米卡星耳毒性的保护作用。方法60只SD大鼠(雌雄各半)随机均分为三组,对照组(NS组)每日皮下注射生理盐水4.5ml/kg,中毒组(A组)每日皮下注射Amikacin 450 mg/kg,保护组(AE组)每日皮下注射Amikacin 450 mg/kg,同时腹腔注射EGCG 50 mg/kg,以上三组均连续用药14天。每组用药前和用药后第7、14、21、35天随机抽取4只行ABR检测和组织学观察,比较三组间ABR的波潜伏期(peak latency,PL)和波间期(interpeak latency,IPL)、HE染色切片中的螺旋神经元(spiral ganglion neurons,SGNs)计数、TUNEL染色切片后的SGNs凋亡阳性细胞数。结果①用药前三组间ABR的PL和IPL无显著性差异,中毒组使用阿米卡星后第14和21天ABR的波潜伏期和波间期较保护组及对照组明显延长(P<0.05);②使用阿米卡星后,SGNs细胞数随时间的延长逐渐减少,保护组和中毒组有显著差异(P<0.05);而TUNEL检测片中,中毒组和保护组SGNs的凋亡阳性细胞数均随时间的延长而增多,中毒组明显多于EGCG保护组(P<0.05)。结论①阿米卡星能诱导大鼠耳蜗螺旋神经元的凋亡;②EGCG能减低阿米卡星诱导的大鼠耳蜗螺旋神经元的凋亡,对听功能、耳蜗螺旋神经元有很好的保护作用。
关键词
凋亡;硫酸阿米卡星;表没食子儿茶素表没食子酸酯;螺旋神经元;听性脑干反应
基金项目(Foundation):
作者
罗欣荣;谢鼎华;刘强和;
参考文献

1 Sha SH,Zajic G,Epstein CJ,et al.Overexpression of cop-per/zinc superoxide dismutase protects from Kanamycin-in-duced hearing loss[J].Audiol Neurootol,2001,6:117.

2刘国辉,谢鼎华,伍伟景.儿茶素对SD大鼠卡那霉素耳神经毒性保护作用的形态学研究[J].湖南医科大学学报,2002,27:503.

3 Xie D,Liu G,Zhu G,et al.(-)-Epigallocatechin-3-gal-late protects cultured spiral ganglion cells from H2O2-inducedoxidizing damage[J].Acta Otolaryngol,2004,124:464.

4 Alam SA,Ikeda K,Oshima T,et al.Cisplation-induced ap-optotic cell death in Mongolian gerbil cochlea[J].Hear Res,2000,141:28.

5 Gavrieli Y,Sherman Y,Ben-Sasson SA.Identification ofprogrammed cell death in situ via specific labeling of nuclearDNA fragmentation[J].J cell Biol,1992,119:493.

6 Lang H,Liu C.Apoptosis and hair cell degeneration in thevestibular sensory epithelia of the guinea pig following a genta-micin insult[J].Hear Res,1997,111:177.

7 Charriaut-Marlangue C,Ben-Ari Y.A Cautionary note onthe use of the TUNEL stain to determine apoptosis[J].Neu-roreport,1995,7:61.

8 Bicknell GR,Cohen GM.Cleavage of DNA to large Kilobasepair fragments occurs in some forms of necrosis as well as ap-optosis[J].Biochem Biophys Res Commun,1995,207:40.

9 Srinivas P,Gopinath G,Banerji A,et al.Plumbagin inducesreactive oxygen species,which mediate apoptosis in human cer-vical cancer cells[J].Mol Carcinog,2004,40:201.

10 Takahashi A,Masuda A,Sun M,et al.Oxidative stress-induced apoptosis is associated with alterations in mitochon-drial caspase activity and Bcl-2-dependent alteration in mi-tochondrial Ph(PHM)[J].Brain Res Bull,2004,15:497.

11 Buccellato LJ,Tso M,Akinci OI,et al.Reactive oxygenspecies are required for hyperoxia-induced Bax activationand cell death in alveolar epithelial cells[J].J Biol Chem,2004,20:6 753.

12 Simbula G,Pibiri M,Sanna L,et al.The Peroxisome prolif-erator BR931 Kills Fao cells by P53-dependent apoptosis[J].Life sci,2004 4:271.

13 Mc Fadden SL,Ding D,Jiang H,et al.Time course of effer-ent fiber and spiral ganglion cell degeneration following com-plete hair cell loss in chinchilla[J].Brain Res,2004,30:40.

14 Song BB,Anderson DJ,Schacht J.Protection from gentami-cin ototoxicity by iron chlelators in guinea pig in vivo[J].JPharmacol Exp Ther,1997,282:369.

15沙素华,高起学,李兴启,等.氨基甙类抗生素的耳毒性机理及其预防的新进展[J].听力学及言语疾病杂志,2001,9:180.

16 Chen L,Iee MJ,Li H,et al.Absorption,distribution,elimi-nation of tea polyphenols in rats[J].Drug Metab Dispos,1997,25:1 045.