水杨酸钠作用后大鼠耳蜗K-Cl协同转运体家族mRNA表达研究

Expression of K-Cl cotransporter mRNA in Rat Cochlea Induced by Sodium Salicylate

黄娟;黄治物;杨琨;肖伯奎;

1:武汉大学人民医院耳鼻咽喉-头颈外科

2:武汉大学人民医院耳鼻咽喉-头颈外科

3:武汉大学人民医院耳鼻咽喉-头颈外科

4:武汉大学人民医院耳鼻咽喉-头颈外科 (武汉430060)

摘要
目的研究水杨酸钠作用后大鼠耳蜗K-Cl-协同转运体家族(K+-Cl- cotransporter,KCC)成员KCC14mRNA的表达情况,探讨KCC在水杨酸盐耳毒性中的作用及意义。方法选用30只正常大鼠,随机分成五组,每组6只:第一组为预实验组,未给予任何处理,采用逆转录PCR技术检测大鼠耳蜗中KCC14mRNA的表达情况,探讨KCC在水杨酸盐耳毒性中的作用及意义。方法选用30只正常大鼠,随机分成五组,每组6只:第一组为预实验组,未给予任何处理,采用逆转录PCR技术检测大鼠耳蜗中KCC14mRNA表达情况;第二、三、四、五组分别为对照组(不用药)、慢性组(肌肉注射水杨酸钠175mg/kg,2次/天,连用14天后处死观察)、急性组(一次肌肉注射水杨酸钠400mg/kg后处死观察)、恢复组(水杨酸钠用量及用法同慢性组,停药14天后处死观察),运用荧光定量PCR技术比较各组大鼠耳蜗内KCC14mRNA表达情况;第二、三、四、五组分别为对照组(不用药)、慢性组(肌肉注射水杨酸钠175mg/kg,2次/天,连用14天后处死观察)、急性组(一次肌肉注射水杨酸钠400mg/kg后处死观察)、恢复组(水杨酸钠用量及用法同慢性组,停药14天后处死观察),运用荧光定量PCR技术比较各组大鼠耳蜗内KCC14mRNA表达情况。结果KCC14mRNA表达情况。结果KCC14mRNA在正常成年大鼠耳蜗中均有表达;在对照组及水杨酸钠作用后大鼠耳蜗KCC14mRNA在正常成年大鼠耳蜗中均有表达;在对照组及水杨酸钠作用后大鼠耳蜗KCC14mRNA表达均有变化,急性组KCC1mRNA、慢性组和恢复组KCC2mRNA、恢复组KCC4mRNA表达水平高于对照组,差异有统计学意义(P<0.05),急性组KCC2mRNA、急性组、慢性组和恢复组KCC3mRNA以及急性组、慢性组KCC4mRNA表达水平低于对照组,差异有统计学意义(P<0.05)。结论KCC14mRNA表达均有变化,急性组KCC1mRNA、慢性组和恢复组KCC2mRNA、恢复组KCC4mRNA表达水平高于对照组,差异有统计学意义(P<0.05),急性组KCC2mRNA、急性组、慢性组和恢复组KCC3mRNA以及急性组、慢性组KCC4mRNA表达水平低于对照组,差异有统计学意义(P<0.05)。结论KCC14mRNA在正常大鼠耳蜗中均有表达,提示其可能共同参与耳蜗内淋巴液Cl-浓度的维持;在水杨酸钠作用后KCC14mRNA在正常大鼠耳蜗中均有表达,提示其可能共同参与耳蜗内淋巴液Cl-浓度的维持;在水杨酸钠作用后KCC14mRNA表达的不同变化,破坏了耳蜗内淋巴液Cl-浓度平衡,影响外毛细胞能动性。
关键词
K-Cl协同转运体;水杨酸钠;外毛细胞能动性;PCR;大鼠
基金项目(Foundation):
作者
黄娟;黄治物;杨琨;肖伯奎;
参考文献

1Chee KS,Kistler J,Donaldson PJ.Roles for KCCtransporters in the maintenance of lens transparency[J].Invest Ophthal mol Vis Sci,2006,47:673.

2Huang ZW,Luo YY,Wu ZY,et al.Paradoxical enhancement of active cochlear mechanicsinlong-termadministration of salicylate[J].J Neurophysical,2005,93:2053.

3Zheng J,Shen W,He DZ,et al.Prestin is the motor protein of cochlear outer hair cells[J].Nature,2000,405:149.

4Zheng J,Madison LD,Oliver D,et al.Prestin,the motor protein of outer hair cells[J].Audiol Neurootol,2002,7:9.

5Cazals Y,Horner KC,Huang ZW.Alterations of average spec-trumof cochleoneural activity bylong-termsalicylate treatment in guinea pig:a plausibleindex of tinnitus[J].J Neurophysical,1998,80:2113.

6Oliver D,He DZ,Kl cker N,et al.Intracellular anions as the voltage sensor of prestin,the outer hair cell motor protein[J].Sci-ence,2001,292:2340.

7Fettiplace R,Hackney CM.The sensory and motor roles of audi-tory hair cells[J].Nat Rev Neurosci,2006,7:19.

8Adragna NC,Lauf PK.K-Cl cotransport function andits poten-tial contribution to cardiovascular disease[J].Pathophysiology,2007,14:135.

9Williams JR,Sharp J W,Kumari VG,et al.The neuron-specific K-Cl cotransporter,KCC2antibody development andinitial char-acterization of the protein[J].J Biol Chem,1999,274:12656.

10黄治物,陈平,梅玲,等.急慢性水杨酸钠注射后大鼠耳蜗基因表达谱研究[J].中华耳鼻咽喉头颈外科杂志,2005,40:27.

11Boettger T,Rust MB,Maier H,et al.Loss of K-Cl co-trans-porter KCC3causes deafness,neurodegeneration and reduced sei-zure threshold[J].EMBOJ,2003,22:5422.

12Boettger T,Hbner CA,Maier H,et al.Deafness andrenal tubu-lar acidosisin mice lacking the K-Cl co-transporter KCC4[J].Nature,2002,416:874.

13Kharkovets T,Hardelin JP,Safieddine S,et al.KCNQ4,a K+channel mutatedin a formof dominant deafness,is expressed in theinner ear andthe central auditory pathway[J].Proc Natl Acad Sci USA,2000,97:4333.

14Kikuchi T,Adams JC,Miyabe Y,et al.Potassiumion recycling pathway via gap junction systems in the mammalian cochlea and itsinterruptionin hereditary nonsyndromic deafness[J].Med E-lectron Microsc,2000,33:516.

15黄治物,罗燕云,吴展元,等.水杨酸钠急慢性注射致豚鼠形态学的改变[J].武汉大学学报(医学版),2003,24:6.