CAJ | 학술논문

目的：了解铜离子转运蛋白1（copper transport protein 1，CTR1）在大鼠内耳的表达情况及硫酸铜和顺铂鼓室给药对CTR1表达的影响。方法 Wistar雄性大鼠24只，随机分为4组，每组6只，Ⅰ组为正常对照组（非处理组）；Ⅱ组为双耳圆窗龛放置顺铂溶液（0．5 mg／ml）的明胶海绵；Ⅲ组为双耳圆窗龛放置顺铂溶液（1 mg／ml）的明胶海绵；Ⅳ组双耳圆窗龛放置硫酸铜溶液（0．02 mg／kg）的明胶海绵。采用免疫组织化学技术，对各组耳蜗冰冻切片行CTR1蛋白的免疫组织化学染色，光学显微镜下观察耳蜗组织中CTR1的表达情况；提取各组耳蜗组织总蛋白，应用 Western－blot技术检测各组CTR1蛋白表达水平；提取耳蜗组织的总RNA ，应用RT －PCR技术检测各组CTR1mRNA的表达水平。结果Ⅰ～Ⅳ组大鼠的耳蜗各回Corti器、螺旋神经节、血管纹的细胞质和细胞膜上均有 CTR1表达，CTR1的平均光密度值呈降低趋势；各组耳蜗组织均有明显的 CTR1表达，Ⅰ～Ⅳ组C T R1蛋白的光密度值分别为0．532±0．031、0．394±0．024、0．234±0．030和0．191±0．015，呈下降趋势，各组间比较差异有统计学意义（P＜0．05）；各组耳蜗组织均有明显的CTR1 mRNA表达，Ⅰ～Ⅳ组光密度值分别为0．508±0．035、0．391±0．022、0．240±0．023和0．186±0．021，呈下降趋势，各组间比较差异均有统计学意义（ P＜0．05）。结论 CTR1在大鼠内耳有丰富的表达，其表达量可随鼓室内顺铂浓度增高而下降，且鼓室内给予硫酸酮其表达量下降。
목적：료해동리자전운단백1（copper transport protein 1，CTR1）재대서내이적표체정황급류산동화순박고실급약대CTR1표체적영향。방법 Wistar웅성대서24지，수궤분위4조，매조6지，Ⅰ조위정상대조조（비처리조）；Ⅱ조위쌍이원창감방치순박용액（0．5 mg／ml）적명효해면；Ⅲ조위쌍이원창감방치순박용액（1 mg／ml）적명효해면；Ⅳ조쌍이원창감방치류산동용액（0．02 mg／kg）적명효해면。채용면역조직화학기술，대각조이와빙동절편행CTR1단백적면역조직화학염색，광학현미경하관찰이와조직중CTR1적표체정황；제취각조이와조직총단백，응용 Western－blot기술검측각조CTR1단백표체수평；제취이와조직적총RNA ，응용RT －PCR기술검측각조CTR1mRNA적표체수평。결과Ⅰ～Ⅳ조대서적이와각회Corti기、라선신경절、혈관문적세포질화세포막상균유 CTR1표체，CTR1적평균광밀도치정강저추세；각조이와조직균유명현적 CTR1표체，Ⅰ～Ⅳ조C T R1단백적광밀도치분별위0．532±0．031、0．394±0．024、0．234±0．030화0．191±0．015，정하강추세，각조간비교차이유통계학의의（P＜0．05）；각조이와조직균유명현적CTR1 mRNA표체，Ⅰ～Ⅳ조광밀도치분별위0．508±0．035、0．391±0．022、0．240±0．023화0．186±0．021，정하강추세，각조간비교차이균유통계학의의（ P＜0．05）。결론 CTR1재대서내이유봉부적표체，기표체량가수고실내순박농도증고이하강，차고실내급여류산동기표체량하강。
Objective To study the expression of copper transport protein 1 in the inner ear of rat and the changes of CTR1 expression after those round window niche copper sulfate and cisplatin infusion .Methods 24 male wistar rats were randomly divided into 4 groups :Group I as the normal control group (nontreatment group);Group II as the round window niche cisplatin infusion group(0 .5 mg/ml);Group III as the round window niche cisplatin infusion group (1 mg/ml);group IV as the round window niche copper sulfate infusion (0 .02 mg/kg) .The CTR1 protein was detected by the immunohistochemical (IHC) otaining and Western-blot ,and the CTR1mRNA expres‐sion levels were detected by RT -PCR .Results The expression of CTR1 protein was observed in the cytoplasm and cell membrane of Corti organ cells ,spiral ganglion cells and stria vascularis in all groups .The average optical densi‐ties of CTR1 protein was a downward trend .The expression of CTR1 protein was observed in four different groups . The optical density analysis of CTR1 showed that the optical densities were 0 .532 ± 0 .031 ,0 .394 ± 0 .024 ,0 .234 ± 0 .030 and 0 .191 ± 0 .015 ,respectively .There was a downward trend ,and there were statistically differences among the groups (P<0 .05) .The CTR1 mRNA was observed in all groups .The optical density analysis of CTR1 mRNAshowed that the optical densities were 0 .508 ± 0 .035 ,0 .391 ± 0 .022 ,0 .240 ± 0 .02 and 0 .186 ± 0 .021 ,respective‐ly .It had a downward trend and were statistically differences among the groups (P<0 .05) .Conclusion The CTR1 protein was abundantly expressed in Corti organ ,spiral ganglion cells and stria vascularis of the cochlea .The round window cisplatin and copper sulfate infusion can change the expression of CTR1 proteins in inner ear .