顺铂对小鼠耳蜗TRPA1和TRPV1表达的影响

发布时间：2018-03-01 02:02

本文关键词： 顺铂耳蜗瞬时受体电位通道蛋白A1 瞬时受体电位通道蛋白V1 免疫荧光　出处：《辽宁医学院》2012年硕士论文　论文类型：学位论文

【摘要】：目的研究顺铂对小鼠耳蜗瞬时受体电位通道蛋白A1（transient receptor potentialA1，TRPA1）和瞬时受体电位通道蛋白V1（transient receptor potential vanilloid1，TRPV1）表达的影响，探讨TRPA1和TRPV1在顺铂耳毒性中的可能作用，为临床防治顺铂耳毒性聋提供新的方法和理论依据。方法 70只健康成年BALB/c小鼠随机分成对照组、顺铂2.5mg/kg组、顺铂3.5mg/kg组、顺铂4.5mg/kg组和顺铂5.5mg/kg组，每组14只，腹腔连续注射5d，每天1次。应用免疫荧光染色方法、显微图像分析和蛋白质印迹技术检测TRPA1和TRPV1在小鼠耳蜗中的表达；同时结合听脑干反应(auditorybrainstem response，ABR)测试，测定用药前后小鼠听力的变化。结果 1．腹腔连续注射5d后，在各刺激频率下，不同浓度顺铂组小鼠ABR阈移均明显增大，与对照组比较均差异显著（P＜0.01）；并且随着顺铂浓度的增加，ABR阈移亦显著增大，呈现明显的量效关系（P＜0.01）。 2．免疫荧光染色及显微图像分析结果显示，对照组小鼠耳蜗外毛细胞、螺旋神经节以及血管纹中均有TRPA1和TRPV1的微弱表达；不同浓度顺铂组小鼠耳蜗上述各部位中TRPA1和TRPV1的表达均较对照组明显增强（P＜0.05，P＜0.01），但顺铂组间TRPA1的表达无明显差异，而TRPV1的表达则随顺铂浓度的增大而明显增强（P＜0.05，P＜0.01）。 3.蛋白质印迹检测及半定量分析结果显示，对照组小鼠耳蜗中TRPA1与TRPV1的蛋白表达水平较低；不同浓度顺铂组TRPA1和TRPV1的蛋白表达水平均较对照组明显增高（P＜0.05，P＜0.01），但顺铂组间TRPA1的蛋白表达水平无明显差别，而TRPV1的蛋白表达水平则随顺铂浓度的增大而明显增强（P＜0.05，P＜0.01）。结论正常BALB/c小鼠耳蜗中仅有TRPA1和TRPV1的少量表达。顺铂可增强TRPA1和TRPV1在小鼠耳蜗中的表达，并且随着顺铂浓度的增大，，TRPV1的表达亦逐渐增强，提示TRPA1和TRPV1介导了顺铂的耳毒性损伤，并且TRPV1在顺铂耳毒性损伤中可能起主导作用。
[Abstract]:Purpose. To investigate the effects of cisplatin on the expression of transient receptor potentialA1 (TRPA1) and transient receptor potential vanilloid1 (TRPV1) in mouse cochlea, and to explore the possible role of TRPA1 and TRPV1 in cisplatin ototoxicity. To provide a new method and theoretical basis for clinical prevention and treatment of cisplatin ototoxic deafness. Method. 70 healthy adult BALB/c mice were randomly divided into control group, cisplatin 2.5 mg / kg group, cisplatin 3.5 mg / kg group, cisplatin 4.5 mg / kg group and cisplatin 5.5 mg / kg group. The expression of TRPA1 and TRPV1 in mouse cochlea was detected by microimage analysis and Western blotting, and the hearing changes of mice before and after treatment were determined by combining with auditory brainstem response (ABR) test. Results. 1. After 5 days of continuous intraperitoneal injection, the ABR threshold shift of mice in different concentrations of cisplatin increased significantly, compared with the control group (P < 0.01), and increased with the increase of cisplatin concentration. There was a significant dose-effect relationship (P < 0.01). 2. The results of immunofluorescence staining and microscopic image analysis showed that there were weak expressions of TRPA1 and TRPV1 in the outer hair cells of cochlea, spiral ganglion and stria vascularis in the control group. The expression of TRPA1 and TRPV1 in the cochlea of mice with different concentrations of cisplatin was significantly increased compared with the control group (P < 0.05 P < 0.01), but there was no significant difference in the expression of TRPA1 among the cisplatin groups, while the expression of TRPV1 increased significantly with the increase of the concentration of cisplatin (P < 0.05 P < 0.01). 3.The results of Western blotting and semi-quantitative analysis showed that the protein expression of TRPA1 and TRPV1 in cochlea of control mice was lower than that of control mice. The protein expression levels of TRPA1 and TRPV1 in cisplatin group were significantly higher than those in control group (P < 0.05 P < 0.01), but there was no significant difference between cisplatin group and cisplatin group. However, the protein expression level of TRPV1 increased significantly with the increase of cisplatin concentration (P < 0.05 P < 0.01). Conclusion. Cisplatin could enhance the expression of TRPA1 and TRPV1 in cochlea of normal BALB/c mice, and the expression of TRPV1 gradually increased with the increase of cisplatin concentration, suggesting that TRPA1 and TRPV1 mediated the ototoxicity of cisplatin. And TRPV1 may play a leading role in cisplatin ototoxicity.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R764.43

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