本文關鍵詞： 順鉑 耳蝸 瞬時受體電位通道蛋白A1 瞬時受體電位通道蛋白V1 免疫熒光　出處：《遼寧醫(yī)學院》2012年碩士論文　論文類型：學位論文

【摘要】：目的 研究順鉑對小鼠耳蝸瞬時受體電位通道蛋白A1（transient receptor potentialA1，TRPA1）和瞬時受體電位通道蛋白V1（transient receptor potential vanilloid1，TRPV1）表達的影響，探討TRPA1和TRPV1在順鉑耳毒性中的可能作用，為臨床防治順鉑耳毒性聾提供新的方法和理論依據(jù)。 方法 70只健康成年BALB/c小鼠隨機分成對照組、順鉑2.5mg/kg組、順鉑3.5mg/kg組、順鉑4.5mg/kg組和順鉑5.5mg/kg組，每組14只，腹腔連續(xù)注射5d，每天1次。應用免疫熒光染色方法、顯微圖像分析和蛋白質印跡技術檢測TRPA1和TRPV1在小鼠耳蝸中的表達；同時結合聽腦干反應(auditorybrainstem response，ABR)測試，測定用藥前后小鼠聽力的變化。 結果 1．腹腔連續(xù)注射5d后，在各刺激頻率下，不同濃度順鉑組小鼠ABR閾移均明顯增大，與對照組比較均差異顯著（P＜0.01）；并且隨著順鉑濃度的增加，ABR閾移亦顯著增大，呈現(xiàn)明顯的量效關系（P＜0.01）。 2．免疫熒光染色及顯微圖像分析結果顯示，對照組小鼠耳蝸外毛細胞、螺旋神經節(jié)以及血管紋中均有TRPA1和TRPV1的微弱表達；不同濃度順鉑組小鼠耳蝸上述各部位中TRPA1和TRPV1的表達均較對照組明顯增強（P＜0.05，P＜0.01），但順鉑組間TRPA1的表達無明顯差異，而TRPV1的表達則隨順鉑濃度的增大而明顯增強（P＜0.05，P＜0.01）。 3.蛋白質印跡檢測及半定量分析結果顯示，對照組小鼠耳蝸中TRPA1與TRPV1的蛋白表達水平較低；不同濃度順鉑組TRPA1和TRPV1的蛋白表達水平均較對照組明顯增高（P＜0.05，P＜0.01），但順鉑組間TRPA1的蛋白表達水平無明顯差別，而TRPV1的蛋白表達水平則隨順鉑濃度的增大而明顯增強（P＜0.05，P＜0.01）。 結論 正常BALB/c小鼠耳蝸中僅有TRPA1和TRPV1的少量表達。順鉑可增強TRPA1和TRPV1在小鼠耳蝸中的表達，并且隨著順鉑濃度的增大，，TRPV1的表達亦逐漸增強，提示TRPA1和TRPV1介導了順鉑的耳毒性損傷，并且TRPV1在順鉑耳毒性損傷中可能起主導作用。
[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.
【學位授予單位】：遼寧醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R764.43

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