本文選題：順鉑 + 耳蝸��； 參考：《遼寧醫(yī)學(xué)院》2012年碩士論文

【摘要】：目的 建立成年BALB/c小鼠順鉑耳毒性模型，研究順鉑對(duì)小鼠耳蝸鈣蛋白酶（calpain）表達(dá)的影響，探討calpain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實(shí)驗(yàn)依據(jù)。 方法 65只健康成年BALB/c小鼠隨機(jī)分成對(duì)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13只，給予腹腔連續(xù)注射5d，每天1次。應(yīng)用免疫熒光染色方法、顯微圖像分析以及蛋白質(zhì)印跡技術(shù)檢測(cè)小鼠耳蝸中calpain1和calpain2的表達(dá)；同時(shí)結(jié)合聽腦干反應(yīng)（auditory brainstem response，ABR）測(cè)試，觀察用藥前后小鼠聽力的變化。 結(jié)果 1．腹腔連續(xù)注射5d后，對(duì)照組小鼠體重略有增加，而不同劑量順鉑組小鼠體重從第3d起均出現(xiàn)不同程度減少，與對(duì)照組比較均有顯著性差異（P＜0.05，P＜0.01）；并且隨著順鉑給藥劑量的增大，小鼠體重減少愈明顯（P＜0.05，P＜0.01）。 2．腹腔連續(xù)注射5d后，在各刺激頻率下，不同劑量順鉑組小鼠ABR閾移均明顯增大，與對(duì)照組比較均有顯著性差異（P＜0.01）；并且隨著順鉑給藥劑量的增大，ABR閾移亦顯著增大，呈現(xiàn)明顯的量效關(guān)系（P＜0.01）。 3．免疫熒光染色及顯微圖像分析結(jié)果顯示，對(duì)照組小鼠耳蝸外毛細(xì)胞、螺旋神經(jīng)節(jié)和血管紋中均有calpain1和calpain2的微弱表達(dá)；不同劑量順鉑組小鼠耳蝸上述各部位中calpain1和calpain2的表達(dá)均較對(duì)照組明顯增強(qiáng)（P＜0.05，P＜0.01），但順鉑組間calpain1的表達(dá)無明顯差異，而calpain2的表達(dá)則隨順鉑給藥劑量的增大而明顯增強(qiáng)（P＜0.01）。 4．蛋白質(zhì)印跡檢測(cè)及半定量分析結(jié)果顯示，對(duì)照組小鼠耳蝸calpain1和calpain2的蛋白表達(dá)水平較低；不同劑量順鉑組calpain1和calpain2的蛋白表達(dá)水平均較對(duì)照組明顯增高（P＜0.01），但順鉑組間calpain1的蛋白表達(dá)水平無顯著性差異，而calpain2的蛋白表達(dá)水平則隨順鉑給藥劑量的增大而明顯增強(qiáng)（P＜0.01）。 結(jié)論 應(yīng)用成年BALB/c小鼠能建立可靠的順鉑耳毒性模型。正常小鼠耳蝸中僅有少量calpain表達(dá)，順鉑可增強(qiáng)小鼠耳蝸中calpain1和calpain2的表達(dá)，并且隨著順鉑給藥劑量的增加，，calpain2的表達(dá)亦逐漸增強(qiáng)，提示calpain1和calpain2介導(dǎo)了順鉑的耳毒性損傷，并且calpain2在順鉑耳毒性損傷中可能起主導(dǎo)作用。
[Abstract]:Purpose The ototoxicity model of adult BALB/c mice was established to study the effect of cisplatin on the expression of calpainin in cochlea of mice, to explore the possible role of calpain in cisplatin ototoxicity, and to provide experimental evidence for clinical prevention and treatment of cisplatin ototoxic deafness. Method 65 healthy adult BALB/c mice were randomly divided into control group, cisplatin 2.5mg/kg group, cisplatin 3.5mg/kg group, cisplatin 4.5mg/kg group and cisplatin 5.5mg/kg group. The expression of calpain1 and calpain2 in cochlea of mice was detected by immunofluorescence staining, microimage analysis and Western blotting, and the hearing changes of mice before and after treatment were observed by combining with auditory brainstem response (ABR) test. Result 1. After 5 days of continuous intraperitoneal injection, the body weight of the control group increased slightly, but the weight of the mice in the different doses of cisplatin group decreased in different degrees from the 3rd day, there was significant difference compared with the control group (P < 0.05 P < 0.01), and with the increase of the dosage of cisplatin, the dosage of cisplatin increased. The weight loss of mice was more obvious (P < 0.05) and P < 0.01 (P < 0.01). 2. After 5 days of continuous intraperitoneal injection, the ABR threshold shift of mice in different doses of cisplatin increased significantly, compared with the control group (P < 0.01), and the threshold shift increased with the increase of the dosage of cisplatin. There was a significant dose-effect relationship (P < 0.01). 3. The results of immunofluorescence staining and microscopic image analysis showed that there were weak expressions of calpain1 and calpain2 in the outer hair cells of cochlea, spiral ganglion and stria vascularis in the control group. The expression of calpain1 and calpain2 in the cochlea of mice with different doses of cisplatin was significantly enhanced than that of the control group (P < 0.05 P < 0.01), but there was no significant difference in the expression of calpain1 among the cisplatin groups, while the expression of calpain2 increased significantly with the increase of the dosage of cisplatin (P < 0.01). 4. The results of Western blotting and semi-quantitative analysis showed that the expression of calpain1 and calpain2 in cochlea of control mice was low. The protein expression levels of calpain1 and calpain2 in different dose groups were significantly higher than those in the control group (P < 0.01), but there was no significant difference in the protein expression level of calpain1 between the cisplatin groups, but the protein expression level of calpain2 increased significantly with the increase of the dosage of cisplatin (P < 0.01). Conclusion A reliable ototoxicity model of cisplatin could be established in adult BALB/c mice. Only a small amount of calpain was expressed in the cochlea of normal mice. Cisplatin could enhance the expression of calpain1 and calpain2 in the cochlea of mice, and the expression of calpain2 increased with the increase of the dosage of cisplatin, suggesting that calpain1 and calpain2 mediated the ototoxicity of cisplatin. And calpain2 may play a leading role in cisplatin ototoxicity.
【學(xué)位授予單位】：遼寧醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R764

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