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  本文選題：C57BL/6J小鼠　切入點：氨基糖甙類抗生素　出處：《華中科技大學》2011年碩士論文

【摘要】：目的研究卡那霉素和速尿誘導(dǎo)的耳毒性發(fā)生后CaV1.3鈣離子通道蛋白在內(nèi)耳含量的改變,以及三種細胞凋亡因子cyt-c、caspase-3、p53在內(nèi)耳中的表達改變,并分析其與卡那霉素聯(lián)合速尿快速誘導(dǎo)耳蝸損傷導(dǎo)致細胞凋亡之間的關(guān)系。 方法選取4周齡C75BL/6J小鼠為實驗對象,將小鼠隨機分為實驗組和對照組,實驗組采用硫酸卡那霉素頸部皮下注射(1g/kg),30分鐘后速尿腹腔注射(0.4g/kg);對照組給與等量生理鹽水。在注射藥物前以ABR檢查小鼠聽覺功能,注射1周后再檢測ABR反應(yīng)閾值。應(yīng)用RT-PCR(逆轉(zhuǎn)錄聚合酶鏈反應(yīng))檢測CaV1.3 mRNA在實驗組和對照組中的表達水平。采用免疫組織化學方法確定cyt-c、caspase-3、p53蛋白在耳蝸內(nèi)的定位,并比較其在實驗組和對照組之間的差別。 結(jié)果給藥后實驗組ABR的反應(yīng)閾值較對照組明顯上升,兩組之間的差異具有統(tǒng)計學意義(P0.001)。實驗組CaV1.3 mRNA的表達量較對照組有所減少。免疫組織化學檢測發(fā)現(xiàn)CaV1.3鈣通道蛋白主要分布在血管紋、螺旋韌帶、螺旋凸、毛細胞等處,在耳毒性發(fā)生后,其表達明顯減弱。cyt-c、caspase-3、p53主要分布在血管紋、螺旋韌帶、螺旋凸、毛細胞等部位,在實驗組小鼠的耳蝸其表達較對照組的表達顯著增強。 結(jié)論應(yīng)用卡那霉素聯(lián)合速尿序貫注射能引起小鼠的急性耳蝸損傷和聽力的減退。這種藥物引起的耳毒性與內(nèi)耳CaV1.3鈣離子通道蛋白的損害呈現(xiàn)相關(guān)性,并且與內(nèi)耳毛細胞、血管紋、螺旋神經(jīng)元的凋亡有關(guān)。
[Abstract]:Objective to study the changes of CaV1.3 calcium channel protein in the inner ear after ototoxicity induced by kanamycin and furosemide, and the expression of three apoptosis factors, cyt-caspase-3 and p53, in the inner ear.The relationship between apoptosis induced by kanamycin combined with furosemide and cochlear injury was analyzed.Methods 4-week-old C75BL/6J mice were randomly divided into experimental group and control group. The experimental group was injected with kanamycin sulfate subcutaneously for 30 minutes and the control group was given the same amount of normal saline.The auditory function of mice was examined by ABR before injection, and the threshold of ABR reaction was measured 1 week after injection.RT-PCR was used to detect the expression of CaV1.3 mRNA in experimental group and control group.The localization of cyt-caspase-3 p53protein in cochlea was determined by immunohistochemical method, and the difference between experimental group and control group was compared.Results the response threshold of ABR in the experimental group was significantly higher than that in the control group, and the difference between the two groups was statistically significant (P 0.001).The expression of CaV1.3 mRNA in the experimental group was lower than that in the control group.Immunohistochemical examination showed that CaV1.3 calcium channel protein was mainly distributed in stria vascularis, helical ligament, helical convex, hair cell, etc. After ototoxicity, the expression of Caspase-3 p53 was significantly decreased in stria vascularis, helical ligament, helical convexity, etc.The expression of hair cells in the cochlea of the experimental group was significantly higher than that in the control group.Conclusion kanamycin combined with sequential furosemide injection can induce acute cochlear damage and hearing loss in mice.The ototoxicity induced by the drug is related to the damage of CaV1.3 calcium channel protein in the inner ear and apoptosis of hair cells vascular striae and spiral neurons in the inner ear.
【學位授予單位】：華中科技大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R764

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