川芎嗪干预庆大霉素耳毒性作用的机理研究
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摘要
目的探讨川芎嗪(tetramethylpyrazine,TMP)对庆大霉素(gentamycin,GM)耳毒性作用的防治机理。方法将36只豚鼠随机分成生理盐水组、庆大霉素造模组、川芎嗪防治组,每组各12只。从病理层面研究,利用TUNEL法标记比较3组豚鼠耳蜗螺旋神经节(spiralganglioncells,SGC)、螺旋器及侧壁(lateralwall,LW)内细胞凋亡情况。从基因水平研究,利用Real-timePCR法检测C-Jun氨基末端激酶(c-jun n-terminal kinases,JNK)的mRNA含量。从听力学角度,结合听性脑干反应(auditory brainstem response,ABR)测试观察用药前后豚鼠听力的变化。结果与川芎嗪防治组相比,庆大霉素造模组的耳蜗螺旋神经节细胞凋亡增多(P<0.05),螺旋器和侧壁细胞凋亡明显增多(P<0.001),JNK的mRNA表达明显升高(P<0.05),同时ABR阈移显著增大(P<0.01)。结论川芎嗪可明显降低庆大霉素导致的JNK转录水平上的高表达,从而有效防护庆大霉素的耳毒性,这可能是川芎嗪发挥防护作用的机制之一。
Objective To identify the protection mechanism of Tetramethy PYrazine in the intervention of Gentamic in ototoxicity. Methods Thirty-six guinea pigs were randomly divided into 3 groups, 12 in each: physiological saline group, gentamicin-made module and tetramethylpyrazine prevention group. From the pathological level, the TUNEL method was used to compare the apoptosis of cochlear spiral ganglion(SGC), spiral organ and lateral wall(LW) in the three groups of the guinea pigs. From the gene level study, the mRNA content of JNK was detected by Real-time PCR. Audiology combined with auditory brainstem response(ABR) test was used to observe the changes of hearing in the guinea pigs before and after medication. Results Compared with tetramethylpyrazine prevention group, the apoptosis of the spiral ganglion cells of the gentamicin-made group increased(P<0.05), and the apoptosis of the spiral and sidewall cells increased significantly(P<0.001). The expression of JNK mRNA was significantly increased(P<0.05), while the threshold shift of ABR was significantly increased(P<0.01) Conclusion Tetramethylpyrazine can significantly reduce the high expression of gentamicin-induced JNK transcription level, thus effectively protecting the ototoxicity of gentamicin, which may be one of the mechanisms of the protective effect of tetramethylpyrazine.
Objective To identify the protection mechanism of Tetramethy PYrazine in the intervention of Gentamic in ototoxicity. Methods Thirty-six guinea pigs were randomly divided into 3 groups, 12 in each: physiological saline group, gentamicin-made module and tetramethylpyrazine prevention group. From the pathological level, the TUNEL method was used to compare the apoptosis of cochlear spiral ganglion(SGC), spiral organ and lateral wall(LW) in the three groups of the guinea pigs. From the gene level study, the mRNA content of JNK was detected by Real-time PCR. Audiology combined with auditory brainstem response(ABR) test was used to observe the changes of hearing in the guinea pigs before and after medication. Results Compared with tetramethylpyrazine prevention group, the apoptosis of the spiral ganglion cells of the gentamicin-made group increased(P<0.05), and the apoptosis of the spiral and sidewall cells increased significantly(P<0.001). The expression of JNK mRNA was significantly increased(P<0.05), while the threshold shift of ABR was significantly increased(P<0.01) Conclusion Tetramethylpyrazine can significantly reduce the high expression of gentamicin-induced JNK transcription level, thus effectively protecting the ototoxicity of gentamicin, which may be one of the mechanisms of the protective effect of tetramethylpyrazine.
引文
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[4]郑华平,王枫,徐泉珍,等.针刺对川芎嗪防治豚鼠庆大霉素中毒性耳聋靶向作用的影响[J].中国针灸,2008,28(7):515-518.
[5]王永华,刘金洪,王枫,等.针刺及川芎嗪对庆大霉素致耳聋豚鼠螺旋神经节细胞凋亡及调控基因Bcl-2、Bax表达的影响[J].中国中西医结合耳鼻咽喉科杂志,2007,15(5):321-325.
[6]王佳,朱鹏霞,林松柳,等.川芎嗪逆转庆大霉素所致豚鼠耳肾氧化应激反应[J].医药导报,2018,37(8):939-942.
[7]高可雷,丁大连,李鹏,等.氨基糖苷类抗生素对耳蜗螺旋神经节的损害作用[J].中华耳科学杂志,2015,13(1):37-42.
[8]殷泽登,黎万荣.庆大霉素中毒性耳聋[J].国际耳鼻咽喉头颈外科杂志,2006,30(6):360-363.
[9]AuladellC.TheRoleofJNKPathwayintheProcessof Excitotoxicity Induced by Epilepsy and Neurodegeneration[M].In:Rocha L,Cavalheiro E,eds.Pharmacoresistance in Epilepsy.Springer,2013.99-113.
[10]Li L,Nevill G,Forge A.Two modes of hair cell loss from the vestibularsensoryepitheliaoftheguineapiginnerear[J].Journal of Comparative Neurology,2010,355(3):405-417.
[11]Ham YM,Yoon WJ,Park SY,et al.Quercitrin protects against oxidativestress-inducedinjuryinlungfibroblastcellsvia up-regulationofBcl-xL[J].JournalofFunctionalFoods,2012,4(1):253-262.
[12]Elmore S.Apoptpsis:a review of programmed cell death[J].Toxicol Pathol,2007,35(4):495-516.
[13]Ylikoski J,Xing-Qua L,Virkkala J,et al.Blockade of c-Jun N-terminalkinasepathwayattenuatesgentamicin-induced cochlearandvestibularhaircelldeath[J].HearRes,2002,163(1-2):71-81.
[14]WangJ,VanDeWaterTR,BonnyC,eta1.Apeptide inhibitorofc-JunN-tennlnalkinaseprotectsagainstboth aminoglycosideandacoustictrauma-inducedauditoryhair cell death and hearing loss[J]. Neurosci, 2003, 23(24):8596-8607.
[15]钟鸣.JNK/MAPK信号通路介导川芎嗪对海马神经元损伤的保护机制研究[D].广州:广州中医药大学,2012.