南方医科大学学报

南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (04): 557-.

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盐酸椒苯酮胺对庆大霉素所致豚鼠听力损伤的保护作用

李波波，吴剑，陈婧，陈浩，李永贺   

  • 出版日期:2016-04-20 发布日期:2016-04-20

Protective effect of peperphentonamine injection through the otocyst against gentamicininduced cochlear damage in guinea pigs

  • Online:2016-04-20 Published:2016-04-20

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摘要： 目的观察豚鼠耳蜗自噬相关蛋白beclin1和LC3、Na+-K+-2Cl-联合转运体（NKCC1）mRNA、内皮素（ET）表达与耳蜗庆大霉 素（GM）损伤关系及盐酸椒苯酮胺（PPTA）对损伤的保护作用及其机制。方法将60只豚鼠随机分成对照组、模型组、同期治疗 组、造模后对照组、后期治疗组。对照组给NS+人工外淋巴液、模型组给GM+人工外淋巴液、治疗组给GM+PPTA（均连续给药 7 d），造模后对照组及后期治疗组均连续注射GM 7 d后，再分别行人工外淋巴液及PPTA连续注入7 d。所有豚鼠给药前均手 术行听泡置管，NS及GM（160 mg·kg-1·d-1）采用腹腔注射，人工外淋巴液及PPTA采用听泡注入。各组豚鼠完成给药后ABR测 试分析听力，检测beclin1和LC3、NKCC1 mRNA及ET-1的表达。结果模型组、造模后对照组的ABR结果差异无统计学意义 （P>0.05），但两者均明显高于其余3 组（P<0.05），同期治疗组豚鼠ABR阈值明显低于后期治疗组（P<0.05）；模型组LC3Ⅱ及 Beclin1的表达量较其余4组明显升高，后期治疗组LC3Ⅱ及Beclin1的表达量较造模后对照组降低；模型组NKCC1 mRNA表 达较其余4组明显升高（P<0.05），后期治疗组NKCC1 mRNA表达明显低于造模后对照组（P<0.05）。模型组各部位ET-1表达 较其余4组明显增高，对照组各部位ET-1表达低于其余4组，后期治疗组各部位ET-1表达较造模后对照组降低。结论PPTA可 抑制耳蜗细胞自噬、NKCC1、ET-1的表达，从而对耳蜗庆大霉素损伤起到保护作用；PPTA早期对抗庆大霉素耳蜗损伤效果更 优，提示GM可能对听觉细胞产生不可逆损伤。

Abstract: Objective To explore the relationship of gentamicin-induced cochlear damage with autophagy-related protein LC3, beclin1, Na+-K+-2Cl- cotransporter (NKCC1) mRNA and endothelin-1 (ET-1), and investigate the protective mechanism of PPTA against gentamicin-induced cochlear damage. Methods Sixty guinea pigs were randomly divided into control group (with saline and artificial perilymph injections), model group (with gentamicin and artificial perilymph injections), concurrent treatment group (with gentamicin and PPTA injections), model control group (with artificial perilymph injection 7 days after gentamicin injection) and delayed treatment group (with PPTA injection 7 days after gentamicin injection). Saline and gentamicin (160 mg/kg) were injected intraperitoneally, and artificial perilymph and PPTA were injected into the otocysts on a daily basis for 7 consecutive days. Hearing impairment of the guinea pigs was analyzed with ABR, and the protein expressions of beclin1 and LC3 in cochlear tissue were tested. The expression of NKCC1 mRNA was detected with RT-PCR, and the expression of ET-1 was detected immunohistochemically. Results The ABR thresholds in the model group and model control group were similar (P>0.05) , but significantly higher than those in the other 3 groups (P<0.05); the threshold was significantly lower in concurrent treatment group than in delayed treatment group (P<0.05). Compared with those in the other 4 groups, the expressions of LC3 II, beclin1, and NKCC1 mRNA were significantly increased in the model group (P<0.05); and those in delayed treatment group were significantly lower than those in the model control group (P<0.05). The expressions of ET-1 in the Corti organ, striavascularis and spiral ganglion were significantly higher in the model group but significantly lower in the control group than those in the other 4 groups; ET-1 expression was significantly lower in delayed treatment group than in the model control group. Conclusion PPTA offers protection against gantamicin-induced cochlear damage in guinea pigs by inhibiting cell autophagy and suppressing of NKCC1 and ET-1 expressions. Early intervention with PPTA produces better therapeutic effect, suggesting that gantamicin causes irreversible injury of the auditory cells.