补气剂对豚鼠庆大霉素耳、肾功能损害的干预作用

Interventional Effects of Invigorating Qi Agent on Gentamycin-induced Guinea Pigs’ Ototoxic and Nephrotoxic Damages

目的探讨补气剂对豚鼠庆大霉素(GM)耳、肾功能损害的作用及其机制。方法选择健康杂色成年豚鼠36只,随机分为正常组、模型组、补气组各12只,均饲养11天。正常组常规饲养;模型组注射GM[200 mg/(kg·d)]至第9天,每天2次;补气组预先使用补气剂[1.428g/(kg·d)]10天,人工灌服及自动饮用各半,同时注射GM[200 mg/(kg·d)]至第9天,补气剂共使用21天。每组随机选取6只豚鼠于第5、7、9天检测听性脑干反应(ABR);另外6只豚鼠于11天后检测耳蜗毛细胞形态学。11天后检测血清BUN、Cr;肾脏切片苏木素-伊红染色了解肾组织病理变化;Western Blot检测肾组织ASK-1、Trx、HSF-1、Caspase-3蛋白。结果全耳蜗铺片结果显示模型组各回三排外毛细胞损毁严重,肾组织病理显示肾小球弥漫性增大,近曲小管上皮细胞明显浊肿,补气组有所改善。与正常组同期比较,模型组第5、7、9天ABR阈值,血清BUN和Cr水平以及ASK-1、Caspase-3表达升高(P<0.05),各回三排外毛细胞计数以及Trx表达减少(P<0.05)。与模型组比较,补气组各回三排外毛细胞计数以及Trx、HSF-1表达增加(P<0.05),第9天ABR阈值、血清BUN和Cr含量以及ASK-1、Caspase-3表达下降(P<0.05)。与本组第5天比较,模型组和补气组第9天ABR阈值升高(P<0.05)。11天时,与本组第3回比较,模型组和补气组第1回三排外毛细胞减少(P<0.01)。结论补气剂具有一定的保护GM诱导豚鼠耳、肾毒性损害的作用,其机制与抑制氧化应激反应有关。

Objective To investigate interventional effects and possible mechanisms of Invigorating Qi Agent(IQA)against guinea pigs′gentamycin(GM)-induced auricular and renal functional damage.Methods Totally 36 healthy variegated adult guinea pigs were divided into normal group,model group,IQA group,12 in each group.The normal group was only conventionally fed until the 11th day.The model group were injected GM 200 mg/(kg·d)for 9 days,two times a day,and feed was terminated at the 11th day.The IQA group took IQA[1.428g/(kg·d),half for artificial filling and half for automatic drinking]for 10 days in advance,and then at the same time,received GM injection at the equal dose for 9 days and feed was terminated at the 11th day.6 guinea pigs in each group were randomly selected for evaluation of audiology by measuring auditory brainstem response(ABR)at 5th,7th,9th day.Another 6 guinea pigs in each group were used to detect morphology of cochlear hair cells at 11th day.Contents of blood urea nitrogen(BUN)and Creatinine(Cr)in serum were detected.Inspect renal tissue section by hematoxylin and eosin stain(HE stain)was used to investigate pathological changes of renal tissue.The expressions of thioredoxin(Trx),heat shock factor-1(HSF-1),apoptosis signal regulating kinase-1(ASK-1)and Caspase-3 proteins in renal tissue were detected by Western Blot.Results The results of stretched surface preparation of the whole cochleae showed that three rows of the outer hair cells in each turn from model group were severely damaged.The renal tissue pathology showed renal glomerulus diffuse enlargement,and proximal convoluted tubular epithelial cells displayed marked turbidity and swelling.However,IQA group showed definite improvement.By comparison with the normal group at the same time,the ABR threshold at 5th,7th,9th day,contents of BUN and Cr in serum,and expressions of ASK-1 and Caspase-3 increased in model group(P<0.05),but the numbers of three rows of the outer hair cells in each turn and expression of Trx decreased(P<0.05).By comparison with the model group,the numbers of three rows of the outer hair cells in each turn and expression of Trx and HSF-1 increased in IQA group(P<0.05),the ABR threshold at 9th day,contents of BUN and Cr in serum,and expressions of ASK-1 and Caspase-3 decreased in IQA group(P<0.05).By comparison with 5th day at the same group,the ABR threshold at 9th day increased in model group or IQA group(P<0.05).By comparison with the third turn of the same group,the numbers of first rows of the outer hair cells in third turn decreased in model group and IQA group at 11th day(P<0.05).Conclusions IQA had definitely protective effect on GM-induced ototoxic and nephrotoxic damages.Its mechanism was related to inhibiting oxidative stress reaction.