补肾活血通窍汤对庆大霉素耳中毒豚鼠听性脑干反应阈值的影响及机制研究〖JZ)〗〖HS)〗

目的:探讨补肾活血通窍汤对庆大霉素耳中毒豚鼠听性脑干反应(ABR)阈值的影响及机制。方法:将50只豚鼠随机分为空白组、模型组和高/中/低剂量实验组,每组10只。模型组和实验组每日腹腔注射硫酸庆大霉素100 mg/kg构建庆大霉素耳中毒模型,空白组每日腹腔注射生理盐水25 m L/kg,同时高/中/低剂量实验组分别灌胃高/中/低剂量补肾活血通窍汤(含生药量2. 40、0. 50、0. 10 mg/m L) 0. 8 mL/只,模型组和空白组灌胃等量生理盐水,连续干预10 d。干预结束后检测各组豚鼠ABR阈值;蛋白免疫印迹法(WB)及免疫组化染色观察各组豚鼠耳蜗组织自噬相关蛋白及半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白表达情况。结果:造模后,与空白组比较,模型组豚鼠ABR阈值、耳蜗组织LC3、Beclin1蛋白表达量及螺旋神经节细胞、毛细胞及血管纹中Caspase-3蛋白表达量升高(P 0. 05);补肾活血通窍汤干预结束后,与模型组比较,中/高剂量实验组ABR阈值及耳蜗组织LC3、Beclin1蛋白表达量及螺旋神经节细胞、毛细胞及血管纹中Caspase-3蛋白表达量降低(P 0. 05),低剂量实验组上述各项指标无明显变化(P 0. 05)。结论:补肾活血通窍汤可对庆大霉素耳中毒损伤起到保护作用,其机制与抑制耳蜗细胞自噬及凋亡程序的发生有关。

Study on the Effects and Mechanism of Bushen Huoxue Tongqiao Decoction on Auditory Brainstem Response Threshold of Guinea Pigs with Gentamycin Ototoxicity

To explore the effects and mechanism of Bushen Huoxue Tongqiao Decoction on auditory brainstem response(ABR)threshold of guinea pigs with gentamycin ototoxicity.Methods:A total of 50 guinea pigs were randomly divided into a blank group,a model group and high/medium/low-dose(H/M/L-dose)experiment groups,with 10 in each group.The model group and the experimental groups were established gentamycin ototoxicity model by intraperitoneal injection of gentamycin sulfate 100 mg/kg daily,and the blank group received intraperitoneal injection of normal saline 25 mL/kg daily.Meanwhile,the H/M/L-dose experiment groups were respectively given intragastric administration of high/medium/low dose of Bushen Huoxue Tongqiao Decoction(containing raw medicine dose of 2.40,0.50 and 0.10 mg/mL)with 0.8 mL for each one.The model group and the blank group were given intragastric administration of the same amount of normal saline.The continuous intervention lasted for 10 d.After the intervention,the ABR threshold of each guinea pig was detected; and the expression of autophagy related protein and cysteine aspartic proteinase-3(Caspase-3)protein in the cochlear tissues of the guinea pigs were observed by Western blotting(WB)and immunohistochemical staining.Results:After the modeling,when compared with the blank group,the ABR threshold,the expression of LC3 and Beclin1 protein in the cochlear tissue and the expression of Caspase-3 protein in the spiral ganglion cells,hair cells and stria vascularis of the guinea pigs in the model group were increased(P<0.05).After the intervention of Bushen Huoxue Tongqiao Decoction,when compared with the model group,the ABR threshold,the expression of LC3 and Beclin1 protein in the cochlear tissue and the expression of Caspase-3 protein in the spiral ganglion cells,hair cells and stria vascularis in the M and H-dose experiment groups were decreased(P<0.05).And there was no significant difference in the above indexes of the L-dose experiment group(P>0.05).Conclusion:Bushen Huoxue Tongqiao Decoction can play a protective role in the damage of gentamicin ototoxicity,and the mechanism is related to the inhibition of autophagy and apoptosis in cochlear cells.