川芎嗪对豚鼠耳蜗毛细胞内酸性磷酸酶的影响

Effects of ligustrazine on acid phosphatase in the cochlear hair cell of the guinea pigs

目的:探讨活血祛淤中药制剂成分川芎嗪对庆大霉素耳中毒豚鼠耳蜗毛细胞酸性磷酸酶的保护作用。方法:实验于2004-12/2005-01在泰山医学院听觉研究室进行。选用耳耳郭反射正常的健康杂色豚鼠30只4月龄,体质量为250～300g,雌雄不拘。随机分成3组,每组10只。对照组给予肌肉注射生理盐水2mL/(kg·d);庆大霉素组给予肌肉注射硫酸庆大霉素80mg/(kg·d);川芎嗪组给予腹腔注射川芎嗪6mg/(kg·d),同时肌肉注射硫酸庆大霉素80mg/(kg·d)。各组连续用药20d。用电位仪检测用药前及用药后脑干听觉诱发电位反应阈值(与用药前比较,其值升高可提示听觉受损);用组织化学方法检测耳蜗毛细胞内酸性磷酸酶的变化(酸性磷酸酶染色缺失越明显,提示毛细胞损害程度越重)。结果:所有动物均进入结果分析。①脑干听觉诱发电位反应阈值结果:用药前对照组、庆大霉素组、川芎嗪组脑干听觉诱发电位反应阈值差异无显著性意义([36.12±0.25,35.72±2.08,35.22±1.07)dBpeSLP,P>0.05];用药20d三组脑干听觉诱发电位反应阈值分别为(37.27±0.13,69.06±24.73,40.46±10.34)dBpeSLP,即庆大霉素组与对照组和川芎嗪组相比较脑干听觉诱发电位反应阈值明显偏高,(t=5.75,t=4.77,P<0.01),川芎嗪组与对照组比较无显著性差异(t=1.38,P>0.05)。②耳蜗毛细胞内酸性磷酸酶的染色变化结果:光镜下观察耳蜗铺片显示对照组豚鼠耳蜗毛细胞排列整齐,酸性磷酸酶染色均匀,呈棕褐色;庆大霉素致聋豚鼠耳蜗毛细胞酸性磷酸酶染色变化依动物耳聋程度不同而呈现不同的表现,出现显色变淡、消失;川芎嗪组酸性磷酸酶染色变化较轻。结论:川芎嗪能明显降低庆大霉素对听觉系统的损伤,其机制可能与川芎嗪能维持溶酶体的完整性、防止毛细胞溶酶体内的水解酶逸出造成的毛细胞自溶性损伤有关。

AIM： To explore the protective effects of ligustrazine that promoting blood circulation by removing blood stasis on acid phosphatase in the cochlear hair cell of the guinea pigs with ototoxicity induced by gentamicin. METHODS： The experiment was conducted in the hearing research room of Taishan Medical University from December 2004 to January 2005. Healthy mottle guinea pigs with normal auricle reflex （250 g to 300 g in body mass, male or female） aged 4 months old were randomly divided into three groups with ten in each group： The guinea pigs in control group were intramuscularly injected with physiological saline in a dosage of 2 ml/kg per day; The guinea pigs in the gentamicin group were treated with gentamicin in a dosage of 80 mg/kg per day; Those in the ligustrazine group were intramuscularly injected with ligustrazine at a dosage of 6 mg/kg per day by intraperitoneal injection, meanwhile, 80 mg/kg ampulen per day was injected intramuscularly. The drugs were continuously given for 20 days in every group. The reactive liminal value of auditory evoked potential was detected with potential meter before and after medication （compared with that before medication, the increase of its value could indicated the hearing damage）; The changes of acid phosphatase in the cochlear hair cell with histochemical method （the more obvious the staining deletion of acid phosphatase, indicated the more severe the damage degree of hair cells）. RESULTS： All the animals were involved in the result analysis. ① Reactive liminal value of auditory evoked potential： There was no obvious significance in the control group, gentamicin group and ligustrazine group before medication [ （ 36.12±0,25,35.72±2.08, 35.22±1.07） dB peSLP, P 〉 0.05]. The reactive liminal value of auditory evoked potential in the three groups after 20 days midication was （37.27±0.13,69.06±24.73,40.46±10.34） dBpeSLP, respectively. The value in the gentamicin group was significantly higher compared with that in the control group and ligustrazine group （t=5.75, t=4.77, P 〈 0.01 ）, There was insignificant difference between the ligustrazine group and control group （t = 1.38, P 〉 0.05 ）.② Staining changes of acid phosphatase in the cochlear hair cell： To observe the cochlear stretched preparation under light microscope, which indicated that the cochlear hair cells lined up in order in the control group, the staining of acid phosphatase was even with brown; The staining changes of acid phosphatase in cochlear hair cells of guinea pigs with deafness induced by gentamicin showed different manidestation according to the different degree of deafness, appearing light color and became disappeared. The staining changes of acid phosphatase in ligustrazine were slight. CONCLUSION： The ligustrazine can reduce significantly the damage of gentarnicin on auditory system. Its mechanism may have relation with themaintenance of lysosomal integrity and prevention of the autolytic damage of hair cells induced by the overflow of hydrolase in lysosome of hair cell.