基于自噬蛋白ATG的表达探讨川芎嗪对药物性聋大鼠耳蜗自噬水平的干预机制  

Based on the expression of autophagy-related protein ATG,the intervention mechanism of ligustrazine on the level of autophagy in cochlea of drug-induced deafness rats was investigated

作　　者：冷辉[1] 刘传丽 马贤德[2] 张琦[1] 王爱平[1] LENG Hui;LIU Chuanli;MA Xiande;ZHANG Qi;WANG Aiping(Department of Otolaryngology Head and Neck Surgery,the Afiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang,Liaoning,110032,China;Liaoning University of Traditional Chinese Medicine,Shenyang,Liaoning,110032,China)

机构地区：[1]辽宁中医药大学附属医院耳鼻咽喉头颈外科,辽宁沈阳110032 [2]辽宁中医药大学,辽宁沈阳110032

出　　处：《中国耳鼻咽喉头颈外科》2023年第11期710-714,共5页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：辽宁省教育厅课题(L201901)。

摘　　要：目的采用顺铂腹腔注射的方法制作药物性聋大鼠模型,并使用川芎嗪进行干预,观察川芎嗪对耳聋模型大鼠耳蜗组织自噬相关蛋白5(ATG5)、ATG12和ATG16表达水平的影响,探讨川芎嗪对药物性聋大鼠耳蜗自噬的干预机制。方法将110只大鼠适应性饲养7 d后,随机分配成两组,空白组(n=45)和模型组(n=65),空白组和模型组分别注射等量的生理盐水和顺铂,连续7 d后,两组随机抽取5只大鼠进行模型评价。余下的空白组随机分配为空白对照组和川芎嗪对照组各20只,模型组随机分配为模型对照组,川芎嗪治疗组和抑制剂组各20只。空白对照组和模型对照组按照腹腔注射5 mg/(kg·d)的生理盐水,连续7 d。川芎嗪对照组和川芎嗪治疗组按照腹腔注射140 mg/(kg·d)的顺铂,连续7 d。抑制剂组按照腹腔注射30 mg/(kg·d)的3-MA抑制剂,连续7 d。通过光镜观察耳蜗毛细胞排列情况;免疫组化和Western-blot法检测自噬蛋白ATG5、ATG12和ATG16表达水平。结果通过听性脑干反应(ABR)模型评价检测结果显示,与空白组大鼠右耳检测数据中ABR阈值相比较,模型组ABR阈值显著提高(P<0.01)。免疫组化和Western-blot法显示,与空白对照组和川芎嗪对照组比较,模型对照组和川芎嗪治疗组大鼠耳蜗组织中自噬相关蛋白表达水平升高(P<0.05);与模型对照组比较,川芎嗪治疗组大鼠耳蜗组织中自噬相关蛋白表达水平升高(P<0.01);与模型对照组和川芎嗪治疗组比较,抑制剂组大鼠耳蜗组织中自噬相关蛋白表达水平降低(P<0.01),均有统计学意义。结论川芎嗪可能是通过上调自噬蛋白ATG5、ATG12和ATG16的表达水平,提高耳蜗组织中的自噬水平,从而实现治疗药物性聋的目的。OBJECTIVEETo replicate the rat model of drug-induced deafness by intraparitoneal injection of cisplatin,and to observe the effects of ligustrazine on the expression levels of autophag-related proteins ATG5,ATG12 and ATG16 in the cochlear tissues of rats with drug-induced deafness,and to explore the intervention mechanism of ligustrazine on autophagy in the cochlear tissues of rats with drug-induced deafness.METHODS After adaptive feeding for 7 days,110 rats were randomly divided into two groups:blank group(n-45)and model group(n=65).Blank group and model group were injected with the same amount of normal saline and cisplatin,respectively.Ater 7 consecutive days,5 rats were randomly selected from the two groups for model evaluation.The remaining blank group was randomly assigned to blank control group and ligustrazine control group with 20 mice each,and model group was randomly assigned to model control group,ligustrazine treatment group and inhibitor group with 20 mice each.Blank control group and model control group were intraperitoneally injected with 5 mg/(kg:d)normal saline for consecutive 7 days.Ligustrazine control group and ligustrazine treatment group were intraperitoneally injected with 140 mg/(kgd)cisplatin for consecutive 7 days.Inhibitor group was intraperitoneally injected with 30 mg/(kg:d)cisplatin for consecutive 7 days.The arrangement of cochlear hair cells was observed by light microscope.The expression levels of autophagy proteins ATG5,ATG12 and ATG16 were detected by immunohistochemistry and Western-blot.RESULTSSTheresultsofauditory brainstem response(ABR)model evaluation showed that compared with the ABR threshold in the right ear test data of the blank group,the ABR threshold of the model group was significantly increased(P<0.01).Immunohistochemistry and Western-blot showed that the expression level of autophagy-related proteins in cochlear tissues of rats in model control group and ligustrazine treatment group was increased compared with that in blank control group and Ligustrazine control g

关 键 词：顺铂 自噬 川芎嗪 自噬相关蛋白 

分 类 号：R285.5[医药卫生—中药学]