阿米替林通过Ca^(2+)/CaMKⅡ/CREB信号通路改善模型大鼠耳鸣症状的机制  

The Mechanism of Amitriptyline in Improving Tinnitus Symptoms Through Ca^(2+)/CaMKⅡ/CREB Signaling Pathway in Model Rats

作　　者：刘川 林彦涛[1] 张洪艳[1] 李栋[1] LIU Chuan;LIN Yantao;ZHANG Hongyan;LI Dong(The First Affiliated Hospital of Hebei North University,Zhangjiakou,Hebei,China 050051)

机构地区：[1]河北北方学院附属第一医院,河北张家口050051

出　　处：《中国药业》2025年第6期30-34,共5页China Pharmaceuticals

基　　金：河北省张家口市级科技计划自筹经费项目[1821053D]。

摘　　要：目的探讨阿米替林通过钙离子(Ca^(2+))/钙调素激酶Ⅱ(CaMKⅡ)/环磷腺苷效应元件结合蛋白(CREB)信号通路对模型大鼠耳鸣症状的改善作用及相关机制。方法将30只SD大鼠随机分为对照组(等体积生理盐水)、模型组(等体积生理盐水)和阿米替林组(5 mg/kg),各10只。腹腔注射水杨酸钠(400 mg/kg)以复制耳鸣大鼠模型,每天1次,连续7 d,每天建模0.5 h后腹腔注射相应药物或生理盐水。建立大鼠“背景噪声停止-舔水动作减少”的条件反射模型,记录大鼠舔水时间和舔水量;采用听觉脑干反应(ABR)测量系统检测脑干反应(ABR)阈值,荧光分光光度计检测大鼠听觉皮层细胞胞浆内Ca^(2+)水平,Western blot法检测钙调素(CaM)、N-甲基-d-天冬氨酸受体2B亚基(NR2B)、CaMKⅡ和CREB的蛋白表达水平。结果与模型组比较,阿米替林组大鼠舔水时间显著延长,舔水抑制率显著降低;ABR显著降低,大鼠听觉皮层细胞胞浆内Ca^(2+)水平显著降低,CaM及NR2B蛋白表达水平显著降低,p-CaMKⅡ/CaMKⅡ及p-CREB/CREB显著降低(P<0.05)。结论阿米替林可通过抑制模型大鼠听觉皮层细胞中的Ca^(2+)/CaMKⅡ/CREB信号通路改善耳鸣症状。Objective To explore the improvement effect of amitriptyline on tinnitus symptoms in model rats and the related mechanisms through the calcium ion(Ca^(2+))/calmodulin kinaseⅡ(CaMKⅡ)/cAMP response element-binding protein(CREB)signaling pathway.Methods Thirty SD rats were randomly divided into a control group(equal volume of normal saline),a model group(equal volume of normal saline),and an amitriptyline group(5 mg/kg),with 10 rats in each group.Sodium salicylate(400 mg/kg)was intraperitoneally injected to establish a tinnitus rat model once a day for 7 consecutive days.The corresponding drug or normal saline was intraperitoneally injected 0.5 hours after modeling each day.A conditioned reflex model of"background noise cessation-reduced licking behavior"in rats was established,and the licking time and licking volume of rats were recorded.The auditory brainstem response(ABR)measurement system was used to detect the response threshold.The fluorescence spectrophotometer was used to detect the intracellular Ca^(2+)level in the auditory cortex cells of rats.The protein expression levels of calmodulin(CaM),N-methyl-d-aspartate receptor 2B subunit(NR2B),CaMKⅡ,and CREB were detected by Western blot.Results Compared with the model group,the licking time of rats in the amitriptyline group was significantly prolonged,the licking inhibition rate was significantly decreased,the ABR threshold was significantly decreased,the Ca^(2+)level in the auditory cortex cells of rats was significantly decreased,the protein expression levels of CaM and NR2B were significantly decreased,and p-CaMKⅡ/CaMKⅡand p-CREB/CREB were significantly decreased(P<0.05).Conclusion Amitriptyline can improve tinnitus symptoms by inhibiting the Ca^(2+)/CaMKII/CREB signaling pathway in the auditory cortex cells of model rats.

关 键 词：阿米替林 钙离子/钙调素激酶Ⅱ/环磷腺苷效应元件结合蛋白信号通路 耳鸣 大鼠 

分 类 号：R965[医药卫生—药理学]