基于Mas/PKA/CREB通路探讨颈椎横突尖针刺法治疗后循环缺血性眩晕模型大鼠的作用机制  

Exploration of the mechanism of action of cervical transverse process tip acupuncture for the treatment of rats with posterior circulation ischemic vertigo based on the Mas/PKA/CREB pathway

作　　者：欧小燕 柳刚 杨丽芳 文林月 汪生志 李娜 杨骏 张闻东 OU Xiaoyan;LIU Gang;YANG Lifang;WEN Linyue;WANG Shengzhi;LI Na;YANG Jun;ZHANG Wendong(Graduate School of Anhui University of Chinese Medicine,Hefei 230031,China;The Second Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230061,China;The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230011,China)

机构地区：[1]安徽中医药大学研究生院,合肥230031 [2]安徽中医药大学第二附属医院 [3]安徽中医药大学第一附属医院

出　　处：《北京中医药大学学报》2024年第2期288-296,共9页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家重点基础研究发展计划(973计划)项目(No.2010CB530505);安徽省自然科学基金项目(No.1608085MH184);安徽省重大疑难疾病中西医协同攻关项目(皖中医药发展秘〔2021〕70号);全国中医药创新骨干人才培养项目(皖卫中医药发〔2019〕12号)。

摘　　要：目的基于Mas原癌基因(Mas)/蛋白激酶A(PKA)/环磷腺苷效应元件结合蛋白(CREB)通路探讨颈椎横突尖针刺法对后循环缺血性眩晕(PCIV)模型大鼠的作用机制。方法将32只雄性SD大鼠按照随机数字表法分为假手术组、模型组、尼莫地平组、电针组,每组各8只。除假手术组外,其余各组大鼠均结扎右侧颈总动脉(CCA)和锁骨下动脉(SCA)建立PCIV模型,假手术组仅剥离右侧CCA和SCA。尼莫地平组大鼠灌胃尼莫地平混悬液(12 mg/kg);电针组大鼠于第2颈椎(C2)、第4颈椎(C4)、第6颈椎(C6)横突尖针刺,于双侧C2、C6进行电针治疗,20 min/次;假手术组和模型组不予任何干预措施,连续14 d。观察各组大鼠右侧前庭神经核脑血流量、脑血流量下降率及跳台躲避时间,采用TUNEL染色法观察右侧脑干前庭神经核细胞凋亡情况,蛋白质印迹法检测右侧脑干前庭神经核Mas、蛋白激酶A⁃C亚基同工型a(PKA⁃Ca)、CREB、磷酸化CREB(p⁃CREB)、B细胞淋巴瘤(Bcl)⁃2和Bcl⁃2相关X蛋白(Bax)的蛋白表达情况。结果干预前,与假手术组对比,其余3组大鼠右侧前庭神经核脑血流量下降、右侧前庭神经核脑血流下降率升高(均P<0.05),提示造模成功。干预后,与假手术组比较,模型组大鼠右侧前庭神经核脑血流量降低、右侧前庭神经核脑血流下降率升高、右侧脑干前庭神经核细胞凋亡率及Bax蛋白表达水平升高,Bcl⁃2、Mas、PKA⁃Ca、p⁃CREB/CREB蛋白表达水平降低(均P<0.05);与模型组比较,电针组和尼莫地平组大鼠右侧前庭神经核脑血流量升高、右侧前庭神经核脑血流下降率降低、跳台躲避时间缩短、右侧脑干前庭神经核细胞凋亡率及Bax蛋白表达水平降低,Bcl⁃2、Mas、PKA⁃Ca、p⁃CREB/CREB蛋白表达水平升高(均P<0.05);与尼莫地平组比较,电针组大鼠右侧前庭神经核脑血流量升高、右侧前庭神经核脑血流下降率降低、跳台躲避时间缩短、右侧脑干Objective We aimed to explore the mechanism of action of cervical transverse process tip acupuncture in a rat model of posterior circulation ischemic vertigo(PCIV)based on the G protein⁃coupled receptor(Mas)/protein kinase A(PKA)/cyclophosphoadenosine effector binding protein(CREB)pathway.Methods According to the random number table method,32 male SD rats were divided into the sham operation group(n=8),the model group(n=8),the nimodipine group(n=8),and the electroacupuncture group(n=8).In all groups except for the sham operation group,the PCIV model was established by ligating the right common carotid artery(CCA)and the right subclavian artery(SCA).In the sham operation group,only the right CCA and the right SCA were removed and sutured directly without ligation.The nimodipine group was given intragastric administration of nimodipine suspension(12 mg/kg),and the electroacupuncture group selected C2,C4,and C6 transverse process tip for acupuncture,and performed electroacup uncture treatment on both sides of C2 and C6,20 minutes each time.There was no intervention in the sham operation group and the model group.The intervention was given once a day for 14 days.Cerebral blood flow,the decrease decline rate of cerebral blood flow,and the step⁃down avoidance time were recorded in each group.Apoptosis of neurons in the vestibular nucleus of the right brainstem was observed by TUNEL staining,and the protein expression levels of Mas,PKA⁃Ca,CREB,p⁃CREB,Bcl⁃2,and Bax in the vestibular nucleus of the right brainstem were determined by Western blotting.Results Before intervention,compared with the sham operation group,cerebral blood flow in the right vestibular nucleus was decreased and decline rate of cerebral blood flow in the right vestibular nucleus was increased in the remaining three groups(P<0.05),which suggested that the model was successfully established.After intervention,compared with the sham operation group,the model group showed lower cerebral blood flow in the right vestibular nucleus,larger decline

关 键 词：颈椎横突尖针刺 后循环缺血 眩晕 细胞凋亡 Mas原癌基因/蛋白激酶A/环磷腺苷效应元件结合蛋白通路 大鼠 

分 类 号：R255.5[医药卫生—中医内科学]