通窍益智颗粒通过cAMP/PKA-CREB信号通路影响血管性痴呆大鼠海马神经元凋亡的研究  

Effect of Tongqiaoyizhi Granule on Apoptosis of Hippocampal Neurons in Rats with Vascular Dementia through cAMP/PKA-CREB Signal Pathway

作　　者：李双阳 王凌雪 蒲玉婷 王饶琼 唐红梅 白雪 Li Shuangyang;Wang Linxue;Pu Yuting;Wang Raoqiong;Tang Hongmei;Bai Xue(Department of Neurology,The Affiliated Traditional Medicine Hospital of Southwest Medical University,Sichuan 646000;The Affiliated Traditional Medicine Hospital of Southwest Medical University,Sichuan 646000)

机构地区：[1]西南医科大学附属中医医院神经内科,泸州646000 [2]西南医科大学附属中医医院,泸州646000

出　　处：《中药药理与临床》2020年第2期190-195,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家中医药管理局中医药防治重大疑难疾病能力项目[川财社(2014)167号];四川省教育厅项目[川教函(2016)683]。

摘　　要：目的:研究通窍益智颗粒对血管性痴呆模型大鼠的作用机制。方法:将60只大鼠随机选取10只作为假手术对照组,其余大鼠以双侧颈总动脉永久结扎法制备血管性痴呆大鼠模型,随机分为模型对照组、尼莫地平6.25 mg/kg阳性对照组和通窍益智颗粒24.8、12.4、6.2 g/kg剂量组,灌胃30 d后,观察大鼠行为学改变、TUNEL法检测海马CA1神经元凋亡率,免疫组织化学法检测即刻早期基因(c-fos)表达,Elisa法检测神经元环磷酸腺苷(cAMP)含量,Western Blot检测激酶A(PKA)、环磷酸腺效应元件结合蛋白(CREB)和磷酸化环磷酸腺苷效应元件结合蛋白(pCREB)的表达。结果:与假手术对照组比较,模型对照组潜伏逃避期明显延长,且穿过原平台次数及游泳时间明显缩短,海马CA1神经元凋亡率和c-fos表达量明显提高,cAMP、PKA和pCREB的蛋白表达水平明显下调(P<0.05);尼莫地平组和通窍益智颗粒24.8、12.4、6.2 g/kg组大鼠学习记忆能力较模型对照组明显提高,海马CA1神经元凋亡减少,c-fos的表达降低,cAMP含量、PKA及pCREB蛋白水平上调(P<0.05),其中以通窍益智颗粒24.8 g/kg效果最佳(P<0.05)。结论:基于开通玄府法在组方中重用风药组成的通窍益智颗粒,可显著改善血管性痴呆大鼠的学习记忆能力,其机制可能与调控cAMP/PKA信号通路关键因子的表达,减轻神经元凋亡有关。Objective:To study the mechanism of Tongqiaoyizhi granule on the prevention and treatment for vascular dementia rat model.Methods:10 rats were randomly selected from 60 rats as the sham operation group,other 50 rats were used to establish the rat model with vascular dementia by permanent bilateral common carotid artery ligation method.After modeling,rats were randomly divided into the model group,nimodipine positive control group(6.25 mg/kg)and Tongqiaoyizhi granule groups(24.8,12.4,6.2 g/kg).After 30 days of administration,the behavioral changes of rats were observed and the apoptosis rate of hippocampal CA1 neurons was detected by TUNEL method.The expression of c-fos was detected by immunohistochemistry,and the content of cAMP in neurons was detected by ELISA.The expressions of PKA,CREB and pCREB were detected by Western Blot.Results:Compared with the sham group,the escape latency in the model group was significantly prolonged,and the times of crossing the virtual platform and swimming duration were shortened.The apoptosis rate and c-fos expression of hippocampal CA1 neurons were increased,while the levels of cAMP,PKA and pCREB were significantly decreased(P<0.05).Compared with the model group,the learning and memory abilities of rats in the positive control group and all doses of Tongqiaoyizhi granule groups were significantly improved,the apoptosis rate of hippocampal CA1 neurons was decreased,the expression of c-fos was decreased,and the cAMP content,the levels of PKA and pCREB protein were up-regulated in varying degrees(P<0.05).Among all drugs,the effect of 24.8 g/kg Tongqiaoyizhi granule was the most significant(P<0.05).Conclusion:Based on"Kaitongxuanfu"TCM theory,"Feng"drugs played important roles in Tongqiaoyizhi granule.Tongqiaoyizhi granule can significantly improve the learning and memory ability of rats with vascular dementia,and its mechanism may be related to the regulation of the expression of key factors in cAMP/PKA signal pathway,thus reducing the neuronal apoptosis.

关 键 词：通窍益智颗粒 血管性痴呆 环磷酸腺苷 环磷腺苷效应元件结合蛋白 凋亡 即刻早期基因 

分 类 号：R285.5[医药卫生—中药学]