安神定志灵对大鼠脑突触体多巴胺释放相关因子表达的影响  

Effect of Anshen Dingzhi Ling on Expression of Dopamine-related Factors in Rat Brain Synaptosomes

作　　者：周荣易 孙继超[1] 尤月[1] 袁海霞[1] 宋宇尘 王娇娇[1] 韩新民[1] 

机构地区：[1]南京中医药大学中医儿科研究所,江苏南京210023

出　　处：《中药新药与临床药理》2017年第6期708-713,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金面上项目(81273801);江苏省教育厅研究生创新工程项目(KYZZ16_0410)

摘　　要：目的研究安神定志灵对突触相关蛋白25(SNAP25)、突触融合蛋白1a(Synataxin 1a)的调控作用,从多巴胺释放途径探讨安神定志灵治疗注意力缺陷多动障碍(ADHD)的作用机制。方法自发性高血压大鼠(Spontaneously hypertensive rats,SHR)随机分为模型组,盐酸哌甲酯组(10.5μg·kg^(-1)),安神定志灵低、中、高剂量(6.75,13.35,26.70 g·kg^(-1))组,每组10只,另取10只正常血压大鼠(Wistar-Kyoto rats,WKY)作为正常组,连续灌胃给药4周。采用Percoll密度梯度离心法制备脑突触体;采用Western Blot法检测突触体SNAP25、Synataxin 1a蛋白表达;Real-time PCR法检测SNAP25、Synataxin 1a m RNA表达;免疫组化法检测纹状体SNAP25、Synataxin 1a阳性细胞表达。结果与正常组比较,模型组大鼠脑突触体SNAP25及Synataxin 1a的蛋白、m RNA、阳性细胞表达量均显著降低,差异有统计学意义(P<0.05,P<0.01)。与模型组比较,各给药组的SNAP25、Synataxin 1a蛋白相对表达量均显著升高(P<0.05,P<0.01);安神定志灵低、中剂量组的Synataxin1a m RNA相对表达量显著升高(P<0.05,P<0.01),安神定志灵中、高剂量组的SNAP25 m RNA相对表达量亦显著上调(P<0.01);安神定志灵低、中剂量组的Synataxin 1a阳性细胞表达量显著升高(P<0.01),安神定志灵中、高剂量组的SNAP25阳性细胞表达量亦显著升高(P<0.01)。结论安神定志灵能够显著改善SHR大鼠脑内SNAP25、Synataxin 1a的表达,其治疗ADHD的药效机制可能与调控多巴胺释放因子相关。Objective To investigate the role of Anshen Dingzhi Ling in the regulation of synaptic-associated protein25（SNAP25）and synaptic protein 1 a（Synataxin 1 a）;and to explore the mechanism of Anshen Dingzhi Ling treatment on attention deficit hyperactivity disorder（ADHD） from dopamine release pathway. Methods The rats were randomly divided into model group,methylphenidate hydrochloride group（10.5 μg·kg-1）,Anshen Dingzhi Ling low,medium and high dose groups（6.75,13.35,26.70 g·kg-1）,10 rats in each group;along with 10 WKY rats were taken as normal group. All rats were given gavage with correspondent drugs for 4 weeks. Brain synaptosome was prepared with Percoll density gradient centrifugation. The expression of SNAP25 and Synataxin 1 a protein was detected by Western Blot method. The expression of SNAP25 and Synataxin 1 a m RNA was detected by Real-time PCR. Expression of SNAP25, Synataxin 1 a in striatal positive cells was detected by immunohistochemistry. Results Compared with the normal group, the expression levels of protein, m RNA of SNAP25 and Synataxin 1 a, and positive cells in the rat brain synaptosomes were significantly lower in the model group（P〈0.05,P〈0.01）. Compared with the model group,the expression levels of SNAP25 and Synataxin 1 a protein were significantly increased in each administration group（P〈0.05,P〈0.01）. The expression levels of Synataxin 1 a m RNA in Anshen Dingzhi Ling low and middle dose groups were significantly increased（P〈0.01）. The expression of SNAP25 m RNA was also significantly up-regulated in the Anshen Dingzhi Ling medium and high dose groups（P〈0.01）. The expression of Synataxin 1 a positive cells in the Anshen Dingzhi Ling low and middle dose groups were significantly higher（P〈0.01）. The expression of SNAP25 positive cells in the Anshen Dingzhi Ling medium and high dose groups was also significantly increased（P〈0.01）.Conclusion Anshen Dingzhi Ling can significantly improve the expression of SNAP25 and Synataxin 1 a in t

关 键 词：安神定志灵 注意力缺陷多动障碍 突触相关蛋白25 突触融合蛋白1a 多巴胺 

分 类 号：R285.5[医药卫生—中药学]