醒脑静注射液对大鼠全脑缺血性深昏迷的早期保护作用及其机制  

Protective effects and mechanisms of Xingnaojing Injection on early global brain ischemic-induced deep coma in rats

作　　者：信红亚[1] 石镇港 吴莉峰 张妙红 袁湘中 王平 徐永兴 曾贵荣 王海均[4] XIN Hongya;SHI Zhengang;WU Lifeng;ZHANG Miaohong;YUAN Xiangzhong;WANG Ping;XU Yongxing;ZENG Guirong;WANG Haijun(Department of Blood,Xiangya Hospital,Central South University,Changsha 410008;Hunan Center of Drug Safety Evaluation and Research of Drugs,Hunan Key Laboratory of Pharmacodynamics and Safety Evaluation of New Drugs,Changsha 410331;Hunan University of Traditional Chinese Medicine,National Key Laboratory Breeding Base of Chinese Medicine Powders and Innovative Drugs/Technology Innovation Center,Changsha 410208;Cardio-Cerebrovascular Department,Affiliated Union Hospital of Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]中南大学湘雅医院血液科,长沙410008 [2]湖南省药物安全评价研究中心&新药药效与安全性评价重点实验室,长沙410331 [3]湖南省中药粉体与创新药物省部共建国家重点实验室,长沙410208 [4]华中科技大学同济医学院附属协和医院心脑血管科,武汉430030

出　　处：《中南大学学报（医学版）》2019年第12期1330-1337,共8页Journal of Central South University ：Medical Science

基　　金：湖南省科技厅重点研发计划(2018SK2115)~~

摘　　要：目的:研究醒脑静注射液对大鼠全脑缺血性深昏迷的早期保护作用及其机制。方法:雄性SD大鼠采用四血管阻塞法建立全脑缺血性深昏迷模型后分组:模型对照组、醒脑静低、中、高剂量组(1.8,3.6,5.4 mL/kg)、醒脑静(3.6 mL/kg)+PI3K抑制剂组、纳洛酮组(0.04 mL/kg)、醒脑静(3.6 mL/kg)+纳洛酮组(0.04 mL/kg),每组8只。另设8只动物为假手术组。给药组手术后静脉给予不同剂量的醒脑静注射液和/或纳洛酮注射液,给药体积10.0 mL/kg,连续给药3 d;醒脑静+PI3K抑制剂组为造模后立即侧脑室注射10.0μL PI3K抑制剂LY294002(10 mmol/L),然后再给予醒脑静注射液(3.6 mL/kg)。记录首次给药后动物苏醒时间、末次给药后抓力和10 min内自主活动;采用ELISA法检测脑脊液多巴胺(dopamine,DA)和谷氨酸(glutamate,Glu)的含量;采用HE染色观察脑组织病理改变;采用蛋白质免疫印迹法检测海马组织蛋白激酶B(Akt)蛋白及其磷酸化蛋白(p-Akt)和cAMP应答元件结合蛋白(cAMP-response element binding protein,CREB)及其磷酸化蛋白(p-CREB)的表达。结果:与模型对照组比较,醒脑静注射液单次给药能明显缩短大鼠苏醒时间(P<0.05),连续给药3 d能明显增加大鼠抓力、自主活动总路程、活动次数、活动时间(P<0.05或P<0.01),降低DA和Glu的含量(P<0.05或P<0.01),改善海马组织病理改变。醒脑静注射液能显著诱导Akt和CREB蛋白质的磷酸化(P<0.05或P<0.01)。PI3K抑制剂LY294002能明显抑制醒脑静注射液对全脑缺血性深昏迷大鼠上述改善作用(P<0.05或P<0.01)。醒脑静注射液能显著增加纳洛酮注射液对大鼠全脑缺血性深昏迷的上述改善作用(P<0.05或P<0.01)。结论:醒脑静注射液能明显改善大鼠全脑缺血性深昏迷,其机制与上调PI3K/Akt介导的CREB磷酸化,调节神经递质和保护海马组织有关。醒脑静注射液与纳洛酮对大鼠全脑缺血性深昏迷有协同增效作用。Objective:To study the protective effect of Xingnaojing Injection on early global brain ischemia-induced deep coma in rats.Methods:The deep coma model was induced by global brain ischemia by using four-vessel occlusion method in male SD rats.According to the body weight,the rats were randomly divided into 8 groups:a model control group,three different dose of Xingnaojing Injection (1.8,3.6 and 5.4 mL.kg–1) groups,a Xingnaojing Injection (3.6 mL.kg–1) plus PI3K inhibitor group,a naloxone injection (0.04 mL.kg–1) group and a naloxone injection (0.04 mL.kg–1) plus Xingnaojing Injection (3.6 mL.kg–1) group (n=8 per group).In addition,eight animals served as the sham group were performed same operation with the model group excepting no blockage of the blood vessels.After the operation,three different doses of Xingnaojing Injection and/or naloxone injection were given intravenously once a day for three days.Ten μL PI3K inhibitor (LY294002,10 mmol/L) was injected via anterior cerebral ventricle at once after global brain ischemia.The awakening time after the first drug treatment,the grasping power and the autonomous activity within 10 min after the last drug treatment were recorded.The levels of both dopamine (DA) and glutamate (Glu) in cerebrospinal fluid were detected by ELISA.The pathological changes were observed in brain tissue slices with HE staining and the protein levels of Akt/p-Akt and cAMP-response element binding protein (CREB)/p-CREB in hippocampus were detected by Western blotting.Results:Comparing with the model group,single administration of Xingnaojing Injection could significantly shorten the waking time (P<0.05) and continuous administration of Xingnaojing Injection for 3 d could increase grasping power,distance,frequency and duration of autonomous activities (P<0.05 or P<0.01) in the deep coma rat.Also,Xingnaojing Injection could inhibit these increases in neurotransmitters DA and Glu contents (P<0.05 or P<0.01),and improve pathological changes of hippocampal tissue.Xingnaojing Injection s

关 键 词：醒脑静注射液 脑缺血 昏迷 纳洛酮 

分 类 号：R285.5[医药卫生—中药学]