脑心安胶囊对慢性脑缺血致血管性认知障碍大鼠胶质细胞激活及炎性反应的调节作用  

Regulation of Naoxin’an Capsule on Glial Cell Activation and Inflammatory Response in Rats with Chronic Cerebral Hypoperfusion-induced Vascular Cognitive Impairment

作　　者：韩广卉 李东岳 余虹霓 甄伟哲 马涛[1] HAN Guang-hui;LI Dong-yue;YU Hong-ni;ZHEN Wei-zhe;MA Tao(Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China;School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学东方医院,北京100078 [2]北京中医药大学中医学院,北京100029

出　　处：《中国实验方剂学杂志》2021年第19期46-55,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81973786,81673929)。

摘　　要：目的:以慢性脑缺血致血管性认知障碍(VCI)大鼠为模型,探讨脑心安胶囊对VCI大鼠脑内胶质细胞激活及炎性损伤的保护作用。方法:150只大鼠随机分为假手术组(20只)和造模组(130只),采用双侧颈动脉结扎法(2-VO)造模。造模成功的87只大鼠,随机分为模型组、阳性药组(安理申,0.5 mg·kg-1),脑心安低、中、高剂量组(0.18,0.36,0.72 g·kg-1),每组17~18只大鼠。经灌胃给予脑心安8周后,采用Morris水迷宫和避暗箱实验检测脑心安对VCI大鼠记忆能力的影响,苏木素-伊红(HE)染色法观察大鼠大脑海马CA1区神经细胞结构变化,原位末端标记法(TUNEL)染色检测大鼠海马神经细胞凋亡水平,蛋白免疫印迹法(Western blot)检测大鼠脑内胶质纤维酸性蛋白(GFAP),离子钙结合衔接分子-1(Iba-1)表达水平,测定p38有丝分裂原活化蛋白激酶(p38 MAPK)和核转录因子-κB(NF-κB)磷酸化水平,酶联免疫吸附测定法(ELISA)测定脑内炎性因子白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平。结果:与假手术组比较,模型组大鼠空间学习记忆能力和记忆保持能力均明显下降(P<0.05,P<0.01),大脑海马CA1区神经元出现明显损伤,细胞凋亡率显著升高(P<0.01),GFAP,Iba-1表达均显著上调(P<0.01),p38 MAPK,NF-κB磷酸化水平显著增加(P<0.01),炎性因子IL-1β,TNF-α水平升高(P<0.01)。与模型组比较,脑心安各剂量组均能显著改善模型大鼠空间学习记忆能力和记忆保持能力(P<0.05,P<0.01),改善海马CA1区神经元损伤,降低神经细胞凋亡率(P<0.05,P<0.01),下调GFAP,Iba-1表达量(P<0.01),降低p38 MAPK,NF-κB磷酸化水平(P<0.05,P<0.01),降低TNF-α,IL-1β水平(P<0.01)。结论:脑心安胶囊通过抑制慢性脑缺血诱发的大鼠脑内小胶质细胞和星形胶质细胞活化,抑制p38 MAPK和NF-κB激活导致的中枢神经炎性损伤,改善慢性脑缺血导致的VCI。Objective:To investigate the protective effect of Naoxin’an capsule(NC)against glial cell activation and inflammatory damage in brain of rats with chronic cerebral hypoperfusion-induced vascular cognitive impairment(VCI). Method: One hundred and fifty rats were randomly divided into a sham operation group(n=20)and a modeling group(n=130). Following the modeling with the two vessels occlusion(2-VO)technique,87 successfully modeled rats were randomly divided into the model group,positive drug group(aricept,0.5 mg·kg-1),and low-,medium-,and high-dose(0.18,0.36,0.72 g·kg-1)NC groups,with 17-18 rats in each group. After intragastric administration of NC for eight weeks,the Morris water maze test and passive avoidance test were conducted to detect the effects of NC on learning and memory ability of VCI rats.Changes in neuronal structure of rat hippocampal CA1 area were observed by hematoxylin-eosin(HE)staining,and the neuronal apoptosis in hippocampus by terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)staining. Western blot assay was used to detect the expression levels of glial fibrillary acidic protein(GFAP),ionized calcium-binding adapter molecule 1(Iba-1),phosphorylated p38 mitogenactivated protein kinase(p38 MAPK),and phosphorylated nuclear factor κB(NF-κB),followed by the measurement of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the brain by enzyme-linked immunosorbent assay(ELISA). Result: Compared with the sham operation group,the model group displayed obviously decreased spatial learning and memory ability and memory retention ability(P<0.05,P<0.01),neuronal damage in hippocampal CA1 area,enhanced neuronal apoptosis(P<0.01),up-regulated GFAP and Iba-1(P<0.01),elevated phosphorylation of p38 MAPK and NF-κB(P<0.01),and increased IL-1β and TNF-α(P<0.01). Compared with the model group,NC at each dose significantly improved the spatial learning and memory ability and memory retention ability of VCI rats(P<0.05,P<0.01),ameliorated the neuronal damage in hi

关 键 词：慢性脑缺血 血管性认知障碍 脑心安胶囊 胶质细胞 炎性反应 

分 类 号：R2-0[医药卫生—中医学] R22