“白脉散”抗脑缺血ECG对实验性MCAO/R模型神经保护作用的研究

Reasearch on the Neuroprotective Effects of Anti-stroke Effective Compound Group of BaiMai Powder on Experimental MCAO/R Model

目的研究民族大复方"白脉散"中抗脑缺血有效成分组(ECG)对大鼠脑缺血再灌注损伤的保护作用及相关机制。方法 60只雄性SD大鼠随机分为假手术组、模型组、银杏叶提取物组、"白脉散"ECG高、中、低剂量组,采用线栓法制作大鼠大脑中动脉闭塞再灌注模型,缺血2 h再灌注,再灌注24 h后进行神经功能评分和提高躯体摆动实验(EBST),然后连续灌胃给药7 d,1次/d。利用常规HE染色法观察病灶皮层细胞的病理变化,TTC染色法观察脑梗死体积,并检测病灶区脑匀浆上清中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量、还原型谷胱甘肽含量(GSH)、一氧化氮合酶(NOS)活性。结果与模型组比较,"白脉散"ECG高、中、低3个剂量组神经功能评分和对侧旋转次数水平显著改善(P<0.05、P<0.01或P<0.001);病理学研究显示病灶处皮质神经元肿胀坏死有所减轻;TTC染色结果显示脑梗塞体积显著减少;脑匀浆上清SOD活性和GSH含量显著升高,MDA含量显著降低(P<0.01或P<0.001),总NOS(T-NOS)、诱导型NOS(iNOS)、构建型NOS(cNOS)活性均显著降低(P<0.05、P<0.01或P<0.001)。结论 "白脉散"ECG可保护MCAO/R模型大鼠神经功能,其机制可能与"白脉散"抗脑缺血ECG的清除自由基,增强机体抗氧化能力,减轻脑缺血再灌注所致的氧化性损伤,减轻脑梗塞体积及抑制NOS尤其是iNOS介导的NO生成有关,为在"有效成分组"理论指导下攻克民族大复方物质基础研究难题提供依据。

Objective To study the neuroprotective effects and related mechanisms of effective compound group（ECG） of national great prescription ＂ BaiMai Powder＂ on cerebral ischemia - reperfusion injury in rats. Methods 60 adult Sprague Dawley rats were randomly divided into 6 groups ： sham - operated group, cerebral isehemie reperfusion group, extract of Ginkgo Biloba group,＂ BaiMai Powder＂ ECG（ high, moderate,low dose groups）. Then a rat cerebral ischemia/reperfusion injury was made through embo- lization of the middle cerebral artery, the neurological score and elevated body swing test（EBST） were taken after 2 h MCAO ,fol- lowed by 24 h Of reperfusion, then all groups were administered intragastrieally for 7d, respectively. The brains were removed to observe the morphological change of cortical ceils through hematoxylin -eosin staining （HE） , cerebral infarction volume by 2,3,5 chlorinated three phenyl paclobutrazol four nitrogen （TTC）, and then detected the activities superoxide dismutase （SOD） and nitric oxide synthase（NOS） and contents of malondialdehyde （MDA） and glutathione （GSH） in brain homogenate. Results Compared with MCAO/R group, the neurological score and time of contralateral rotation of treatment ECG were decreased, and the difference was statistically significant（ P 〈 0.05 ,P 〈 0.01or P 〈 0. 001 ）. Pathological studies showed that swelling and necrosis of cortical neurons in lesions has lessened, cerebral infarction volume in ECG was decreased compared with MCAO/R group, GSH content and SOD activity significantly increased（ P 〈 0.01 or P 〈 0. 001 ）, and MDA content and NOS activity were significantly decreased （ P 〈 0.05, P 〈 0.01 or P 〈 0.001 ）. Conclusion ECG can remarkably improve the neural behavioral functions of model rats and reduce the pathological changes by scavenging oxygen radicals, improving the antioxidation capacity of brain tissue, reducing oxida- tion injury caused by cerebral ischemia reperfusion, reducing cerebral infarction volume and inhibiting the generation of NO mediated by NOS, especially iNOS, which will provide the basis for conquering the problems in studying the physical basis of national great prescription under the guidance of theory of ECG.