藏族药二十五味珍珠丸对缺血性脑卒中大鼠的保护作用及其机制研究  

Protective effect and mechanism of Ershiwuwei Zhenzhu Pills on cerebral apoplexy rats

作　　者：刘智嘉 王存萍 杨斌 王楠 王琴 顾健 谭睿[2] LIU Zhi-jia;WANG Cun-ping;YANG Bin;WANG Nan;WANG Qin;GU Jian;TAN Rui(College of Pharmacy,Southwest Minzu University,Chengdu 610041,China;College of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610031,China)

机构地区：[1]西南民族大学药学院,四川成都610041 [2]西南交通大学生命科学与工程学院,四川成都610031

出　　处：《中国中药杂志》2022年第8期2064-2073,共10页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(82174083);西南民族大学中央高校基本科研业务费专项(2020NZD06);西南民族大学研究生创新型科研项目(CX2020SZ73)。

摘　　要：研究二十五味珍珠丸对缺血性脑卒中大鼠的保护作用。将90只4周龄的SPF级SD雄性大鼠随机平均分成6组(n=15):假手术组、模型组、尼莫地平组(12 mg·kg^(-1))、二十五味珍珠丸高剂量组(400 mg·kg^(-1))、二十五味珍珠丸中剂量组(200 mg·kg^(-1))、二十五味珍珠丸低剂量组(100 mg·kg^(-1))。模型组、尼莫地平组、二十五味珍珠丸组采用改良的线栓法制作永久性大脑中动脉闭塞模型(permanent middle cerebral artery occlusion model,PMCAO),假手术组不插入线栓。造模手术前尼莫地平组和二十五味珍珠丸组每天灌胃给药1次,连续24 d,并于造模前1 h给药1次,假手术组和模型组给予等体积蒸馏水灌胃。测定存活大鼠的神经行为学评分;采用TTC法检测大鼠脑梗死体积;采用HE法观察大鼠脑组织病理学;采用ELISA检测血清中肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素6(IL-6)、丙二醛(MDA)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)表达水平;采用RT-PCR检测大鼠脑组织中Notch 1、Jagged 1、Hes 1、Bcl-2 mRNA的表达;采用Western blot检测大鼠脑组织中凋亡相关因子caspase-3蛋白的表达;采用免疫荧光法检测大鼠脑组织血管内皮生长因子(VEGF)、CD34阳性细胞表达水平。二十五味珍珠丸低、中、高剂量组及尼莫地平组可显著降低永久性大脑中动脉闭塞大鼠神经行为学评分及脑梗死体积,减少神经细胞形态学改变,降低大鼠血清中TNF-α、IL-1β、IL-6表达,提高SOD、CAT活力,降低MDA水平。并且上调Notch 1、Jagged 1、Hes 1及Bcl-2 mRNA表达水平,降低caspase-3蛋白表达水平。同时治疗组中VEGF、CD34阳性细胞数增加,差异均有统计学意义。二十五味珍珠丸对缺血性脑卒中大鼠具有保护作用,其机制可能与抗炎、抗氧化、促进神经细胞增殖、抑制神经细胞凋亡及促进血管新生有关。To study the protective effect of Ershiwuwei Zhenzhu Pills on ischemic stroke rats.Ninety 4-weeks-old SPF male SD rats were randomly divided into 6 groups(n=15):sham operation group,model group,nimodipine group(12 mg·kg^(-1)),Ershiwuwei Zhenzhu Pills high-dose group(400 mg·kg^(-1)),Ershiwuwei Zhenzhu Pills medium-dose group(200 mg·kg^(-1)),Ershiwuwei Zhenzhu Pills low-dose group(100 mg·kg^(-1)).The permanent middle cerebral artery occlusion model(PMCAO)was established in the model group,nimodipine group,and Ershiwuwei Zhenzhu Pills groups by the improved thread plug method,while the sham operation group did not insert the thread plug.Nimodipine group and Ershiwuwei Zhenzhu Pills groups were given intragastric administration once a day for 24 days before the modeling operation,and once 1 hour before the modeling operation,while sham operation group and model group were given equal volumes of distilled water.The neuroethology of the surviving rats was measured;The volume of cerebral infarction in rats was measured by TTC method;The histopathology of rat brain was observed by HE method;The expression levels of tumor necrosis factorα(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),malondialdehyde(MDA),superoxide dismutase(SOD)and catalase(CAT)in serum were detected by ELISA;The mRNA expressions of Notch 1,Jagged 1,Hes 1 and Bcl-2 in rat brain were detected by RT-PCR;Western blot was used to detect the expression levels of caspase-3 protein in rat brain;the expression levels of vascular endothelial growth factor(VEGF)and CD34 positive cells in rat brain were detected by immunofluorescence.The low,medium and high dose groups of Ershiwuwei Zhenzhu Pills and nimodipine group could significantly reduce the neurobehavioral score and cerebral infarction volume of rats with permanent middle cerebral artery occlusion,reduce the morphological changes of nerve cells,decrease the expression of TNF-α,IL-1βand IL-6 in rat serum,increase the activity of SOD and CAT,and reduce the level of MDA.Furthermore,the expression l

关 键 词：二十五味珍珠丸 永久性大脑中动脉闭塞模型 缺血性脑卒中 保护机制 

分 类 号：R29[医药卫生—民族医学]