摘要
目的:观察具有神经保护作用的银杏叶提取物对海马注射淀粉样β蛋白的阿尔茨海默病模型大鼠神经元凋亡的影响。方法:实验于2002-03/2003-10在河北医科大学第二医院、解放军总医院老年医学研究所、北京大学蛋白质工程国家重点实验室完成。取清洁级成年雌性SD大鼠共30只,随机分为3组,各10只。其中两组大鼠以淀粉样β蛋白25～353μL(4g/L)注射于海马齿状回,10只于当日起给予银杏叶提取物(购自德国威玛舒培博士药厂)2mL腹腔注射,连续7d,为银杏叶提取物组;10只以等量生理盐水腹腔注射7d,为淀粉样β蛋白组;另外10只动物以生理盐水3μL注射于海马齿状回,为对照组;行脱氧核苷酰转移酶法亦即脱氧核糖核酸末端转移酶介导的缺口末端标记和免疫组化染色(Bcl-2和Bax)及双向凝胶电泳观察。结果:30只动物全部进入结果分析,无脱失。①淀粉样β蛋白组及银杏叶提取物组神经元均有凋亡发生。其细胞核被染成棕黄色。典型的凋亡细胞表现为染色质凝集,呈新月形聚集于核膜下。由于切面的不同,凝集的染色质可以不同形态散在于核切面的任意位置,有的以圆形位于核膜下,有的位于核中央,但不象坏死那样边界不清,呈絮状。细胞内有棕黄色粗颗粒分布或棕黄色细腻颗粒弥漫分布者为Bcl-2或bax阳性反应细胞。②淀粉样β蛋白组神经元凋亡率高于银杏叶提取物组(55%,21%);淀粉样β蛋白组Bcl-2反应阳性细胞率低于银杏叶提取物组(15%,47%);淀粉样β蛋白组Bax反应阳性细胞率高于银杏叶提取物组(49%,17%)。③通过分析比较得到的双向凝胶电泳图谱,淀粉样β蛋白组与对照组比较,有26个蛋白质点的体积百分含量发生显著性变化(P<0.05)。而在银杏叶提取物组上述26个发生变化的蛋白点有11个出现回调。结论:银杏叶提取物可使海马注射淀粉样β蛋白组对发生变化的蛋白点回调,对大鼠神经元凋亡有保护作用。
AIM： To investigate the effect of ginkgo biloba extract （GBE） on apeptosis of rat models with Alzheimer disease （AD） induced by amyloid-beta protein （Aβ） intra-hippocampal injection.
METHODS： The experiment was completed in the Second Hospital of Hebei Medical University, Gerontological Institute of General Hospital of Chinese PLA and National Key Laboratory of Protein Engineering of Peking University from March 2002 to October 2003. Thirty adult females rats of clean grade were divided randomly into three groups with 10 in each group. Rats in two groups were injected with 3 μL （4 g/L） Aβ25-35 into hippocampal dentate gyrus. Among them, 10 were injected intravenously with 2 mL GBE （provided by Dr Willmar Schwabe Pharmaceutical Factory, Germany） for 7 days as GBE group; 10 were injected intravenously with the same volume of saline for 7 days as Aβ group; 10 were injected with 3μL saline into hippocampal dentate gyrus as control group. The ratio of cell apoptosis was detected with terminal deoxynucleotidyl transferase （TdT）, i.e. terminal-deoxynucleotidyl transferase mediated nick end labeling （TUNEL）, immunochemical staining （Bcl-2 and Bax） and two dimensional gel electrophoresis. RESULTS： Totally 30 rats entered the final analysis without any loss.① Apeptosis of neurons was found beth in Aβ and GBE groups. The nucleus was buffy, and the typical cell showed coagulation of chromatin and was aggregated under nuclear membrane in crescent shape. Because of various cut, agglutinated chromatin scattered at any site of surface of cut at various shapes. Some of them were located under nuclear membrane in round shape, and some of them were located at center of nucleus with clear border and in catkins shape. It was regarded as positively reacted cell of Bcl-2 or Bax if there were buffy rough granules or buffy smooth granules.②Neuronal apoptosis rate in Aβ group was higher than that in GBE group （55%, 21%）; reactive rate of Bcl-2 positive cells in Aβ group was lower than that in GBE group （15%, 47%）; reactive rate of Bax positive cells in Aβ group was higher than that in GBE group （49%, 17%）. ③ Percentage contents of volume of 26 protein points changed obviously between Aβ group and control group on the basis of two dmensional gel electrophoresis （P 〈 0.05）. However, among 26 protein points, 11 points ameliorated in GBE group.
CONCLUSION： GBE can ameliorate protein points changed by Aβ intrahippecampal injection and protect neuronal apeptosis of rats.
出处
《中国临床康复》
CSCD
北大核心
2006年第11期42-44,i0001,共4页
Chinese Journal of Clinical Rehabilitation
