目的:三七总皂苷可促进去大脑皮质血管后成年大鼠前脑侧脑室室管膜下层神经干细胞Nestin、PCNA和bFGF的表达,以及促进离体胎鼠皮质神经干细胞增殖、分化作用。实验拟采用胶原酶诱导大鼠脑出血模型和免疫组织化学染色法观察三七总皂甙对脑出血大鼠侧脑室室管膜下神经干细胞增殖和分化的作用。 方法:实验于2002/2006年在北京中医药大学神经解剖学实验室完成。①实验材料:Wistar大鼠60只,雄性,清洁级,体质量220—250g,由中国中医研究院实验动物中心提供,将大鼠随机分为正常组、模型组和给药组,20只/组,实验过程中对动物处置符合动物伦理学标准。三七总皂苷血栓通注射液由内蒙古康源药业提供,批准文号:22—5802-内卫药准字(1999)1787号,规格:5mL:175mg。②实验方法:采用胶原酶诱导大鼠脑出血模型,给药组术后腹腔内注射三七总皂苷35mg,1次/d,模型组术后腹腔注射生理盐水,1次/d,正常组不作任何处理。③实验评估:采用免疫组织化学染色法和图像分析技术检测增殖细胞核抗原(PCNA)、β-Ⅲ型微管蛋白(Tuj—1)、胶质原纤维酸性蛋白(GFAP)在室管膜下的表达。 结果:60只大鼠均进入结果分析。脑出血后模型组同侧侧脑室内、外侧壁及背外侧角室管膜下区有增殖细胞核抗原、β-Ⅲ型微管蛋白、胶质原纤维酸性蛋白阳性细胞,给予三七总皂苷干预后,室管膜下细胞增殖细胞核抗原、胶质原纤维酸性蛋白、β-Ⅲ型微管蛋白阳性细胞数目和反应强度较模型组明显增强,差异具有显著性意义(P〈0.05,P〈0.01)。 结论:三七总皂苷具有促进脑出血后侧脑室背外侧角室管膜下区神经干细胞增殖、分化,并向病灶迁移,促进损伤修复的作用。
AIM: Panax notoginseng saponins can promote the expression of Nestin, proliferating cell nuclear antigen and basic fibroblast growth factor in subependymal zone of lateral ventricle in rat brain after cerebral hemorrhage, and accelerate the proliferation and differentiation of cortex neural stem cells of fetal rats. This article investigates the influence of panax notoginseng saponins on proliferation and differentiation of subependymal cells of lateral ventricle in rat brain after intracerebral hemorrhage by the collagenase method and immunohistochemical staining. METHODS: Experiments were performed at the Laboratory of Neuro-anatomy of Beijing University of Chinese Medicine from 2002 to 2006. ①Sixty clean male Wistar rats weighting 220 250 g were provided by Animal Experimental Center of China Academy of Chinese Medical Sciences. Rats were randomly assigned into a normal group, a model group and an administration group, 20 in each group. Animal intervention met the animal ethical standard. Panax notoginseny saponins Xueshuantong injection was offered by Inner Mongolia Kangyuan Yaoye Company, No. 2245802 (1999) 1787, specification 5 mL: 175 mg. ②Rat models of cerebral hemorrhage were established with collagenase as a inductor. Rats in the administration group received 35 mg of panax notoginseny saponins intraperitoneally after the surgery, once a day. Rats in the model group received saline intraperitoneally after the surgery, once a day. Rats in the normal group did not receive any intervention. ③Proliferating cell nuclear antigen, β-Ⅲ tubulin and glial fibrillary acidic protein expression in ependyma was determined by the immunohistochemical staining and image analysis. RESULTS: Sixty rats were involved in the result analysis. Proliferating cell nuclear antigen, β-Ⅲ tubulin and glial fibrillary acidic protein expression were found in subependymal zone of medial wall, lateral wall and the angle of lateral-dorsal of lateral ventricle in rat brain after cerebral hemorrhage in the model