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大脑缺血缺氧的损伤,导致体内神经干细胞巢微环境结构和功能的紊乱,实验室前期研究表明三七总皂苷、人参总皂苷能够促进中风后神经干细胞增殖、迁移和分化,促进脑缺血后缺氧后VEGF的表达。黄芪甲苷、麦冬皂苷是否能够通过作用于神经干细胞巢内的星形胶质细胞和脑微血管内皮细胞,通过调控VEGF的表达促进神经干细胞的增殖和分化尚不清楚。目的:分别将胎大鼠星形胶质细胞、脑微血管内皮细胞与神经干细胞共培养,模拟体内复杂的神经干细胞巢微环境,研究黄芪甲苷、麦冬皂苷是否能够通过作用于神经干细胞巢内的星形胶质细胞和脑微血管内皮细胞,通过调控VEGF的表达促进神经干细胞的增殖和分化,启动脑损伤结构和功能的修复。材料:取孕15 d SD大鼠,用以分离培养神经干细胞、星形胶质细胞、和微血管内皮细胞。方法:利用Transwell装置,分别将大鼠星形胶质细胞、微血管内皮细胞和神经干细胞共培养。设立空白对照组、模型组和中药有效成分组,于氧糖剥夺模型4小时后2h、4h和6h3个时间点分别收集处理细胞和培养液,做ELISA和免疫荧光双标染色。结果:大鼠星形胶质细胞、脑微血管内皮细胞分别与神经干细胞共培养条件下,与空白对照组比较,黄芪甲苷、麦冬皂苷可显著增加Brdu、Tuj-1和Vimentin阳性细胞数(P<0.05),显著上调VEGF的表达(P<0.05)。结论:黄芪甲苷、麦冬皂苷通过作用于大鼠星形胶质细胞和微血管内皮细胞,促进神经干细胞的增殖和分化,可能与黄芪甲苷、麦冬皂苷通过促进VEGF的分泌调控神经发生有关。
Cerebral ischemia and hypoxia damage, leading to the structure and function of the neural stem cell nests microenvironment in vivo disorders, laboratory studies have shown that notoginseng total saponins, ginseng total saponins can promote stroke after stroke neural stem cells proliferation, migration and differentiation, and promote cerebral ischemia Post-hypoxia VEGF expression. Astragaloside and Ophiopogon japonicas could promote the proliferation and differentiation of neural stem cells by regulating the expression of VEGF through acting on astrocytes and brain microvascular endothelial cells in the nests of neural stem cells. OBJECTIVE: To co-culture fetal rat astrocytes and cerebral microvascular endothelial cells with neural stem cells respectively to simulate the complex microenvironment of neural stem cells in vivo and to investigate whether Astragaloside and Ophiopogon japonicus could act on the neural stem cell nests Of astrocytes and brain microvascular endothelial cells, promote the proliferation and differentiation of neural stem cells by regulating the expression of VEGF, and initiate the repair of brain injury structure and function. MATERIALS: 15-day-old SD rats were used to isolate and culture neural stem cells, astrocytes and microvascular endothelial cells. METHODS: Rat astrocytes, microvascular endothelial cells and neural stem cells were co-cultured with Transwell apparatus respectively. The blank control group, the model group and the traditional Chinese medicine active ingredient group were set up. The cells and media were collected at 4h, 2h, 4h and 6h after oxygen deprived model, respectively. ELISA and immunofluorescence double staining were performed. Results: Astragaloside and Radix Ophiopogon japonicus could significantly increase the number of Brdu, Tuj-1 and Vimentin-positive cells (P <0.01) after co-culture of rat astrocytes and cerebral microvascular endothelial cells with neural stem cells P <0.05), significantly upregulated the expression of VEGF (P <0.05). Conclusion: Astragaloside and Radix Ophiopogon japonicus can promote the proliferation and differentiation of neural stem cells through acting on astrocytes and microvascular endothelial cells in rats, which may be related to the effects of astragaloside IV and Ophiopogon japonicus saponins on promoting the secretion of VEGF .