中国组织工程研究2018，Vol.22Issue(13)：2098-2103,6.DOI:10.3969/j.issn.2095-4344.0751
人参皂苷Rg3对体外培养小鼠神经干细胞分化的影响
Effect of ginsenoside Rg3 on mouse neural stem cell differentiation in vitro
摘要
Abstract
BACKGROUND: Application of neural stem cells (NSCs) is of great current interest in neuroscience, but NSCs origin is very limited. And they always differentiate into a large percentage of glial cells and small percentage of neurons in natural differentiation process, so researchers should take effective measures to promote NSCs differentiation into certain offsprings. Previous studies have shown that ginseng saponin ingredients, such as Rb1 and Rg1, have certain influence on NSCs differentiation, but it is unclear whether Rg3 plays a role on NSCs differentiation. OBJECTIVE:To preliminarily investigate the effect of ginsenoside Rg3 on mouse NSCs differentiation into neurons and astrocytes in vitro. METHODS: The fetal cortices of embryonic 14 days (E14) C57BL/6 mice were isolated for culturing primary NSCs. Then passaged NSCs were identified by their purity with NSCs specific antibodies, Nestin and Sox2, by immunofluorescence staining. NSCs were induced for 3 days in the differentiation medium containing ginsenoside Rg3 of different concentrations (blank control, 50 and 250 nmol/L). After that, immunofluorescence staining was used to identify differentiated neurons with neuronal specific antibody, Tuj1, and differentiated astrocytes with astrocyte specific antibody, GFAP. Then, we calculated and statistically analyzed Tuj1+/DAPI and GFAP+/DAPI percentages in the three different groups. Besides, real-time PCR assay was used to test Tuj1 and GFAP mRNA expression in the three groups after 3 days of differentiation. RESULTS AND CONCLUSION: Primary and passaged NSCs were successfully cultured and almost of cells were positive for both Nestin and Sox2, so these high-purity NSCs could be used in the following experiments. Immunofluorescence staining and statistical analysis results showed that compared with the blank control and 250 nmol/L groups, 50 nmol/L group had an obviously increased neuronal percentage after 3 days differentiation (P < 0.01), while the blank control and 250 nmol/L groups had no significant difference (P > 0.05); compared with the blank control group, 50 and 250 nmol/L groups had significantly increased astrocyte percentages (P < 0.05), whereas there was no obvious difference between 50 and 250 nmol/L groups (P > 0.05). The results of real-time PCR assay were similar with the above immunofluorescence results. In conclusion, 50 nmol/L ginsenoside Rg3 can enhance mouse NSCs differentiation into neurons and astrocytes, while 250 nmol/L ginsenoside Rg3 can only promote mouse NSCs differentiation into astrocytes.关键词
人参皂苷Rg3/神经干细胞/分化/神经元/星形胶质细胞/干细胞分类
医药卫生引用本文复制引用
张凤兰,杨璐军,胡炜彦,周姣月,马生霞,肖志成..人参皂苷Rg3对体外培养小鼠神经干细胞分化的影响[J].中国组织工程研究,2018,22(13):2098-2103,6.基金项目
云南省高端人才引进计划(20080A004) （20080A004）
云南省干细胞和再生医学重点实验室(2015DG027)Talent Introduction Program of Yunnan Province,No.20080A004 （2015DG027）
Key Laboratory of Stem Cells and Regenerative Medicine of Yunnan Province,No.2015DG027 （）
