Protective effect of gypenosides on primarily cultured fetal rat mesencepimlic dopaminergic neurons
10.3760/cma.j.issn.1671-8925.2009.11.007
- VernacularTitle:绞股蓝皂苷对原代培养胚鼠中脑多巴胺能神经细胞的保护作用
- Author:
Dong XIONG
1
;
Wei-Xin LI
;
Peng WANG
;
Dong JIA
;
Hao YANG
;
Guo-Dong GAO
Author Information
1. 第四军医大学唐都医院
- Keywords:
Gypenosides;
Dopaminergic neurons;
Tyrosine hydroxylase;
Inducible nitric oxide synthase;
Cell apoptosis
- From:
Chinese Journal of Neuromedicine
2009;8(11):1106-1110
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the protective effect of gypenosides (GPs) on primarily cultured mesencephalic dopaminergic (DA) neurons in embryonic rats. Methods Ventral mesencephalic neurons were primarily cultured in the presence of GPs at the concentrations of 100, 200, 400, and 800 μg/ml, with culture medium as the blank control and nerve growth factor (NGF) as the positive control. At 6, 12, 24, 48, 72 and 96 h, the cells were observed under inverted microscope and the cell viability was assessed using MTT assay to optimize the concentration of GPs and culture time. The expression of tyrosine hydroxylase (TH)-positive neurons in the 3 groups was detected by immunocytochemical staining. In the cells with or without GPs pretreatment, 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) was added, and the cell apoptosis was detected by flow cytomertry and the expression of inducible nitric oxide synthase (iNOS) observed by immunocytochemical staining. Results MTT assay showed that incubation of the cells with 400 μg/mL GPs for 48 h resulted in the highest cell viability, which was uncomparable with that cell incubated in the presence of NGF (P>0.05). Immunocytochemistry demonstrated a significant increase in TH-positive neurons in GPs and NGF groups as compared with those in the blank control group (P<0.05). In the cells with 400 μg/ml GPs pretreatment, iNOS expression and cell apoptosis rates, as shown by immunocytochemistry and flow cytomertry, were significantly higher than those in the blank control group, but is still obviously lower than those in MPTP-treated cells without GPs pretreatment (P<0.05). Conclusion GPs may offer protective effect on primarily cultured DA neurons from embryonic rats.
