摘要
目的:通过比较独活不同提取物对H2O2致SH-SY5Y细胞损伤的保护作用,筛选药理活性最强的提取部位方法:以回流提取,溶剂萃取及硅胶柱层析方法得到独活不同提取部位预保护6 h后与250μmol·L-1H2O2共同作用于人神经母细胞瘤细胞株(SH-SY5Y)24 h,MTT检测细胞活力,并试剂盒法测定细胞内超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,以及免疫荧光细胞化学法检测脑源性神经营养因子(BDNF)和神经因子-3(NT-3)的表达量结果:独活各提取物除石油醚萃取物外均能救护H2O2所致SH-SY5Y细胞活力降低,且有明显的量效关系;4种提取物均能增强SOD活性,降低MDA含量,不同提取部位抗氧化作用无差异;石油醚-乙酸乙酯组显著增强BDNF、NT-3蛋白表达(与正常组比较,分别为94.5%和97.5%),其神经营养作用强于其余提取物。结论:独活不同提取部位均可不同程度保护H2O2损伤SH-SY5Y细胞,其中石油醚-乙酸乙酯组作用最强。
Objective： To compare the neuroprotective effect of different extracts of Angelica pubescens on SH-SY5Y cells injury induced by hydrogen peroxide （H2O2） so as to screen the most pharmacologically active extracts. Method： Different extracts were obtained by means of reflux extraction, solvents extraction or silica gel column chromatography. Pro-incubation of different extracts on SH-SY5Y cells for 6 h prior to co-treat with 250 μmol·L-1 H2O2 for 24 h, the cell viability were observed by MTT assay. Meanwhile, the activity of superoxide dismutase （SOD） and content of malondialdehyde （MDA） was determined.The expression of brain-derived neurotrophic factor （BDNF） and nerve factor-3 （NT-3） by immunofluorescence was measured. Result： All extracts showed the protective effect against decrease of cell viability induced by H2O2 and there was dose-effect relationship except the extract of petrdeum ether（PE） These extracts enhanced SOD activity and lowered H2O2-induced MDA release. Therefor,they exhibited the similar effect of anti-oxidation;According to increasing the level of BDNF and NT-3（94.5% and 97.5% vs. control, relatively）, we found that the PE-EtOAc extract was better neuroprotective than others. Conclusion： The different extracts of A. pubescens show some protective effect on H2O2 induced SH-SY5Y cells injury and the PE-EtOAc extract has the most pharmacologically activity.
出处
《中国实验方剂学杂志》
CAS
北大核心
2013年第24期184-188,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81173580)
辽宁省自然科学基金项目(201102144)
沈阳市科技专项资金项目(F11-264-1-42)
关键词
独活
SH-SY5Y细胞
H2O2
抗氧化
神经营养
Angelica pubescens
SH-SY5Y cells
hydrogen peroxide
anti-oxidation
neuroprotective
