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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 protective effect of different extracts of Heterobia live alone on SH-SY5Y cell injury induced by H2O2, and to screen the extraction sites with the strongest pharmacological activity. Methods: The extracts were extracted by reflux, solvent extraction and silica gel column chromatography. h and 250μmol·L-1H2O2 in human neuroblastoma cell line (SH-SY5Y) 24 h, MTT assay cell viability and kit assay intracellular superoxide dismutase (SOD) activity and two (MDA) and the expression of BDNF and NT-3 were detected by immunofluorescence cytochemistry. Results: The extracts of the only live can save the petroleum ether extracts The activity of SH-SY5Y cells induced by H2O2 decreased obviously, and the dose-effect relationship was obvious. All the four extracts could enhance the activity of SOD and decrease the content of MDA. There was no difference in the antioxidant activity between different extracts. The petroleum ether-ethyl acetate group was significantly enhanced BDNF, NT-3 protein expression (94.5% and 97.5%, respectively) compared with the normal group, the neurotrophic effect stronger than the rest of the extract. CONCLUSION: SH-SY5Y cells can be injured by H2O2 to some extent in different parts of the brain, especially in petroleum ether-ethyl acetate group.