大枣神经保护作用的活性组分筛选及其作用机制研究 点击下载
论文标题: 大枣神经保护作用的活性组分筛选及其作用机制研究
英文标题:
中文摘要: 目的:筛选大枣神经保护作用的活性组分,并初步阐明其作用机制。方法:以大枣多糖富集组分(1 mg/ml)、大枣多糖去除组分(1 mg/ml)、7种大枣黄酮类成分(儿茶素、原花青素B2、表儿茶素、金丝桃苷、芦丁、槲皮素-3-β-D-葡萄糖苷、山柰酚-3-O-芸香糖苷,记为A、B、C、D、E、F、G,浓度均为3、13、30 μmol/L)和2种大枣核苷类成分(环磷酸腺苷、环磷酸鸟苷,浓度均为3、13、30 μmol/L)分别培养类神经细胞(PC12细胞)24 h后,以叔丁基过氧化氢(tBHP,150 μmol/L)作用于PC12细胞3 h诱导细胞氧化损伤,MTT法检测PC12细胞的存活率。按上述方法培养转染了报告基因质粒(pARE-Luc)的PC12细胞24 h,采用荧光素酶(Luc)法检测各组细胞中抗氧化元件(ARE)的转录水平(以Luc活性反映),以考察各组分抗损伤机制。结果:大枣多糖富集组分能显著提高氧化损伤PC12细胞的存活率,升高转染细胞内ARE转录水平。大枣所含的黄酮类成分中A(30 μmol/L)、B(3~30 μmol/L)和C(10~30 μmol/L)均能显著高细胞存活率,且A(30 μmol/L)、B(3~30 μmol/L)、C(30 μmol/L)、E(30 μmol/L)和F(3~30 μmol/L)能显著提高转染细胞内ARE转录水平。大枣中核苷类成分环磷酸腺苷和环磷酸鸟苷则无明显提高氧化损伤PC12细胞存活率和和转染细胞内ARE转录水平的作用。结论:大枣多糖类成分和大枣黄酮类成分可能是大枣神经保护作用的活性组分,且其作用机制可能与激活ARE转录相关。
英文摘要: OBJECTIVE: To conduct screening for the active ingredients of Ziziphus jujuba with neuroprotective effect and to illuminate their mechanisms of action preliminarily. METHODS: After neuron-like cells (PC12 cells) were respectively cultured in the ingredient of Z. jujuba with polysaccharide enriched (1 mg/ml), that with polysaccharide removed (1 mg/ml), 7 kinds of flavonoid ingredients of Z. jujuba (catechin, procyanidine B2, epicatechin, hyperoside, rutin, quercetin-3-β-D-glucoside and kaempferol-3-O-rutinoside, represented by A, B, C, D, E, F, G, all at the concentrations of 3, 13, 30 μmol/L) and 2 kinds of nucleoside ingredients of Z. jujuba (cyclic adenosine monophosphate and cyclic guanosine monophosphate, both at the concentrations of  3, 13, 30 μmol/L) for 24 h, tert-butyl hydroperoxide (tBHP, 150 μmol/L) was used to act on PC12 cells for 3 h to induce oxidative cellular damage, and MTT assay was employed to detect the survival rate of PC12 cells. The PC12 cells transfected with reporter gene plasmid (pARE-Luc) were cultured as above for 24 h, luciferase (Luc) assay was used to detect the transcriptional levels of the antioxidant response element (ARE) of all groups of cells (reflected as the activity of Luc) so as to investigate the anti-injury mechanism. RESULTS: The ingredient of Z. jujuba with polysaccharide enriched could significantly increase the survival rate of PC12 cells to which oxidative damage was caused and the transcriptional level of ARE in the transfected cells. Among the flavonoid ingredients of Z. jujuba,  A (30 μmol/L), B (3-30 μmol/L) and C (10-30 μmol/L) could significantly increased the survival rate of the cells, and A (30 μmol/L), B (3-30 μmol/L), C (30 μmol/L), E (30 μmol/L) and F (3-30 μmol/L) could obviously increased the activation level of ARE in the transfected cells. However, the nucleoside ingredients of Z. jujuba including cyclic adenosine monophosphate and cyclic guanosine monophosphate had no obvious effect of increasing the survival rate of PC12 cells to which oxidative damage was caused or activating the transcription of ARE in the transfected cells. CONCLUSIONS: The polysaccharide and flavonoid ingredients of Z. jujuba may be the active ingredients which account for the neuroprotective effect against oxidative cellular damage, and their mechanisms of action may be related to the activation of ARE transcription.
期刊: 2016年第27卷第25期
作者: 陈剑平,李中桂,张尚斌,郑平,董婷霞,詹华强
英文作者: CHEN Jianping,LI Zhonggui,ZHANG Shangbin,ZHENG Ping,DONG Tingxia,ZHAN Huaqiang
关键字: 大枣;神经保护;PC12细胞;大枣多糖去除组分;大枣多糖富集组分;抗氧化元件;大枣黄酮;大枣核苷
KEYWORDS: Ziziphus jujuba; Neuroprotection; PC12 cells; Ingredient of Z. jujuba with polysaccharide removed; Ingredient of Z. jujuba with polysaccharide enriched; Antioxidant response element; Flavonoid of Z. jujuba; Nucleoside of Z. jujuba
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