目的 研究骨碎补(Drynaria fortunei)的化学成分及其对Aβ25-35诱导PC12细胞损伤的保护作用。方法 采用溶剂法提取,利用硅胶、Sephadex LH-20及ODS柱色谱等方法分离纯化,根据质谱(MS),核磁共振法(NMR)等波谱技术鉴定化合物的结构;建立β淀粉样蛋白(Aβ)25-35诱导 PC12细胞损伤体外模型,分别加入不同浓度的化合物,采用四甲基偶氮唑蓝(MTT)法检测细胞活力。结果 从骨碎补中分离并鉴定9个化合物:柚皮苷(1)、新北美圣草苷(2)、5,7-二羟基色原酮-7-O-α-L-鼠李糖基-(1→2)-β-D-葡萄糖基(3)、(E)-4-O-β-D-吡喃葡萄糖基反式咖啡酸(4)、山柰酚(5)、木犀草素(6)、原儿茶酸(7)、补骨脂素(8)和β-谷甾醇(9)。对化合物1~8进行了细胞实验,结果表明,化合物1~8可促进Aβ25-35诱导损伤的PC12细胞增殖,细胞活力随着浓度的增加而加强,差异具有统计学意义(P<0.05)。结论 化合物1~8对Aβ25-35诱导 PC12细胞损伤具有明显的保护作用,为中药骨碎补保护中枢神经功能的主要活性成分。
Abstract
OBJECTIVE To study the chemical constituents from the rhizome of Drynaria fortunei and the protective effects of them on PC12 cells induced by Aβ25-35. METHODS The compounds were isolated and purified by silica gel, Sephadex LH-20, ODS column chromatography, and their structures were identified on basis of spectroscopic methods, such as MS and NMR. PC12 cells were treated with Aβ25-35 to establish the Alzheimer' s disease models. The compounds of different concentrations were added into culture medium to detect the protection. MTT assay was used to detect cell vitality and to observe the protective effects of compounds on PC12 cells induced by Aβ25-35. RESULTS Nine compounds were isolated and identified as naringin(1), neoeriocitrin(2), 5,7-dihydroxychromone-7-neohesperidoside(3), (E)-4-O-β-D-glucopyranosyl caffeic acid(4), kaempferol(5), luteolin(6), protocatechoic acid(7), psoralen(8), and β-sitosterol(9) . The cell experiments were performed on the compounds 1-8 and the results showed they can promote the proliferation of PC12 cells. The cell vitality increase with concentration rising, and the difference is statistically significant (P<0.05). CONCLUSION Compounds 1-8 play an important role in protecting Aβ25-35-induced injury in PC12 cells and they are the main active components of Drynaria fortunei in the protection of central nervous function.
关键词
骨碎补 /
柚皮苷 /
新北美圣草苷 /
结构鉴定 /
Aβ25-35 /
PC12细胞 /
细胞活力
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Key words
Drynaria fortunei /
naringin /
neoeriocitrin /
structural identification /
Aβ25-35 /
PC12 cell /
cell vitality
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中图分类号：
R965
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脚注
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基金
国家自然科学基金面上项目资助(81673581);国家自然科学基金面上项目资助(81673621)
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