辽宁碱蓬中磷脂酰胆碱的分离纯化及体外抑癌活性研究
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摘要
碱蓬是一种优质蔬菜、经济作物和具有医疗保健价值的植物资源,目前对碱蓬的开发利用度还比较低,发展前景广阔。
本论文以辽宁碱蓬为材料,采用三氧化二铝柱层析法,优化流动相为95%乙醇:氨水(100:1,V/V)混合洗脱液,获得纯度高于93%、收率高于86%的磷脂酰胆碱(PC)。该方法与单一溶剂洗脱相比,增加了固定相对磷脂的选择性,而且在终产物中无毒性物质残留。
碱蓬源磷脂酰胆碱与其他来源磷脂类产品相比,原料价格低廉、来源广泛,产品中无农药、激素、抗生素类残留等安全问题,原料污染少,无转基因食品隐患,更符合医用高纯PC的要求。
使用纯化所得磷脂酰胆碱对7721、HepG2、MCF-7和Hela四种癌细胞进行体外增殖抑制实验,均表现出较强的抑癌活性,呈时间、剂量依赖方式。抑制作用具有一定的选择性,对肝癌细胞7721和HepG2的抑制率略高于其他两种细胞。制备品与Sigma标准品做平行对照实验,MTT法测定、改良寇氏法计算IC50,证实二者抑制效用相当。
Seepweed is a high-quality vegetables, economic crops and plant resources with the value of health care. There are broad prospects for its development, but at present the development and utilization of seepweed are not enough.
This study is on separation, purification and function analysis of phosphatidylcholine(PC)from Suaeda liaotungensis Kitag.
The phosphatidylcholine, which purity is higher than 93% and the yield higher than 86%, was obtained by using aluminium oxide column chromatography, and optimizing the mobile phase as 95% ethanol: ammonia (100:1, V/V) mixed eluate. Compared with the method of single-eluting solvent, this method increased the selectivity of fixed phase to phospholipid, moreover there are no toxic residue in the final products.
PC from seepweed has more advantages than from other organisms: low price and abundant sources of raw materials, no pesticide residues in products, no hormone residues, no antibiotic residues and other safety problems, less pollution of raw materials, no hidden dangers of transgenic food. It can meet the need for high-purity PC in medicine.
Using the purified phosphatidylcholine on 7721, HepG2, MCF-7 and Hela cells culture,it showed a strong tumor suppressor activity, and a time, dose-dependent manner was found. The inhibition had a certain selectivity. The inhibition rate on 7721 and HepG2 cells were slightly higher than the other two cells. The PC from seepweed and Sigma standard product had the same role on inhibiting cancer cells’growing..
本论文以辽宁碱蓬为材料,采用三氧化二铝柱层析法,优化流动相为95%乙醇:氨水(100:1,V/V)混合洗脱液,获得纯度高于93%、收率高于86%的磷脂酰胆碱(PC)。该方法与单一溶剂洗脱相比,增加了固定相对磷脂的选择性,而且在终产物中无毒性物质残留。
碱蓬源磷脂酰胆碱与其他来源磷脂类产品相比,原料价格低廉、来源广泛,产品中无农药、激素、抗生素类残留等安全问题,原料污染少,无转基因食品隐患,更符合医用高纯PC的要求。
使用纯化所得磷脂酰胆碱对7721、HepG2、MCF-7和Hela四种癌细胞进行体外增殖抑制实验,均表现出较强的抑癌活性,呈时间、剂量依赖方式。抑制作用具有一定的选择性,对肝癌细胞7721和HepG2的抑制率略高于其他两种细胞。制备品与Sigma标准品做平行对照实验,MTT法测定、改良寇氏法计算IC50,证实二者抑制效用相当。
Seepweed is a high-quality vegetables, economic crops and plant resources with the value of health care. There are broad prospects for its development, but at present the development and utilization of seepweed are not enough.
This study is on separation, purification and function analysis of phosphatidylcholine(PC)from Suaeda liaotungensis Kitag.
The phosphatidylcholine, which purity is higher than 93% and the yield higher than 86%, was obtained by using aluminium oxide column chromatography, and optimizing the mobile phase as 95% ethanol: ammonia (100:1, V/V) mixed eluate. Compared with the method of single-eluting solvent, this method increased the selectivity of fixed phase to phospholipid, moreover there are no toxic residue in the final products.
PC from seepweed has more advantages than from other organisms: low price and abundant sources of raw materials, no pesticide residues in products, no hormone residues, no antibiotic residues and other safety problems, less pollution of raw materials, no hidden dangers of transgenic food. It can meet the need for high-purity PC in medicine.
Using the purified phosphatidylcholine on 7721, HepG2, MCF-7 and Hela cells culture,it showed a strong tumor suppressor activity, and a time, dose-dependent manner was found. The inhibition had a certain selectivity. The inhibition rate on 7721 and HepG2 cells were slightly higher than the other two cells. The PC from seepweed and Sigma standard product had the same role on inhibiting cancer cells’growing..
引文
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