目的:从血浆中分离获得低密度脂蛋白(LDL)和进行99m锝 (99mTc)标记，为LDL作为杭癌靶向载体研究作准备。方法:用超速离心法从血桨中分离LDL，从提取分离效果得到较好的梯度分离条件。采用连亚硫酸钠还原法对LDL进行99mTc标记。结果:密度1.006,1.019,1.063的KBr液和1.210的血桨-KBr密度液，以1.5:2.0:3.0:5.0组成不连续密度梯度，经4℃,40 000 r·min-1,24 h超速离心，可一步分离获得电泳纯的LDL。采用连亚硫酸钠还原高得酸进行放射性锝标记，标记率可达39.73%。结论:密度梯度法可一次超速离心提取获得纯度较高的LDL，并可用99mTc标记后用于进一步研究。
Abstract
OBJECTIVE: To purify low density lipoprotein(LDL)from human plasma and label LDL used99m Technetium (99m Tc) for the advanced research of LDL as a carrier targeting antitumor drug to cancer cells.METHODS: A flotated ultracentrifugation of KBr density gradient solution was used to isolate LDL from human plasma.LDL was labeled with 99m Tc using sodium dithionite as a reducing agent of 99m pertechnetate. RESULTS:Ultracentrifugation was performed at 40 000 r·min-1 in a ultracentrifuge for 24 h at 4℃.The ratio of KBr density solutions was 1.5:2:3:5, each density was 1.006,1.019,1.063 and 1.210(serum contained KBr) respectively. The electrophoretic pure LDL was prepared by ultracentrifugation mentioned above. LDL was labeled with 99m Tc. The labeling effectivity was about 39.73%.CONCLUSION: The KBr density gradient ultracentrifugation could be used to isolate and obtain pure LDL from human plasma. And LDL could be labeled with 99m Tc for further research.
关键词
低密度脂蛋白 /
提取 /
锝标记 /
靶向载体
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Key words
low density lipoprotein /
purification /
radiolabeled /
targeting carrier
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参考文献
1 Jackson RL, Morrisett JD, Gotto AM, et al .Lipoproteinstructure and metabolism. Physiol Rev, 1976;56:259.
2 Goldstein JL, Brown MS. Binding and degradation of low density lipoproteins by cultured human fibroblasts. J Biol Chem, 1974;249:5153.
3 Vitols S, Gahrton G, Ost A, et al. Elevated low density lipoprotein receptor activity in leukemic cells with monocytic differentiation. Blood, 1984;63:1186.
4 Rudling MJ, Angelin B, Peterson, et al .Low density lipoprotein receptor activity in human intracranial tumors and its relation to the cholesterol requirement. Cancer Res,1990;50:483.
5 Lundberg B. Preparation of drug-low density lipoprotein complexes for delivery of antitumoral drugs via the low density lipoprotein pathway. CattcerRes, 1987;47:4105.
6 Lestavel-Delattre S, Martin-Nisard F, Clavey V, et al. Lowdensity lipoprotein for delivery of an acrylophenone antineoplastic molecule into malignant cells. Cancer Res,1992;52:3629.
7 Samadi-Baboli G, Favre G, Canal V, et al. Low density lipoprotein for cytotoxic drug targeting: improved activity of elliptinium derivative against B16 melanoma in mice. Br J Cancer, 1993;68:319.
8 江志强，马远鸣.低密度脂蛋白及其作为新型药物载体的研究进展.中国生化药物杂志,1996;17(4):180.
9 Hatch FT, Lees RS. Practical methods for plasma lipoprotein analysis. Adv Lzp Res, 1968;6:1.
10 Lees RS, Garabedian HD, Lees RS, et al:Technetium-99m low density lipoproteins: Preparation and biodis-tribution. J Nucl Med, 1985;26:1056.
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脚注
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基金
国家自然科学基金资助第39670860号
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