| Autor: |
Kitano, Soichi, Higashimoto, Yuichiro, Harada, Shoji, Sano, Masaru, Kurata, Tsunehiko, Yamaguchi, Yu, Kunitomo, Masaru, Haginaka, Jun, Yamagishi, Sho-ichi |
| Předmět: |
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| Zdroj: |
Annals of Clinical Biochemistry; Sep2010, Vol. 47 Issue 5, p460-466, 7p, 2 Diagrams, 3 Charts, 2 Graphs |
| Abstrakt: |
Background: Circulating oxidized low-density lipoproteins (LDLs) (ox-LDLs) could be a sensitive marker to predict future cardiovascular events. However, a method to evaluate oxidized forms of LDLs systemically in human plasma is not yet established. In this study, we developed a novel and convenient high-performance liquid chromatography (HPLC) method for measuring ox-LDL levels in humans. Methods: Human plasma lipoproteins were separated by a modified HPLC method using a diethylaminoethyl-type anionexchange gel column with stepwise elution. Ox-LDLs were detected by postcolumn reaction with a regent containing cholesterol esterase and cholesterol oxidase. Particle size of each LDL fraction separated by HPLC was determined in 61 healthy subjects. Results: Our HPLC method separated LDLs into three fractions, which were designated as LDL-1, LDL-2 and LDL-3, on the basis of their negative charges, with LDL-3 the most strongly retained fraction migrating fastest in the anodic direction, a property that reflects the net negative charge of the molecule. Western blot analysis revealed that apolipoprotein B100 in LDL-3 fraction was the most fragmented and oxidatively modified. When LDLs were oxidized in vitro by Cu2+ or 2,2-azobis (2-aminopropane)-2HCl or modified by various aldehydes, all of the LDL fractions migrated at the position of LDL-3. Further, among three fractions, particle size was smallest in LDL-3 fraction. Conclusion: Here, we developed a convenient HPLC method and identified LDL-3 as oxidized LDL fractions, although ox-LDLs were present in LDL-2 fraction, albeit lesser concentrations than in LDL-3 subfraction. Measuring ox-LDL levels in human plasma by this method may be useful to evaluate atherosclerotic disorders. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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