Characterization of large unilamellar vesicles as models for studies of lipid peroxidation initiated by azocompounds.
The aim of this work was to characterize large unilamellar vesicles (LUVETs) prepared by a hand-driven extrusion device in order to use them for studies of lipid peroxidation and antioxidant activity. Vesicle structure and size were examined by electron microscopy. Lipid and antioxidant content was determined before and after the extrusion procedure. Then LUVETs were subjected to autoxidation initiated by both the lipid-soluble 2,2'-azobis(2,4-dimethylvaleronitrile) and the water-soluble 2,2'-azobis(2-amidinopropane hydrochloride) azocompounds. The results demonstrated that: i) LUVETs prepared with lipid concentrations ranging between 25 and 150 mM were essentially unilamellar and reasonably homogeneous, with an average diameter of 90 nm; ii) the phospholipid, cholesterol and antioxidant amounts retained by filters were about 10-15%; iii) LUVETs were suitable for autoxidation studies initiated by the water-soluble azocompound both in the absence and presence of antioxidants. The lipid-soluble azocompound could be used only at low concentrations and its vesicle content had to be determined since part of the initiator was not incorporated into the lipid bilayer. These data suggest that LUVETs seem to be recommended for studies of lipid peroxidation and antioxidant activity.
Dipartimento di Biochimica G. Moruzzi, Università di Bologna, Italy.
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Fiorentini
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Cipollone
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Galli
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Pugnaloni
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Biagini
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Landi
D000975:Antioxidants / Q000032:analysis; D001391:Azo Compounds / Q000494:pharmacology*; D015227:Lipid Peroxidation; D008563:Membrane Lipids / Q000378:metabolism*; D009570:Nitriles / Q000494:pharmacology*; D010743:Phospholipids / Q000378:metabolism
D000975:Antioxidants; D001391:Azo Compounds; D008563:Membrane Lipids; D009570:Nitriles; D010743:Phospholipids; C055792:2,2'-azobis(2,4-dimethylvaleronitrile)