Effects of erythrocytes and serum proteins on lung accumulation of lipoplexes containing cholesterol or DOPE as a helper lipid in the single-pass rat lung perfusion system.
Plasmid DNA-cationic liposome complexes (lipoplexes) accumulate in the lung to a great extent immediately after intravenous administration, and gene expression occurs predominantly in the lung. However, the detailed mechanisms underlying the lung accumulation of lipoplexes are not fully understood. In this study, we investigated the effect of blood components on the lung accumulation of lipoplexes using a single-pass rat lung perfusion system. Two types of lipoplexes, Chol-containing lipoplex ([(32)P]DNA-DOTMA/Chol liposome complex) and DOPE-containing lipoplex ([(32)P]DNA-DOTMA/DOPE liposome complex), pre-incubated with whole blood, serum, or erythrocytes, were injected into the perfused lung via an artery. Similarly to in vivo observations, extensive lung accumulation was observed for both types of lipoplexes after incubation with whole blood during a single passage. The (32)P-labeled lipoplexes pre-incubated with erythrocytes showed similar lung accumulation, whereas their lung accumulation after incubation with serum was significantly reduced, suggesting that erythrocytes would be more responsible blood components for extensive uptake by the perfused lung. However, there was a clear difference in the amounts of the accumulated erythrocytes after intra-arterial injection between the two lipoplex formulations. A significant degree of erythrocyte accumulation was observed when the DOPE-containing lipoplex was injected, whereas the Chol-containing lipoplex failed to induce any significant erythrocyte accumulation in the lung. In vitro experiments showed that the major fraction of both lipoplexes was bound to erythrocytes. These data suggested that Chol-containing lipoplexes bound to erythrocytes before injection dissociate from the erythrocytes and are transferred to the lung capillary endothelial cells during their passage through the lung. In contrast, DOPE-containing lipoplexes bound to erythrocytes cause aggregation and are embolized in the lung capillary with erythrocytes. Thus, the present study demonstrated that the interaction with erythrocytes plays an important role in the lung accumulation of lipoplexes and that neutral helper lipid significantly affects this interaction.
Sakurai
F
F
Department of Drug Delivery Research, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
Nishioka
T
T
Yamashita
F
F
Takakura
Y
Y
Hashida
M
M
D000818:Animals; D001798:Blood Proteins / Q000378:metabolism / Q000502:physiology*; D002784:Cholesterol / Q000493:pharmacokinetics*; D004912:Erythrocytes / Q000378:metabolism / Q000502:physiology*; D020404:Glycerophospholipids / Q000493:pharmacokinetics*; D008081:Liposomes; D008168:Lung / Q000378:metabolism*; D008297:Male; D010477:Perfusion / Q000379:methods; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds / Q000493:pharmacokinetics; D051381:Rats
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D001798:Blood Proteins; D020404:Glycerophospholipids; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; C054005:N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium; D002784:Cholesterol