To describe the action mechanisms of Bacteriochlorin a (BCA), a second generation photosensitizer, in phosphate buffer (PB) and in dimyristoyl phosphatidylcholine (DMPC) liposomes we carried out oxygen consumption and ESR measurements. In PB, where BCA was in a monomer-dimer equilibrium, our results suggested that the oxygen consumption was related to the BCA monomers concentration in solution. Incorporation of BCA in DMPC liposomes, by promoting the monomerization of BCA, increased 9-fold the oxygen consumption in comparison to the value in PB. The use of specific singlet oxygen quenchers (Azide and 9,10-Anthracenedipropionic acid) in ESR and oxygen consumption experiments allowed us to assert that BCA was mainly a type II sensitizer when it was incorporated in DMPC. Finally, the cell survival of WiDr cells after a PDT treatment was measured for cells incubated with BCA in cell culture medium and cells incubated with BCA in DMPC. Irrespective of the dye concentration, the cell survival was lower when liposomes were used. This effect could be the result of a better BCA monomerization and/or a different BCA uptake in cells.

译文

为了描述第二代光敏剂细菌氯蛋白a (BCA) 在磷酸盐缓冲液 (PB) 和二肉豆蔻酰磷脂酰胆碱 (DMPC) 脂质体中的作用机理,我们进行了耗氧量和ESR测量。在PB中,BCA处于单体-二聚体平衡状态,我们的结果表明,耗氧量与溶液中BCA单体的浓度有关。通过促进BCA的单体化,将BCA掺入DMPC脂质体中,与PB相比,氧消耗增加了9倍。在ESR和耗氧实验中使用特定的单线态氧猝剂 (叠氮化物和9,10-蒽二丙酸) 使我们可以断言,当BCA掺入DMPC中时,BCA主要是II型敏化剂。最后,对于在细胞培养基中与BCA孵育的细胞和在DMPC中与BCA孵育的细胞,测量PDT处理后WiDr细胞的细胞存活率。无论染料浓度如何,使用脂质体时细胞存活率较低。这种效果可能是更好的BCA单体化和/或细胞中不同的BCA摄取的结果。

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