Department of Biophysics and Radiation Biology, Semmelweis University, Budapest, Hungary.
Biophysics of Macromolecules, DKFZ, Heidelberg, Germany.
Sci Rep. 2019 Aug 5;9(1):11312. doi: 10.1038/s41598-019-47841-x.
The photodynamic effect requires the simultaneous presence of light, photosensitizer (PS) and molecular oxygen. In this process, the photoinduced damage of cells is caused by reactive oxygen species (ROS). Besides DNA, the other target of ROS is the membranes, separating internal compartments in living cells. Hence, the ability of ROS formation of porphyrins as PSs, in liposomes as simple models of cellular membranes is of outstanding interest. Earlier we compared the binding parameters and locations of mesoporphyrin IX dihydrochloride (MPCl) and mesoporphyrin IX dimethyl ester (MPE), in small unilamellar vesicles (SUV) made from various saturated phosphatidylcholines. In this study, we used the same kinds of samples for comparing the ROS forming ability. Triiodide production from potassium iodide because of light-induced ROS in the presence of molybdate catalyst was applied, and the amount of product was quantitatively followed by optical spectrometry. Furthermore, we demonstrated and carefully studied SUVs disruption as direct evidence of membrane destruction by the methods of dynamic light scattering (DLS) and fluorescence correlation spectroscopy (FCS), applying unsaturated phosphatidylcholines as membrane components. Although the ROS forming ability is more pronounced in the case of MPCl, we found that the measured disruption was more effective in the samples containing MPE.
光动力作用需要光、光敏剂(PS)和分子氧同时存在。在这个过程中,细胞的光诱导损伤是由活性氧(ROS)引起的。除了 DNA,ROS 的另一个靶标是细胞膜,它将活细胞的内部隔室分隔开来。因此,作为 PS 的卟啉形成 ROS 的能力,以及在脂质体中作为细胞膜的简单模型,是非常有趣的。我们之前比较了中卟啉 IX 二盐酸盐(MPCl)和中卟啉 IX 二甲酯(MPE)在各种饱和磷脂酰胆碱制成的小单层囊泡(SUV)中的结合参数和位置。在这项研究中,我们使用了相同的样品来比较 ROS 的形成能力。应用含有钼酸盐催化剂的光诱导 ROS 从碘化钾中产生三碘化物,通过光学光谱法定量跟踪产物的数量。此外,我们应用不饱和磷脂酰胆碱作为膜成分,通过动态光散射(DLS)和荧光相关光谱(FCS)等方法,证明并仔细研究了 SUV 破坏,作为膜破坏的直接证据。虽然 MPCl 中 ROS 的形成能力更为明显,但我们发现,在含有 MPE 的样品中,测量到的破坏更为有效。