Reis E R, Ferreira L P, Nicola E M D, Borissevitch I
Laser Laboratory of Medicine and Surgery Experimental Center, Faculty of Medical Sciences, State University of Campinas-UNICAMP, PO Box 6111, Campinas, SP, Cep 13083-970, Brazil.
Physics Department, Faculty of Philosophy, Sciences and Letters of Ribeirao Preto, University of Sao Paulo (USP), Ribeirao Preto, Brazil.
Eur Biophys J. 2018 Sep;47(6):601-609. doi: 10.1007/s00249-018-1283-5. Epub 2018 Feb 16.
Protoporphyrin IX (PpIX) is a precursor of heme synthesis and is known to be an active photosensitizer and precursor of photosensitizers applied in photodynamic therapy (PDT) and photodynamic diagnostics (PDD). On irradiation with visible light, PpIX undergoes phototransformation, producing photoproducts which may also be phototoxic and increase its efficacy. The mechanism of PpIX phototransformation depends on environmental characteristics and can be different in vitro and in vivo. In this paper, we present a comparative study of the photoactivity of synthetic PpIX and PpIX extracted from the Harderian gland of ssp Rattus novergicus albinus rats, along with their photoproducts toward murine B16F-10 melanoma cells. It was observed that when irradiated with visible light the endogenous PpIX demonstrates photocytotoxicity ten times higher than the synthetic PpIX. The photoproduct of endogenous PpIX also possesses phototoxicity, though slightly lower than that of PpIX itself. The rate of cell internalization for both endogenous PpIX and its photoproduct was eightfold greater than that obtained for the synthetic porphyrin. This difference might result from a complexation of the native PpIX with some amphiphilic compounds during its synthesis within the Harderian glands, which facilitates the cell uptake of PpIX. Fluorescence microscopy images show that both endogenous and synthetic porphyrins are localized after uptake predominantly in the mitochondrial region of cells.
原卟啉IX(PpIX)是血红素合成的前体,已知是一种活性光敏剂,也是用于光动力疗法(PDT)和光动力诊断(PDD)的光敏剂的前体。在可见光照射下,PpIX会发生光转化,产生的光产物可能也具有光毒性并能提高其疗效。PpIX光转化的机制取决于环境特征,在体外和体内可能有所不同。在本文中,我们对从白化大鼠哈德氏腺中提取的合成PpIX和PpIX及其光产物对小鼠B16F - 10黑色素瘤细胞的光活性进行了比较研究。观察到,在可见光照射下,内源性PpIX表现出比合成PpIX高十倍的光细胞毒性。内源性PpIX的光产物也具有光毒性,尽管略低于PpIX本身。内源性PpIX及其光产物的细胞内化速率比合成卟啉的内化速率高八倍。这种差异可能是由于天然PpIX在哈德氏腺内合成过程中与一些两亲性化合物络合,从而促进了PpIX的细胞摄取。荧光显微镜图像显示,内源性和合成卟啉摄取后主要定位于细胞的线粒体区域。