The photosensitiser azure A disrupts mitochondrial bioenergetics through intrinsic and photodynamic effects.

Autor: de Souza BTL; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: byathais@hotmail.com., Klosowski EM; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: eduardomk8@hotmail.com., Mito MS; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: m_s_mito@yahoo.com.br., Constantin RP; Department of Biochemistry, Laboratory of Plant Biochemistry, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: rpconstantin@gmail.com., Mantovanelli GC; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: laine_mantovanelli@hotmail.com., Mewes JM; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: juliana.mewes@outlook.com., Bizerra PFV; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: paulo_veiga22@hotmail.com., da Silva FSI; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: nandasayuri13@gmail.com., Menezes PVMDC; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: pvmcmenezes@hotmail.com., Gilglioni EH; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: gilglioni@hotmail.com., Utsunomiya KS; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: karina.utsunomiya@gmail.com., Marchiosi R; Department of Biochemistry, Laboratory of Plant Biochemistry, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: rmarchiosi@uem.br., Dos Santos WD; Department of Biochemistry, Laboratory of Plant Biochemistry, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: wdsantos@uem.br., Ferrarese-Filho O; Department of Biochemistry, Laboratory of Plant Biochemistry, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: oferrarese@uem.br., Caetano W; Department of Chemistry, Research Nucleus in Photodynamic System, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: wcaetano@uem.br., de Souza Pereira PC; Department of Chemistry, Research Nucleus in Photodynamic System, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: paulocamver@gmail.com., Gonçalves RS; Department of Chemistry, Research Nucleus in Photodynamic System, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: rsgoncalves2@uem.br., Constantin J; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: jconstantin@uem.br., Ishii-Iwamoto EL; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: eliiwamoto@uem.br., Constantin RP; Department of Biochemistry, Laboratory of Biological Oxidations and Laboratory of Experimental Steatosis, State University of Maringá, Maringá, 87020-900, Paraná, Brazil; Department of Biochemistry, Laboratory of Plant Biochemistry, State University of Maringá, Maringá, 87020-900, Paraná, Brazil. Electronic address: rpconstantin@uem.br.
Jazyk: angličtina
Zdroj: Toxicology [Toxicology] 2021 May 15; Vol. 455, pp. 152766. Date of Electronic Publication: 2021 Mar 26.
DOI: 10.1016/j.tox.2021.152766
Abstrakt: Azure A (AA) is a cationic molecule of the class of phenothiazines that has been applied in vitro as a photosensitising agent in photodynamic antimicrobial chemotherapy. It is a di-demethylated analogue of methylene blue (MB), which has been demonstrated to be intrinsically and photodynamically highly active on mitochondrial bioenergetics. However, as far as we know, there are no studies about the photodynamic effects of AA on mammalian mitochondria. Therefore, this investigation aimed to characterise the intrinsic and photodynamic acute effects of AA (0.540 μM) on isolated rat liver mitochondria, isolated hepatocytes, and isolated perfused rat liver. The effects of AA were assessed by evaluating several parameters of mitochondrial bioenergetics, oxidative stress, cell viability, and hepatic energy metabolism. The photodynamic effects of AA were assessed under simulated hypoxic conditions, a suitable way for mimicking the microenvironment of hypoxic solid tumour cells. AA interacted with the mitochondria and, upon photostimulation (10 min of light exposure), produced toxic amounts of reactive oxygen species (ROS), which damaged the organelle, as demonstrated by the high levels of lipid peroxidation and protein carbonylation. The photostimulated AA also depleted the GSH pool, which could compromise the mitochondrial antioxidant defence. Bioenergetically, AA photoinactivated the complexes I, II, and IV of the mitochondrial respiratory chain and the F 1 F O -ATP synthase complex, sharply inhibiting the oxidative phosphorylation. Upon photostimulation (10 min of light exposure), AA reduced the efficiency of mitochondrial energy transduction and oxidatively damaged lipids in isolated hepatocytes but did not decrease the viability of cells. Despite the useful photobiological properties, AA presented noticeable dark toxicity on mitochondrial bioenergetics, functioning predominantly as an uncoupler of oxidative phosphorylation. This harmful effect of AA was evidenced in isolated hepatocytes, in which AA diminished the cellular ATP content. In this case, the cells exhibited signs of cell viability reduction in the presence of high AA concentrations, but only after a long time of incubation (at least 90 min). The impairments on mitochondrial bioenergetics were also clearly manifested in intact perfused rat liver, in which AA diminished the cellular ATP content and stimulated the oxygen uptake. Consequently, gluconeogenesis and ureogenesis were strongly inhibited, whereas glycogenolysis and glycolysis were stimulated. AA also promoted the release of cytosolic and mitochondrial enzymes into the perfusate concomitantly with inhibition of oxygen consumption. In general, the intrinsic and photodynamic effects of AA were similar to those of MB, but AA caused some distinct effects such as the photoinactivation of the complex IV of the mitochondrial respiratory chain and a diminution of the ATP levels in the liver. It is evident that AA has the potential to be used in mitochondria-targeted photodynamic therapy, even under low oxygen concentrations. However, the fact that AA directly disrupts mitochondrial bioenergetics and affects several hepatic pathways that are linked to ATP metabolism, along with its ability to perturb cellular membranes and its little potential to reduce cell viability, could result in significant adverse effects especially in long-term treatments.
(Copyright © 2021 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: