RESUMO
Photodynamic therapy (PDT) is a modality of therapy that involves the activation of photosensitive substances and the generation of cytotoxic oxygen species and free radicals to promote the selective destruction of target tissues. This study analyzed the application of PDT to Tritrichomonas foetus, a scourged and etiological agent of bovine trichomoniasis, a sexually transmitted infectious disease. As it is an amitochondrial and aerotolerant protozoan, it produces energy under low O2 tension via hydrogenosome. T. foetus from an axenic culture was incubated with photosensitizer tetrasulfonated aluminium phthalocyanine and then irradiated with a laser source (InGaAIP) at a density of 4.5Jcm-2. The DNA integrity of the control and treated group parasites was analyzed by conventional gel electrophoresis and comet assay techniques. In previous results, morphological changes characterized by apoptotic cell death were observed after T. foetus was submitted to PDT treatment. In the treated groups, T. foetus DNA showed a higher concentration of small fragments, about 200pb, in gel electrophoresis after PDT. In the comet assay, the DNA tail percentage was significantly higher in the treated groups. These results demonstrate that PDT leads to DNA fragmentation with changes in nuclear morphology and apoptotic features.
Assuntos
Bovinos/parasitologia , Dano ao DNA/genética , DNA de Protozoário/genética , DNA de Protozoário/efeitos da radiação , Fotoquimioterapia/métodos , Tritrichomonas foetus/efeitos dos fármacos , Tritrichomonas foetus/genética , Animais , DNA de Protozoário/efeitos dos fármacos , Relação Dose-Resposta a Droga , Relação Dose-Resposta à Radiação , Doses de Radiação , Tritrichomonas foetus/efeitos da radiaçãoRESUMO
Entamoeba histolytica, the protozoan responsible for human amoebiasis, exhibits a great genome plasticity that is probably related to homologous recombination events. It contains the RAD52 epistasis group genes, including Ehrad51 and Ehrad54, and the Ehblm gene, which are key homologous recombination factors in other organisms. Ehrad51 and Ehrad54 genes are differentially transcribed in trophozoites when DNA double-strand breaks are induced by ultraviolet-C irradiation. Moreover, the EhRAD51 recombinase is overexpressed at 30 min in the nucleus. Here, we extend our analysis of the homologous recombination mechanism in E. histolytica by studying EhRAD51, EhRAD54, and EhBLM expression in response to DNA damage. Bioinformatic analyses show that EhRAD54 has the molecular features of homologous proteins, indicating that it may have similar functions. Western blot assays evidence the differential expression of EhRAD51, EhRAD54, and EhBLM at different times after DNA damage, suggesting their potential roles in the different steps of homologous recombination in this protozoan.