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Bacteroides fragilis supernatant Extracts Enriched in Phenylacetic Acid Induce a Cytotoxic Effect in Mammalian Cells

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Advances in Microbiology 2015年第10期5卷 730-736页

作者： Laís s.Falcao Eduardo N.F.Antunes Eliane O.Ferreira Heidi Pauer Maria Teresa V.Romanos Rossiane C.Vommaro sérgio H.seabra Daniela s.alviano celuta s.alviano Antonio Jorge R.da silva Leandro A.Lobo Regina Maria C.P.Domingues Departamento de Ciências Básicas Pólo Universitário de Nova FriburgoUniversidade Federal FluminenseNova FriburgoBrasil Instituto de Microbiologia Prof.Paulo de Góes Universidade Federal do Rio de JaneiroRio de JaneiroBrasil Instituto de Biofísica Carlos Chagas Filho Universidade Federal do Rio de JaneiroRio de JaneiroBrasil Centro Universitário Estadual da Zona Oeste—UEZO Rio de JaneiroBrasil Instituto de Pesquisa Produtos Naturais Universidade Federal do Rio de JaneiroRio de JaneiroBrasil

Bacteroides species are nearly half of the fecal flora community and some are host symbionts crucial to host nutrition and systemic immunity. Among Bacteroides species B. fragilis strains are considered to be the opportunistic ones, being the most isolated anaerobic bacteria in clinical samples. Cell-free supernatants of 65 B. fragilis strains were assayed and they were capable of inducing vacuolating phenotype on Vero cells lineage. The supernatant of the Bacteroides fragilis ATCC 23745 strain was elicited to have the strongest vacuolating effect on Vero cells monolayers and peritoneal macrophages. some drastic cell alterations were observed, such as a general disorganization of cytoplasm and chromatin condensation, evidencing cell death. By transmission electron microscopy it was confirmed that the vacuoles observed were, in fact, swollen mitochondria. An immunocytochemical assay, TUNEL, was used to confirm this hypothesis and showed that Vero cells and peritoneal macrophages were dying by apoptotic process after exposition of B. fragilis cell-free supernatant. Physical analysis of the apoptotic factor has revealed properties similar to short-chain fatty acids. After gas chromatography and mass spectrometry analysis, phenylacetic acid (PA) was characterized as the major compound present in the most purified active fraction. We believe that the PA is responsible for the pro-apoptotic effect elicited by the supernatant of B. fragilis cultures.

关键词： Bacteroides fragilis Vacuolization Apoptosis Vero cells Lineage Peritoneal Macrophages Phenylacetic Acid

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