NITRIC OXIDE-MEDIATED APOPTOSIS IN RAT MACROPHAGES SUBJECTED
TO SHIGA TOXIN FROM ESCHERICHIA COLI
JL Baronetti1, N Angel Villegas1, V Aiassa1, ME Suárez2, MG Paraje1, I Albesa1
1 Higiene y Microbiología. Departamento de Farmacia, Facultad de Ciencias
Químicas, Universidad Nacional de Córdoba, Argentina. 2 Laboratorio de
Microbiología del Hospital de Niños de la Santísima Trinidad de Córdoba., Argentina
Purpose
The aim of this study was to investigate the participation of nitric oxide (NO) in the
apoptosis of rat peritoneal macrophages induced by culture supernatants and shiga
toxin from Escherichia coli.
Material and methods Shiga toxin (Stx) was purified from a clinically isolated E. coli O157:H7 stain using a
receptor-mediated affinity chromatography. Purity of toxin was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining. Rat peritoneal macrophages, obtained by Percoll gradient, were exposed to different dilutions of E. coli cell free supernatants or Stx for 24 hs at 37°C in a 5% CO2
humidified atmosphere. After incubation, apoptotic cells (propidium iodide staining) and NO production (Griess reaction) were evaluated. In some experiments, the cultures were performed in the presence of inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine (AG, 1mM). Results Peritoneal macrophages incubated in presence of E. coli supernatants showed an
increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. humidified atmosphere. After incubation, apoptotic cells (propidium iodide staining) and NO production (Griess reaction) were evaluated. In some experiments, the cultures were performed in the presence of inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine (AG, 1mM). Results Peritoneal macrophages incubated in presence of E. coli supernatants showed an
increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. receptor-mediated affinity chromatography. Purity of toxin was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining. Rat peritoneal macrophages, obtained by Percoll gradient, were exposed to different dilutions of E. coli cell free supernatants or Stx for 24 hs at 37°C in a 5% CO2
humidified atmosphere. After incubation, apoptotic cells (propidium iodide staining) and NO production (Griess reaction) were evaluated. In some experiments, the cultures were performed in the presence of inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine (AG, 1mM). Results Peritoneal macrophages incubated in presence of E. coli supernatants showed an
increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. humidified atmosphere. After incubation, apoptotic cells (propidium iodide staining) and NO production (Griess reaction) were evaluated. In some experiments, the cultures were performed in the presence of inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine (AG, 1mM). Results Peritoneal macrophages incubated in presence of E. coli supernatants showed an
increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli.
Material and methods
Shiga toxin (Stx) was purified from a clinically isolated E. coli O157:H7 stain using a
receptor-mediated affinity chromatography. Purity of toxin was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining. Rat peritoneal macrophages, obtained by Percoll gradient, were exposed to different dilutions of E. coli cell free supernatants or Stx for 24 hs at 37°C in a 5% CO2
humidified atmosphere. After incubation, apoptotic cells (propidium iodide staining) and NO production (Griess reaction) were evaluated. In some experiments, the cultures were performed in the presence of inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine (AG, 1mM). Results Peritoneal macrophages incubated in presence of E. coli supernatants showed an
increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. humidified atmosphere. After incubation, apoptotic cells (propidium iodide staining) and NO production (Griess reaction) were evaluated. In some experiments, the cultures were performed in the presence of inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine (AG, 1mM). Results Peritoneal macrophages incubated in presence of E. coli supernatants showed an
increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli.
receptor-mediated affinity chromatography. Purity of toxin was assessed by sodium
dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining.
Rat peritoneal macrophages, obtained by Percoll gradient, were exposed to different
dilutions of E. coli cell free supernatants or Stx for 24 hs at 37°C in a 5% CO2
humidified atmosphere. After incubation, apoptotic cells (propidium iodide staining) and NO production (Griess reaction) were evaluated. In some experiments, the cultures were performed in the presence of inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine (AG, 1mM). Results Peritoneal macrophages incubated in presence of E. coli supernatants showed an
increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli.
humidified atmosphere. After incubation, apoptotic cells (propidium iodide staining)
and NO production (Griess reaction) were evaluated. In some experiments, the
cultures were performed in the presence of inducible nitric oxide synthase (iNOS)
inhibitor aminoguanidine (AG, 1mM).
Results
Peritoneal macrophages incubated in presence of E. coli supernatants showed an
increment in apoptosis levels as well as in the NO production. Furthermore, the inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated with a diminution in the percentage of apoptotic cells in these cultures, indicating the participation of this metabolite in the apoptotic process. On the other hand, in order to identify one of the probable products involved in these phenomena, presents in the culture supernatants, rat peritoneal macrophages were cultivated in presence of different concentrations of Stx. In similar form at observed with the culture supernatants, the treatment of cells with Stx induced an increment in the NO production and apoptotic levels. In addition, these parameters also were reverted by the aggregated of AG at cultures. Conclusion Different E. coli-related products have been shown to induce apoptosis in a variety of
cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. cells types. However, the precise relationship between NO synthesis and induction of apoptosis has not been intensely investigated. In this study, we demonstrated that the treatment with E. coli supernatants, or with the major virulence factors of this
pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli. pathogen, Stx, induces nitric oxide-mediated apoptosis of rat peritoneal macrophages. These results could contribute to better understand of the immunopathology of E. coli.
increment in apoptosis levels as well as in the NO production. Furthermore, the
inhibition of NO synthesis, induced by the addition of AG at cultures, was correlated
with a diminution in the percentage of apoptotic cells in these cultures, indicating the
participation of this metabolite in the apoptotic process. On the other hand, in order
to identify one of the probable products involved in these phenomena, presents in
the culture supernatants, rat peritoneal macrophages were cultivated in presence of
different concentrations of Stx. In similar form at obs
