Entamoeba histolytica: production of nitric oxide and in situ activity of NADPH diaphorase in amebic liver abscess of hamsters.

Departamento de Patología Experimental, CINVESTAV-IPN, San Pedro Zacatenco, Mexico.
Parasitology Research (Impact Factor: 2.85). 02/2001; 87(1):49-56. DOI: 10.1007/s004360000287
Source: PubMed

ABSTRACT Entamoeba histolytica trophozoites were inoculated into the liver of hamsters and serum nitrate/nitrite levels [expressed as nitric oxide (NO) production] were determined at different times during amebic liver abscess (ALA) development. We also tested the effects of NO synthase (NOS) inhibitors such as N(G)-nitro-L-arginine methyl ester (L-NAME), aminoguanidine, and dexamethasone during ALA production. Since NOS activity has been correlated with expression of reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) in tissues, we performed histochemistry studies to determine the activity of the latter in livers infected with E. histolytica trophozoites. Production of NO in serum was directly proportional to the size of ALAs, and NOS inhibitors caused low levels of NO and smaller ALAs. Our data suggest that NO does not have any lytic effect on E. histolytica trophozoites and is therefore incapable of providing protection against the amebic liver infection. In addition, NADPHd activity was detected histochemically in hepatocytes and inflammatory cells associated with focal necrosis containing trophozoites. The positive reactivity observed in these parasites may be attributable to a close biochemical similarity of NADPHd to the NADPH:flavin oxidoreductase described in E. histolytica by other investigators.

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    ABSTRACT: The aim of the present study was to determine if the inflammation and/or immunosuppression induced by Entamoeba histolytica may contribute to amebic invasion. Dexamethasone was administered three days before and three days after inoculation of hamsters with E. histolytica. Seven days alter inoculation the animals were sacrificed and the sizes of their amebic liver abscesses were determined. The number of neutrophils, macrophages, T and B cells in the peritoneum as well as the production of nitric oxide and the susceptibility to Listeria monocytogenes infection was also determined. Dexamethasone treatment significantly reduced the number of T lymphocytes in thymus and spleen. The number of neutrophils, macrophages and T lymphocytes in the peritoneal exudate was also reduced as well as the production of nitric oxide and the microbicidal activity against Listeria monocytogenes. However, in the animals treated with a high dose of dexamethasone the size of the liver abscesses was significantly smaller than in the untreated animals. The results suggest that macrophage and T cell-mediated immunity is not relevant as a protective mechanism because tissue invasion by E. histolytica was reduced in immunosuppresed animals. On the contrary, the inflammatory process may contribute to the invasion and liver damage.
    Medical science monitor: international medical journal of experimental and clinical research 10/2004; 10(9):BR317-24. · 1.22 Impact Factor
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    ABSTRACT: NADPH-diaphorase activity has been considered as a nitric oxide synthase (NOS) marker. Therefore, the presence of NADPH-d activity in Entamoeba histolytica suggests that they have NOS activity. The aim of this work was to provide support for this contention. The amebic culture medium or amebic purified proteins induced relaxation of endothelium-denuded rat aortic rings pre-contracted with phenylephrine (10(-6) M), which was inhibited when the amebas were incubated with NG-monomethyl-L-arginine or aminoguanidine (NOS inhibitors), or by pretreatment of the aortic rings with methylene blue. L-Arginine reverted the L-NAME inhibitory effect. In addition, trophozoites produce NO in culture and they have proteins which were recognized by antibodies specific to NOS and show activity of NO synthase. In conclusion, our results provide evidence about the production of NO by trophozoites. This molecule may be responsible for the relaxation elicited by the amebic culture medium and may participate in the pathogenesis of the invasive amebiasis. Index Descriptors and Abbreviations: Entamoeba histolytica; NO, nitric oxide; NOS, nitric oxide synthase; iNOS, inducible nitric oxide synthase; ecNOS, endothelial nitric oxide synthase; NADPH-d, NADPH-diaphorase enzyme; beta-NADPH, beta-nicotinamide-adenine dinucleotide; L-NAME, N-omega-nitro-L-arginine methyl ester hydrochloride; NBT, nitobluetetrazolium; PBS, phosphate-buffered saline; EDTA, ethylenediaminetetraacetic acid; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis
    Experimental Parasitology 01/2003; 104(3-4):87-95. · 2.15 Impact Factor
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    ABSTRACT: Nitric oxide participation during amoebic liver abscess development. Nitric oxide participates in both physiological and pathophysiological functions, and it plays an important role in the mammalian immune system in killing or inhibiting the growth of many pathogens, including parasites, viruses and bacteria. En- tamoeba histolytica is a protozoan parasite that causes amoebiasis, which is characterized by intestinal damage and amoebic liver abscess development. The development of amoebic liver abscess in hamsters is similar to that in humans, whereas mice are resistant to amoebic liver abscess development due to an increase in nitric oxide produc- tion. Unlike in mice, amoebic liver abscess development in hamsters is due to an excess in nitric oxide production or possibly to a greater susceptibility of the hamster to damage caused by nitric oxide. Therefore, it could be important to elucidate if, in humans, an excess in nitric oxide production favors amoebic liver abscess development.
    Medicina 04/2007; 67(2):167-176. · 0.42 Impact Factor

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