Fresh aromatic herbs containing methylchavicol did not exhibit the pro-oxidative effects of pure methylchavicol on a human hepatoma cell line, HepG2.

Methylchavicol (CH(3)-CV), an important aromatic constituent of different plants like tarragon and basils, has been shown to be carcinogenic by a mechanism yet unclear, although it has been reported that carcinogenicity of CH(3)-CV in rodent might be linked to its metabolic conversion into a genotoxic electrophilic metabolite generated through a two steps bioactivation pathway catalyzed by cytochrome P450 enzymes and sulfotransferases. The induction of carcinogenesis by certain agents has been associated with the generation of oxidative stress. The aim of the present study was to determine whether pure methylchavicol applied on a human hepatoma cell line, HepG2, could promote oxidative stress and might alter the expression of procarcinogenic biomarkers such as the drug-metabolizing enzyme (CYP2E1), the inducible form of nitric oxide synthase (iNOS) and might induce the expression of Cu/Zn-superoxide dismutase (Cu/Zn-SOD) and Mn-SOD that control the redox equilibrium of the cells. CH(3)-CV was shown to cause a significant induction of oxidative stress, as revealed by luminol-dependent chemiluminescence (LDCL) and to alter dramatically the expression of CYP2E1, iNOS and Mn-SOD, indicating that the toxic effect of CH(3)-CV could be mediated through a nitric oxide dependent mechanism. Under similar experimental conditions, the extracts from tarragon, chervil and basil did not induce such biological changes. These results provide evidence that the generation of an oxidative stress may be a significant event occurring during CH(3)-CV-induced toxicity. It also suggests that natural extracts containing different amounts of CH(3)-CV (tarragon, chervil and basil) did not elicit such toxicity and might contain compounds able to counteract this detrimental property.

Mast cells as a source and target for nitric oxide.

Mast cells (MC), which are tissue-resident cells found widely distributed in the body, are derived from primitive hematopoietic cells. MC produce a variety of biologically active substances such as histamine, proteases, lipid derivatives and numerous cytokines and chemokines in response to immunologic or non-immunologic stimuli. Of interest, it has been reported that rodent MC can also be a source of nitric oxide (NO) derivatives, that they synthesize spontaneously, or only after activation, depending on their subtype. This synthesis appears to be under the control of the expression of the inducible isoform of the nitric oxide synthase (iNOS) and of the constitutive neuronal NOS (nNOS). MC might thus be able to influence the survival and functions of other types of NO-sensitive cells in close vicinity. Apart from being a source of NO, MC can also be the target for NO and its derivatives. Indeed, survival and reactivity of rodent MC is influenced by NO derivatives produced by MC themselves or by other cellular elements in close contact with the MC in tissues. By contrast, the existence of such mechanisms of cross-talk between MC and NO remains poorly documented in humans. If evidence are supplied in favor of such relationship, pharmacological modulation by agents acting at the level of the NO pathway might be of interest in order to regulate the functions of MC in immunologic, neoplastic, inflammatory and other conditions.

[Taurine: a particular aminoacid with multiple functions]. / La taurine: un aminoacide particulier aux fonctions multiples.

Taurine has multiple biological and metabolic functions: this particular B-amino acid is an antioxidant, it conjugates biliary acids, detoxifies some xenobiotics and modulates intracellular calcium levels. Moreover, taurine plays an important part in osmoregulation, neuromodulation and stabilization of the membranes. Taurine is looked upon as an "essential amino acid" in some particular situations associating inadequate intake or synthesis and major loss of biliary salts. Clinically, taurine has been used with varying degrees of success in the treatment of several pathologies (cardiovascular diseases, cystic fibrosis, alcoholism, retinal degeneration, hepatic disorders). Being found in the secretions of the mammalian reproductive tract, it preserves the motility of the spermatozoa, supports their capacitation, improves the chances of success of fertilization and the early embryonic development. This is why it can be found in some culture media for in vitro fertilization.

Inhibition of mouse mast cell proliferation and proinflammatory mediator release by benzodiazepines.

Mast cell (MC) activation may occur in vitro and in vivo following stimulation with various immunologic or nonimmunologic agents. Such activation leads to the release of several biological mediators, including vasoactive amines, nitric oxide and cytokines, which account for the adverse effects observed during allergic reactions. While high affinity binding sites for benzodiazepines (BZDs) have been reported on MC, the effects of the ligation of these receptors on the proliferation of, and the mediator release from, these cells are poorly documented. In the present work, we have examined the effects of midazolam and of diazepam on the proliferation of mucosal (MMC)-like and of serosal (CTMC)-like mouse MC. In addition, we have studied the effects of these BZDs on beta-hexosaminidase, TNF-alpha and nitrite release induced from mouse mast cells through IgE receptor activation. We demonstrated that each of the two BZDs studied inhibited the proliferation of MMC- and CTMC-like elements in a dose-dependent fashion (10 to 100 microM). Furthermore, the BZDs inhibited the IgE-mediated release of beta-hexosaminidase, TNF-alpha and nitrites from MMC- or CTMC-like cells. Altogether, these data provide new insights into the pharmacological regulation of MC activation and may lead to the discovery of new and potent antiallergic compounds.

Involvement of cyclic nucleotides in the immunomodulatory effects of nitric oxide on murine mast cells.

Nitric oxide (NO) is synthesized by various cells involved in inflammatory reactions and may then act on mast cells. In the present work, we attempted to clarify the role of this molecule on the proliferation and IgE-mediated activation of mouse bone-marrow-derived mast cells obtained by culture in the presence of IL-3 (BMMC). Treatment of BMMC with increasing concentrations of sodium nitroprusside (SNP) induced a dose-dependent inhibition of 3H-thymidine incorporation (IC50 = 50 microM) without affecting cell survival under 100 microM. Furthermore, nitric oxide dramatically decreased beta-hexosaminidase and TNF-alpha release induced by Fc epsilon RI ligation on BMMC (respectively 45 and 57% for 100 microM). These inhibitory effects are mediated at least in part through enhancement of intracellular cyclic nucleotides levels since: 1) intracellular cGMP and cAMP levels increased within minutes after NO treatment, 2) treatment of BMMC with a cAMP analogue induced antiproliferative effect on BMMC and 3) pretreatment of BMMC with a cAMP antagonist partly reversed the inhibitory activity of SNP.

Nitric oxide pathway is induced by Fc epsilon RI and up-regulated by stem cell factor in mouse mast cells.

Murine stem cell factor (SCF) induces the differentiation of mucosal mast cells (MMC) into connective tissue mast cells (CTMC) and potentiates mediator release induced by aggregation of high-affinity IgE receptors (Fc epsilon RI). In the present work, we investigated the effect of Fc epsilon RI aggregation on nitric oxide (NO) pathway induction in the different subsets of mast cells, as well as the contribution of SCF in this induction. Inducible NO synthase (iNOs) expression was not evidenced in non-stimulated MMC obtained by culture of hematopoietic progenitors in the presence of interleukin-3, whereas IgE-antigen-stimulated MMC expressed iNOs mRNA and protein and synthesized nitrites. Long-term treatment of MMC with SCF, allowing them to differentiate into CTMC, induced iNOs expression in non-stimulated cells and up-regulated iNOs expression and generation of NO derivatives induced by IgE-antigen stimulation. Thus, NO derivatives generated by mast cells could participate in inflammatory reactions during allergic stimulation.

Evidence for direct interaction between mast cells and Leishmania parasites.

When stimulated through IgE-(or IgG-) immune complexes with parasite antigens, mast cells can release several cytokines, including IL-4, IL-6, IL-10, IL-12, Interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) that may influence the host response to Leishmania major in modulating lesion size and persistence during experimental infection in the mouse. Moreover, recent data demonstrated that mast cells are able to be antibody-independently activated by direct contact with bacteria, making them important elements in innate immunity. Given these data, we asked whether cell-parasite contact could directly induce mast cell mediator release and whether mast cells could be infected by L. major or L. infantum parasites. In this study, we showed that a pure homogeneous population of mouse bone marrow derived mast cells (BMMC) in contact with living L. major or L. infantum promastigotes, but not with attenuated parasites or soluble parasite antigens, released preformed mediators such as beta-hexosaminidase and the preformed pool of TNF-alpha within minutes. Furthermore, direct cell-parasite contact induced TNF-alpha synthesis by mast cells within hours. Moreover, we demonstrated by in vitro co-culture experiments that metacyclic L. major or L. infantum promastigotes are directly infective for a significant proportion of BMMC and are transformed into intracellular amastigotes. Taken together, these data suggest that mast cell can participate in the first line of defence, i.e. innate immunity, during local cutaneous infection with Leishmania parasites.