Cigarette smoke condensates from all cigarettes tested were found to be potent inducers of AHH enzyme in cultured human lymphocytes and, with the exception of Kent Lights and Carlton CSC's, all were found to be toxic under the experiment conditions. Most of the AHH inducing activity was found in basic and neutral fractions of the lAl standard cigarettes. A radiometric assay of BP metabolites in cultured human lymphocytes was developed in which we were able to separate the primary metabolites and the secondary metabolites from the parent compound (BP) by neutral alumnia HPLC. The primary metabolites were further separated by a selective enzyme hydrolysis and/or reverse phase HPLC.

Cigarette smoke contributes to many diseases. The actions of second and third hand smoke, which have implications for non-smokers and the very young, are just beginning to be appreciated. The overarching hypothesis of this project is that cigarette smoke has different injurious actions on alveolar cells based on chronological age. The purpose here was to learn more about the susceptibility of alveolar cells to cigarette smoke extract (CSE)- induced injury by performing studies on pulmonary alveolar and endothelial cells derived from neonatal, young, and old rats. The aims involved: 1. Developing cell culture models to study age-related effects of cigarette smoke on alveolar type I cells and microvascular endothelial cells from the lung, and 2. Using these models to examine the effects of CSE on markers of oxidative stress, inflammation and aging in alveolar cells harvested from neonatal, young and old rats. Descriptive and experimental studies involved using a variety of cell culture, biochemical and molecular techniques, including gene expression arrays. The most significant findings were that: 1. primary proliferating alveolar type I cells were used to develop novel cell culture model systems, including single culture, co-culture and three-dimensional cultures that were used to study the effects of CSE; 2. Hydrogen peroxide production by endothelial cells was markedly reduced by co-culturing with AT I cells; 3. Gene expression profiling of oxidative stress-specific pathways suggest that genes responsible for both stopping production of H2O2 or mopping-up H2O2 are involved; and 4. Cigarette smoke shortens telomeres of cells from neonates, but unexpectedly preserves telomere length of cells from young and old rats. Data from telomeric pathway-specific gene expression arrays suggest that there are age-related differences in response to gene expression to CSE. The significant conclusions are: 1. Contrary to prior observations, alveolar type I cells demonstrate prolonged proliferative capacity; 2. Alveolar type I cells likely play an important role in ameliorating CSE-induced oxidative stress; and 3. Neonatal alveolar cells may be more susceptible to the deleterious effects of CSE including telomere shortening. These novel model systems and observations provide new ways to study cigarette smoke-associated lung dysfunction.

Introduction: Asthma is a common debilitating disease of the airways that afflicts an estimated 300 million worldwide, causing reduction in physical activity, lost school/work days, and even death. There are many known and suspected risk factors of asthma; however, there is much controversy over prior and current cigarette smoking. Approximately 25% of the United States population currently smokes, with a quarter of these being asthma patients. Another 22 to 43 percent of asthmatics are ex-smokers. Objectives: (1) To estimate the prevalence for lifetime asthma in the adult US population; (2) to determine prevalence odds ratios (POR) of lifetime asthma based on questionnaire (smoking status, tobacco consumption) after adjustment of potential confounding variables; (3) to determine POR of lifetime asthma based on laboratory values (serum cotinine); and (4) to assess the validity of self-reported measures (smoking status and tobacco consumption) using serum cotinine as the gold standard.Methods: The National Health and Nutrition Examination Survey (NHANES) 1999-2002 is a proportional cross-sectional sample that uses weights to be representative. Crude odds ratios were obtained through univariate analysis; multiple logistic regression analysis was used to obtain adjusted odds ratios of asthma. Interactions for age, gender, and race/ethnicity were explored. Validity measures included sensitivity and specificity tests for self-reported smoking and non-parametric correlation of tobacco consumption with serum cotinine levels.Results: The overall prevalence of lifetime asthma among n=10,252 adults was 11.56% (95%CI 10.45-12.66). Analyses were stratified by race/ethnicity due to significant interaction. After adjusting for age, gender, body mass index, and family history of asthma, ex-smoking non-Hispanic Whites, non-Hispanic Blacks, and other races had odds ratios of 1.57 (95%CI 1.26-1.97), 1.52 (95%CI 1.01-2.27), and 1.97 (95%CI 1.01-3.83), respectively, relative to never smokers within their respective race/ethnic groups. Sample persons with a family history of asthma and increasing body mass index were significant predictors for lifetime asthma among all race/ethnic groups. Based on laboratory values, non-Hispanic White respondents with serum cotinine levels of 0.011 to Discussion: Self-reported smoking and tobacco consumption are valid measures of tobacco use. The present study found no significant relationship between current smoking and lifetime asthma. Despite the limited findings, asthmatic smokers make up a distinct, difficult-to-treat subgroup for which future treatment research should address.

O objetivo deste estudo foi avaliar a estabilidade de cor e rugosidade de superfície de 3 compósitos odontológicos com partículas diferentes: nanohíbrido (Tetric N-Ceram - Ivoclar Vivadent), híbrido (Z250 - 3M ESPE) e microhíbrido com matriz silorano (Filtek P90 3M - ESPE), submetidos à fumaça de cigarro. Para isso, foram obtidos 60 corpos-de-prova (8mm de diâmetro X 2mm de espessura), 20 para cada tipo de material restaurador utilizado, dos quais 10 receberam polimento com lixas dágua em abrasividades decrescentes (Grupo1) e a outra metade não foi submetida a nenhum tipo de acabamento/polimento ficando sob efeito apenas de uma matriz de poliéster (Grupo 2). Após armazenamento dos corpos de prova em água destilada a 37°C por 24 horas, foram realizadas as primeiras leituras de cor (Colorímetro Vita Easy Shade) na escala de cor CIEL*a*b* e rugosidade superficial (Rugosímetro Surfcorder SE 1700 - Kosakalab). Em seguida, as amostras foram expostas à fumaça de 20 cigarros por um período de 10 minutos cada, (Marlboro - Philip Morris), sendo que entre um cigarro e outro as amostras foram submetidas à escovação em dispositivo padronizado e lavagem em água corrente. Após ação do número total de cigarros, foram realizadas leituras finais de cor e rugosidade. Os valores de &Delta;E, &Delta;L, &Delta;a e &Delta;b foram analisados estatisticamente utilizando 2 way ANOVA e Teste de Bonferroni (p<0.05) e os valores de rugosidade média (Ra) foram analisados isoladamente para cada tipo de material (teste t Student, p < 0.05). Verificou-se que o compósito Tetric N apresentou maior alteração de cor, em níveis clinicamente inaceitáveis (&Delta;E>3,3) e de rugosidade de superfície em relação aos demais compósitos quando utilizada a tira de poliéster como acabamento. Concluiu-se que a ausência de polimento aumenta a capacidade manchadora do cigarro sobre todos os compósitos e a rugosidade de superfície dos materiais, com exceção de P90. / The purpose of this study was to evaluate the color stability and surface roughness of three dental composites with different type of fillers: Nanohybrid (Tetric N-Ceram), Hybrid (Filtek Z250-3M ESPE) and Microhybrid silorane based composite (Filtek P90- 3M ESPE), exposed to cigarette smoke. 20 specimens (8mm diameter X 2mm thickness) of each material were prepared and separated into two groups (n=10) according to the surface treatment : Group 1- polymerization through a polyester film strip and polishing with water sandpaper in decreasing abrasiveness (600, 800 e 1200); and Group 2- polymerization through a polyester film strip, without any polishing. After immersion in destilled water 37° for 24 hours, baseline color of all specimens was measured using reflectance spectrophotometer (Easy Shade - Vita) with CIEL*a*b* system, surface roughness was also measured using a rugosimeter (SJ-201P Mitutoyo, Tokyo, Japan). After base line measurements, the specimens were exposed to smoke from 20 cigarettes in 10 minutes each, (Marlboro - Philip Morris), between one cigarette and other, the specimens were submitted to brushing in a standardized device and washed with water. After the action of the total number of cigarettes, final measurements of color and surface roughness were performed. Values of &Delta;E, &Delta;L, &Delta;a e &Delta;b were statistically analyzed using 2 way ANOVA e Teste de Bonferroni (p<0.05) and values of average surface roughness (Ra) were analyzed separately for each type of material (teste t Student, p < 0.05). It was found that the composite Tetric N-Ceram showed increased of surface roughness and greater color change in clinically unacceptable levels (&Delta;E>3,3), in comparison with the other composites when used the polyester strip without polishing. It was concluded that the absence of polishing increases the cigarette staining and surface roughness on all composites except for Filtek P90.

O objetivo deste estudo foi avaliar a estabilidade de cor e rugosidade de superfície de 3 compósitos odontológicos com partículas diferentes: nanohíbrido (Tetric N-Ceram - Ivoclar Vivadent), híbrido (Z250 - 3M ESPE) e microhíbrido com matriz silorano (Filtek P90 3M - ESPE), submetidos à fumaça de cigarro. Para isso, foram obtidos 60 corpos-de-prova (8mm de diâmetro X 2mm de espessura), 20 para cada tipo de material restaurador utilizado, dos quais 10 receberam polimento com lixas dágua em abrasividades decrescentes (Grupo1) e a outra metade não foi submetida a nenhum tipo de acabamento/polimento ficando sob efeito apenas de uma matriz de poliéster (Grupo 2). Após armazenamento dos corpos de prova em água destilada a 37°C por 24 horas, foram realizadas as primeiras leituras de cor (Colorímetro Vita Easy Shade) na escala de cor CIEL*a*b* e rugosidade superficial (Rugosímetro Surfcorder SE 1700 - Kosakalab). Em seguida, as amostras foram expostas à fumaça de 20 cigarros por um período de 10 minutos cada, (Marlboro - Philip Morris), sendo que entre um cigarro e outro as amostras foram submetidas à escovação em dispositivo padronizado e lavagem em água corrente. Após ação do número total de cigarros, foram realizadas leituras finais de cor e rugosidade. Os valores de &Delta;E, &Delta;L, &Delta;a e &Delta;b foram analisados estatisticamente utilizando 2 way ANOVA e Teste de Bonferroni (p<0.05) e os valores de rugosidade média (Ra) foram analisados isoladamente para cada tipo de material (teste t Student, p < 0.05). Verificou-se que o compósito Tetric N apresentou maior alteração de cor, em níveis clinicamente inaceitáveis (&Delta;E>3,3) e de rugosidade de superfície em relação aos demais compósitos quando utilizada a tira de poliéster como acabamento. Concluiu-se que a ausência de polimento aumenta a capacidade manchadora do cigarro sobre todos os compósitos e a rugosidade de superfície dos materiais, com exceção de P90. / The purpose of this study was to evaluate the color stability and surface roughness of three dental composites with different type of fillers: Nanohybrid (Tetric N-Ceram), Hybrid (Filtek Z250-3M ESPE) and Microhybrid silorane based composite (Filtek P90- 3M ESPE), exposed to cigarette smoke. 20 specimens (8mm diameter X 2mm thickness) of each material were prepared and separated into two groups (n=10) according to the surface treatment : Group 1- polymerization through a polyester film strip and polishing with water sandpaper in decreasing abrasiveness (600, 800 e 1200); and Group 2- polymerization through a polyester film strip, without any polishing. After immersion in destilled water 37° for 24 hours, baseline color of all specimens was measured using reflectance spectrophotometer (Easy Shade - Vita) with CIEL*a*b* system, surface roughness was also measured using a rugosimeter (SJ-201P Mitutoyo, Tokyo, Japan). After base line measurements, the specimens were exposed to smoke from 20 cigarettes in 10 minutes each, (Marlboro - Philip Morris), between one cigarette and other, the specimens were submitted to brushing in a standardized device and washed with water. After the action of the total number of cigarettes, final measurements of color and surface roughness were performed. Values of &Delta;E, &Delta;L, &Delta;a e &Delta;b were statistically analyzed using 2 way ANOVA e Teste de Bonferroni (p<0.05) and values of average surface roughness (Ra) were analyzed separately for each type of material (teste t Student, p < 0.05). It was found that the composite Tetric N-Ceram showed increased of surface roughness and greater color change in clinically unacceptable levels (&Delta;E>3,3), in comparison with the other composites when used the polyester strip without polishing. It was concluded that the absence of polishing increases the cigarette staining and surface roughness on all composites except for Filtek P90.

The FHIT gene is located on the most fragile site in the human genome. FHIT gene deletions are among the earliest and most frequent events in carcinogenesis, particularly in carcinogen-exposed tissue. Previous work in mouse and cell culture models established FHIT to be an authentic tumor suppressor. Re-expression of FHIT in cell culture causes cell death via initiation of apoptosis, but the precise mechanism underlying this process is unclear. It is well established that cellular transition from normal to transformed occurs in multiple steps and requires the accumulation of several genetic changes. Relying on the compelling phenotype of tumor development in FHIT knockout mice, this project aimed to elucidate a mechanism through which FHIT-deficient cells are primed to survive multiple genetic and environmental stresses, and promote progression of cancer. My work indicates that FHIT expression is required for the normal cellular response to oxidative stress, and presents evidence that in the absence of FHIT, an oxidative stress response pathway is superinduced. When FHIT is depleted from cells exposed to cigarette smoke, the expression of a subset of oxidative stress response genes is enhanced. Enhanced activation of these genes can occur as an adapative response to stress induced by reactive oxygen species production, and is frequently detected in cancer. Investigation into the mechanism underlying the enhanced gene expression determined that FHIT loss is associated with decreased levels of the transcriptional repressor Bach1. In this manner, we propose that loss of Fhit supports an antioxidant program that is pivotal in establishing and maintaining carcinogenic transformation.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Inflammatory and degenerative pathophysiological processes within the central nervous system (CNS) and platelets are important causes of human disease promoted by cigarette smoke. The objective of the present study was to evaluate activity of the enzymes that hydrolyze nucleotides and nucleosides in synaptosomes of cerebral cortex and platelets from rats exposed to aged and sidestream diluted smoke of commercial cigarettes. The animals were randomly divided into two groups (ten rats in each group): control (not exposed to cigarette smoke) and smoke (exposed to cigarette smoke). The smoke group was exposed via inhalation through the nose only 15 minutes per day inside a cigarette smoke exposure chamber during 3 weeks. The results demonstrated that E-NTPDase (with ATP and ADP as substrate) activity was significantly higher in the smoke group (p<0.05), while the 5'- nucleotidase with AMP as substrate demonstrated decreased activity in the smoke group (p<0.05). However, in platelets activities demonstrated that E-NPP and ENTPDase with ATP as substrate and 5´-nucleotidase with AMP as substrate were significantly higher in the smoke group (p<0.05) as compared to the control group, while E-NTPDase with ADP as substrate demonstrated decreased activity in the smoke group (p<0.05). A decrease of adenosine, a neuroprotective nucleoside, was observed in brain, which could lead to damage of the brain tissue. However, in platelets, there was an increase of adenosine, which may reflect adaptative changes. These alterations caused by cigarette smoke on ectonucleotidase activities may assist in verifying pathophysiological effects in biological systems. / Os processos patofisiológicos inflamatórios e degenerativos no sistema nervoso central (SNC) e plaquetas são importantes causas de doenças humanas promovidas pela fumaça de cigarro. O objetivo do presente estudo foi avaliar a atividade das enzimas que hidrolisam nucleotídeos e nucleosídeos em sinaptossomas de córtex cerebral e plaquetas de ratos expostos à fumaça envelhecida e diluída inalada por via secundária de cigarros comerciais. Os animais foram aleatoriamente divididos em dois grupos (10 ratos por grupo): controle (não exposto a fumaça de cigarro) e fumo (exposto a fumaça de cigarro). O grupo fumo foi exposto à fumaça de cigarro 15 minutos por dia no interior de uma câmara de exposição durante 3 semanas. Em relação ao sinaptossoma de córtex cerebral os resultados demonstraram que a atividade da E-NTPDase (ATP e ADP como substrato) aumentou significativamente no grupo exposto à fumaça de cigarro (p<0.05) e a atividade da 5'-nucleotidase (AMP com substrato) diminuiu neste grupo (p<0.05), comparado ao grupo controle. Nas plaquetas foi demonstrado um aumento na atividade das enzimas E-NPP e ENTPDase (ATP como substrato) e 5 -nucleotidase (AMP como substrato) no grupo exposto à fumaça de cigarro (p<0.05) quando comparado ao grupo controle. Já a ENTPDase (ADP como substrato) demonstrou uma diminuição de sua atividade no grupo exposto à fumaça de cigarro (p<0.05). Pode-se sugerir que a provável diminuição da adenosina, uma molécula neuroprotetora, pode levar ao dano no tecido do encéfalo. Entretanto, em plaquetas, houve um aumento da adenosina, a qual pode refletir mudanças adaptativas. As alterações na atividade das ectonucleotidases em animais submetidos à exposição de fumaça de cigarro podem contribuir na elucidação de seus efeitos patológicos em sistemas biológicos.

Current smoking cessation guidelines recommend nicotine replacement therapy (NRT) to assist pregnant smokers to quit, but this is without strong evidence for effectiveness and safety. Nicotine, the main addictive component of tobacco, is known to exert physiological effects by binding to its receptor, the nicotinic acetylcholine receptor (nAChR). Recent studies have identified the presence of nAChRs in non-neuronal cells, and in macrophages, functional alteration upon stimulation with nicotine has been documented. To understand the impact of in utero nicotine exposure on various immune cell disposition and function, we designed preliminary studies using an in vivo model of P. aeruginosa infection. In this model, pregnant mice were exposed to nicotine and after weaning, offspring were infected intra-tracheally and humanely killed 5 days later. Nicotine-exposed mice had a greater weight reduction post-infection. This was accompanied by a decreased number of neutrophil, resident macrophages, and B lymphocytes in the lungs, while the number of B lymphocytes in the lymph nodes were greater than that of the control group. In the lung lavage fluids, IL-6, MCP-1, and TNFα concentrations were elevated in nicotine-exposed mice. In an in vitro system using bone marrow-derived macrophages, a significantly reduced production of IFNγ was observed in nicotine-exposed mice when cells were stimulated with LPS. To characterize and compare gene expression in macrophages isolated from neonates developmentally exposed to nicotine, we designed a clinical study to recruit pregnant mothers who 1) did not smoke during pregnancy, 2) smoked throughout pregnancy, or 3) used NRT during pregnancy. We found that successful RNA isolation can be achieved from neonatal tracheal aspirate samples and cell number and reagent volumes were important determinants of acceptable RNA quality and quantity. Together, these preliminary findings demonstrate a possible alteration in immune response as a result of in utero nicotine exposure and sets a groundwork for future studies in identifying mechanisms underlying the impact of developmental nicotine exposure.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The exposure to the cigarette smoke xenobiotics is related with a compromised immune system and the development of many diseases such as Chronic Obstructive Pulmonary Disease (COPD) and lung cancer. The NTPDase and acetylcholinesterase (AChE) are important enzymes involved in the regulation of immune system and alterations in their activities have been showed in many diseases including the related with cigarette smoke-exposure. The curcumin (Cur), polyphenol isolated from the rhizomes of Curcuma longa and widely employed in the culinary and oriental traditional medicine, has many biological activities such as antioxidant, anticarcinogenic and antiinflammatory. Considering the relevance of lymphocytes in the establishment and progression of immune responses, the purpose of this study was to evaluate the effect of curcumin in the NTPDase and AChE activities in peripheral lymphocytes (PL) and lung lymphocytes (LL) from rats exposed to cigarette smoke. The establishment of a protocol to lung lymphocytes separation using the lung tissue was performed with the objective of optimize the cellular viability and the purity of separation, using as references the protocols for separation of human peripheral and lung lymphocytes. The experimental procedure to evaluate the effect of curcumin on the NTPDase and AChE in lymphocytes from rats exposed to the cigarette smoke was divided in two sets of experiments. In the first set, the animals were randomly divided into four groups: Vehicle; Cur 12.5 mg/kg; Cur 25 mg/kg; Cur 50 mg/kg. In the second experimental set, the animals were divided into 5 groups: Vehicle; smoke exposed; smoke and Cur 12.5 mg/kg; smoke and Cur 25 mg/kg; smoke and Cur 50 mg/kg. Curcumin was diluted with corn oil, administered by oral gavage, not exceeding 1 ml/kg body weight. The treatment with curcumin and cigarette smoke was carried out once a day, 5 days each week, during 30 days. It was administered the curcumin or corn oil, and approximately 10 minutes later, the smoking groups were exposed to the sidestream smoke of four commercial cigarettes (nicotine 0.9 mg, tar 10 mg each) inside a whole-body smoke exposure chamber. Control animals were placed in an equal chamber for the same amount of time. After thirty days, the animals were euthanized, the lung was removed and the peripheral blood collected for separation of lymphocytes. With the standardization of the protocol to the separation of lung lymphocytes it was achieved a percentage of lymphocytes higher than 82% and it was observed a strong correlation between the enzyme activity using ATP and ADP as substrates, which indicates that the results can be expressed in milligram of protein, millions of cells and millions of viable cells. The results from the experiments with the cigarette smoke exposure demonstrated that the treatment with curcumin prevented the alterations observed in the cigarette smoke-exposed animals such as the decrease of ATP and ADP hydrolysis in PL and LL, and the increase of AChE activity in PL. We suggest that the treatment with curcumin was protective, since the high concentrations of ATP are positively related with inflammation and tissue damage, and the emphysematous damages observed in cigarette smoke-exposed was not observed in the groups treated with curcumin. / A exposição aos xenobióticos da fumaça do cigarro está relacionada com o comprometimento do sistema imune e o desenvolvimento de diversas doenças como a Doença Pulmonar Obstrutiva Crônica (DPOC) e câncer de pulmão. A NTPDase e a acetilcolinesterase (AChE) são importantes enzimas envolvidas na regulação do sistema imune e alterações nas suas atividades tem sido descritas em várias doenças incluindo as relacionadas à exposição à fumaça do cigarro. A curcumina (Cur), polifenol obtido a partir de rizomas de Curcuma longa e amplamente utilizado na culinária e na medicina tradicional oriental, possui muitas atividades biológicas como antioxidante, anticarcinogênica e antiinflamatória. Tendo em vista a relevância dos linfócitos no estabelecimento e progressão de respostas imunes, o objetivo deste estudo foi avaliar o efeito da curcumina na atividade das enzimas NTPDase e AChE em linfócitos periféricos (PL) e linfócitos pulmonares (LL) de ratos expostos à fumaça do cigarro. A padronização da separação dos linfócitos a partir de tecido pulmonar foi realizada com o objetivo de otimizar a viabilidade celular e a pureza da separação, utilizando-se como base os protocolos de separação de linfócitos periféricos e pulmonares de seres humanos. Os procedimentos experimentais para a avaliação do efeito da curcumina sob a atividade das enzimas NTPDase e AChE em linfócitos de ratos expostos à fumaça do cigarro foram divididos em duas etapas. Primeiramente, os animais foram divididos aleatoriamente em quatro grupos: veículo; Cur 12.5 mg/kg, Cur 25 mg/kg; Cur 50 mg/kg. Na segunda etapa experimental os animais foram divididos aleatoriamente em cinco grupos: veículo; cigarro; cigarro e Cur 12.5 mg/kg; cigarro e Cur 25 mg/kg; cigarro e Cur 50 mg/kg. A curcumina foi diluída em óleo de milho, administrada por gavagem oral, não excedendo 1 ml/kg de massa corpórea. O tratamento com a curcumina e com a fumaça do cigarro foi realizada uma vez por dia, cinco dias por semana, durante trinta dias. A curcumina foi administrada e, após aproximadamente dez minutos, os grupos fumantes eram expostos à fumaça de quatro cigarros comerciais (0,9 mg nicotina, 10 mg alcatrão cada) dentro de uma câmara de exposição. Os animais não expostos à fumaça do cigarro foram colocados em uma câmara equivalente, pelo mesmo período. Após trinta dias, os animais foram eutanasiados, os pulmões removidos e o sangue periférico coletado para a separação dos linfócitos. Com a padronização da técnica para a separação de linfócitos pulmonares foi obtido um percentual de linfócitos superior a 82% e ainda observou-se uma forte correlação entre a atividade da enzima usando ATP e ADP como substrato o que indica que os resultados podem ser expressos tanto em miligramas de proteína, quanto em milhões de células quanto em milhões de células viáveis. Os resultados dos experimentos com a fumaça do cigarro demonstraram que o tratamento com a curcumina preveniu as alterações observadas nos animais expostos à fumaça do cigarro como a diminuição na hidrólise de ATP e ADP em PL e LL, e o aumento da atividade da AChE em PL. Analisando os resultados obtidos, sugere-se que o tratamento com a curcumina foi protetor, uma vez que as altas concentrações de ATP estão positivamente relacionadas com inflamação e dano tecidual, e as lesões enfisematosas observadas no grupo exposto à fumaça do cigarro não foram observadas com os tratamentos com a curcumina.

Le tabagisme actif par la mère expose le fœtus en développement à des agents qui peuvent traverser la barrière placentaire et interférer avec les fonctions placentaires. Un large éventail de fonctions immunologiques, pourrait être compromises. Dans cette étude, nous avons évalué l'effet de l'extrait de la fumée de cigarette (CSE) sur les macrophages isolés à partir de placentas humains (pMφs), qui sont les principaux partenaires de l'immunité de fœto-maternelle innée. J’ai pu montrer que le CSE inhibe la formation des cellules géantes multinucléées (MGC). Cette propriété du CSE est spécifique aux macrophages car la fusion des macrophages dérivés des monocytes est inhibée lors de la formation de granulomes in vitro. J’ai également étudié l'absorption de particules et la production de cytokines par pMφs exposés au CSE. Le CSE a inhibé l'absorption des particules de zymosan, mais pas celle du zymosan opsonisé, ce qui suggère qu’il interfère avec les récepteurs phagocytaires et non phagocytaires. Le CSE augmente la libération de TNF et d'IL-33, et une diminue celle de l'IL-10, ce qui montre que l'équilibre entre les cytokines est affecté par le CSE. En outre, l’expression des métalloprotéinases telles que les MMP-1, MMP-10 et MMP-12, connues pour être impliquées dans le remodelage des tissus et la fusion des macrophages est dérégulée. Enfin, j’ai montré que la nicotine, l'un des principaux composés de tabac, n'a pas affecté les propriétés fonctionnelles des pMφs. / Active smoking by the mother exposes the developing fetus to agents that can cross the placental barrier and interfere with placental functions. A wide range of immunological functions, including innate and adaptive immune responses, might be impaired. In this study, we assessed the effect of cigarette smoke extract (CSE) on macrophages isolated from human placentas (pMφs), which are major partners of innate feto-maternal immunity. I showed that CSE significantly inhibited the formation of multinucleated giant cells (MGCs). This property of CSE is specific to macrophages because the fusion of monocyte-derived macrophages is inhibited during the in vitro formation of granulomas. I also investigated particle uptake and cytokine production by pMφs exposed to CSE. CSE inhibited the uptake of zymosan, but not that of opsonized zymosan, suggesting that it interferes with phagocytic receptors, not with the phagocytic machinery of pMφs. CSE increased the release of Tumor Necrosis Factor and interleukin-33, and decreased that of interleukin-10, demonstrating that the balance between inflammatory and anti-inflammatory cytokines is affected by CSE. Furthermore, CSE enhanced the expression of metalloproteinase (MMPs) genes such as MMP-1, MMP-10 and MMP-12, known to be involved in tissue remodeling including macrophage fusion. Finally, I showed that nicotine, one of the major compounds of tobacco, did not affect the functional properties of pMφs.