Escala de Resistencia a la Presión de Pares para el Consumo de Cigarro Convencional en estudiantes universitarios: adaptación y validación / Resistance to Peer Pressure for Cigarette Smoking in University Students: Scale Adaptation and Validation / Escala de resistência à pressão dos pares para o consumo de cigarros em universitários: adaptação e validação

Introducción: la influencia o presión de pares que fuman es uno de los principales factores por los que los estudiantes universitarios inician el consumo del cigarro convencional; sin embargo, no se ha encontrado un instrumento que evalúe este fenómeno. Por lo tanto, el objetivo fue adaptar y validar la Escala de Resistencia a la Presión de Pares para el Consumo de Cigarro Convencional. Materiales y métodos: participaron 237 estudiantes universitarios del estado de Nuevo León (México), de 18 a 24 años. Se realizó análisis factorial exploratorio, análisis de confiabilidad, correlación de Spearman y prueba de Kruskal-Wallis. Resultados: el 63.3 % de los estudiantes fueron mujeres y la media de edad fue de 19.66 años. Se identificaron dos factores con un total de 13 ítems. Se obtuvo un alfa de Cronbach de 0.81. Se encontraron diferencias estadísticamente significativas entre los distintos tipos de consumidores de cigarro convencional y los puntajes de la escala de resistencia a la presión de pares (H[4] = 23.85; p < 0.001). Conclusiones: la nueva versión de la Escala de Resistencia a la Presión de Pares para el Consumo de Cigarro Convencional evidenció adecuadas propiedades psicométricas para evaluar la presión que ejercen los pares en estudiantes universitarios para el consumo de cigarro convencional

Introduction: Influence or peer pressure is one of the leading factors in developing cigarette smoking habits in university students; however, no effective strategy to assess this phenomenon has been developed yet. This study aimed to adapt and validate the peer pressure resistance scale to conventional cigarette consumption. Materials and methods: A total of 237 university students from the Nuevo León State (Mexico), aged 18­24 years, were enrolled. Exploratory factor and reliability analyses, the Spearman correlation, and the Kruskal­Wallis test were performed. Results: 63.3% of the students were women, and the mean age was 19.66 years. The exploratory analysis extracted two factors with a total of 13 items. A Cronbach's Alpha of 0.81 was found. Statistically significant differences were found between the different types of conventional cigarette users and peer pressure resistance scale scores [H(4) = 23.85; p < .001] were found. Conclusions: The peer pressure resistance scale showed appropriate psychometric properties for assessing the peer pressure to smoke conventional cigarettes in university students.

Introdução: a influência ou pressão dos pares que fumam é um dos principais fatores que levam os universitários a começarem a fumar cigarros convencionais, porém não foi encontrado nenhum instrumento para avaliar esse fenômeno. Portanto, o objetivo do trabalho foi adaptar e validar a escala de resistência à pressão dos pares para o consumo de cigarro convencional. Materiais e métodos: participaram 237 estudantes universitários do estado de Nuevo León, México, de 18 a 24 anos. Foram realizadas análise fatorial exploratória, análise de confiabilidade, correlação de Spearman e teste de Kruskal-Wallis. Resultados: 63,3% dos alunos eram mulheres e a média de idade foi de 19,66 anos. Dois fatores foram identificados com um total de 13 itens. Obteve-se um alfa de Cronbach de 0,81. Diferenças estatisticamente significativas foram encontradas entre os diferentes tipos de usuários de cigarros convencionais e as pontuações na escala de resistência à pressão dos pares (H(4) = 23,85; p < 0,001). Conclusões: a nova versão da escala de resistência à pressão dos pares para o consumo de cigarros convencionais apresentou propriedades psicométricas adequadas para avaliar a pressão exercida pelos pares sobre os universitários para o consumo de cigarros convencionais

Effect of Euphorbia helioscopia Aqueous Extract on Chronic Obstructive Pulmonary Disease and Precancerous Lesion-associated Proteins of Lung Tissues in Mice Induced by Cigarette Smoke / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect and mechanism of Euphorbia helioscopia aqueous extract （EHE） on mice with chronic obstructive pulmonary disease （COPD） and its influence on precancerous lesion-associated proteins in lung tissues induced by cigarette smoke （CS）. MethodThe COPD model was induced by CS in 60 mice and the model mice were randomly divided into control group， model group， positive drug group （dexamethasone， 2 mg·kg-1）， and low-， medium-， and high-dose EHE groups （1.875， 3.75， 7.5 g·kg-1）. The high-performance liquid chromatography （HPLC） method was used to determine the related components in EHE. The changes in end-expiratory pause （EEP）， airway resistance （Penh）， expiratory flow at 50% vital capacity （EF50）， and other pulmonary function indexes were detected by the spirometer. The levels of inflammatory factors， such as interleukin （IL）-2， IL-5， IL-18， IL-17A， and IL-27 in bronchoalveolar lavage fluid （BALF） of mice were detected by high-throughput liquid protein chip technology. Hematoxylin-eosin （HE） staining was used to detect the pathological changes in lung tissues in mice. The content of malondialdehyde （MDA）， myeloperoxidase （MPO）， and glutathione peroxidase （GSH-Px） in lung tissues was determined by the colorimetric method. The mRNA relative expression of tumor necrosis factor-α （TNF-α）， transforming growth factor-β （TGF-β）， matrix metalloproteinase-2 （MMP-2）， matrix metalloproteinase-9 （MMP-9）， and matrix metalloproteinase-12 （MMP-12） was detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Immunohistochemistry （IHC） was used to detect the expression of tumor protein （P53） and cell proliferation-associated antigen （Ki67） in lung tissues， and Western blot was used to detect the relative expression of tumor suppressor protein （P16）， DNA （cytosine-5）-methyltransferase 1 （DNMT1）， and fragile histidine triad （FHIT） in lung tissues. ResultThe results showed that the main compounds in EHE included phenols （gallic acid and protocatechuic acid） and flavonoids （such as hyperoside， rutin， myricetin， naringenin， quercetin， luteolin， kaempferol， and licorice chalcone A）， among which gallic acid and rutin were the highest in content. Compared with normal group， model group showed increased levels of EEP， EF50， and Penh （P<0.05）， and showed increased MDA and MPO levels （P<0.01） and decreased GSH-Px （P<0.01）， and the model group displayed increased levels of IL-2， IL-5， IL-18， IL-17A， IL-27， TNF-α， TGF-β， MMP-2， MMP-9， and MMP-12 （P<0.05）. And the model group exhibited up-regulated expression of P53， Ki67， and FHIT in lung tissues （P<0.01） and down-regulated expression of DNMT1 and P16 （P<0.01）. Compared with model group， the EHE groups showed decreased EEP and EF50 levels （P<0.05）. The pathological injury of lung tissues in mice of the model group was observed under HE staining， and the pathological injury of basal cell hyperplasia of lung tissues was gradually improved after treatment with EHE. The EHE groups showed reduced levels of MDA and MPO （P<0.01） and increased GSH-Px （P<0.01）. The EHE groups displayed decreased levels of IL-2， IL-5， IL-18， IL-17A， IL-27， TNF-α， TGF-β， MMP-2， MMP-9， and MMP-12 （P<0.05）. And the EHE groups showed down-regulated Ki67 and FHIT in lung tissues （P<0.05） and up-regulated expression of P53 and DNMT1 （P<0.05）. ConclusionEHE can protect mice from COPD and inhibit precancerous lesions， and the mechanism may be related to the inhibition of inflammation and oxidative stress response， regulation of protease and antiprotease imbalance， and regulation of epithelial cell growth.

Cigarette Smoke Induces Gefitinib Resistance in NSCLC Cells via ROS/Sirt3/SOD2 Pathway / 中国肺癌杂志

BACKGROUND@#Epidermal growth factor receptor (EGFR) gene mutations are the most common driver mutations in non-small cell lung cancer (NSCLC). To prolong the survival of the patients, EGFR tyrosine kinase inhibitors (TKIs) resistance in NSCLC is a major challenge that needs to be addressed urgently, and this study focuses on investigating the mechanism of cigarette smoke (CS) induced Gefitinib resistance in NSCLC.@*METHODS@#PC-9 and A549 cells were cultured in vitro and treated with 1 µmol/L Gefitinib for 4 h and 10% cigarette smoke extract (CSE) for 48 h. Western blot was used to detect Sirtuin 3 (Sirt3) and superoxide dismutase 2 (SOD2) protein expressions; DCFH-DA probe was used to detect intracellular reactive oxygen species (ROS); CCK-8 kit was used to detect cell activity, and EdU was used to detect cell proliferation ability. Sirt3 overexpression plasmid (OV-Sirt3) was transfected in PC-9 and A549 cells and treated with 1 µmol/L Gefitinib for 4 h and 10% CSE for 48 h after N-acetylcysteine (NAC) action. The expressions of Sirt3 and SOD2 were detected by Western blot; the ROS level in the cells was detected by DCFH-DA probe, and the cell activity was detected by CCK-8.@*RESULTS@#CSE induced an increase in the 50% inhibitory concentration (IC50) of both PC-9 and A549 cells to Gefitinib (P<0.01) and enhanced the proliferation of PC-9 and A549 cells, suggesting that CS induced Gefitinib resistance in NSCLC. ROS was involved in CSE-induced Gefitinib resistance (P<0.05). CSE induced low expressions of Sirt3 and SOD2 (P<0.01), and Sirt3/SOD2 was associated with poor prognosis in lung cancer patients (P<0.05). OV-Sirt3 in PC-9 and A549 cells reversed CSE-induced Gefitinib resistance (P<0.05) and significantly reduced ROS production. NAC reversed CSE-induced Gefitinib resistance in PC-9 and A549 cells (P<0.05).@*CONCLUSIONS@#The ROS/Sirt3/SOD2 pathway is involved in CS-induced Gefitinib resistance in NSCLC.

Effect of zinc and probiotics supplementation on IL-6 and tissue neutrophil levels in rats exposed to cigarette smoke

@#Introduction: Cigarette smoke exposure can cause inflammation, inducing the release of acute phase cytokines, such as IL-6, that will then trigger the recruitment of neutrophils, which are mostly phagocytic cells. Zinc and probiotics are known to have beneficial effects against inflammation. This study was conducted to investigate the effect of zinc and probiotics supplementation on IL-6 and tissue neutrophil levels in rats exposed to cigarette smoke. Methods: In a randomised, experimental study with post-test control group design, thirty 2 to 3-month-old male Wistar rats, each weighing 180-220 g, were divided into five groups: control group without treatment (C); exposed to cigarette smoke [C (-)]; exposed to cigarette smoke and received zinc (Z); exposed to cigarette smoke and received probiotics (P); and exposed to cigarette smoke and received a combination of zinc and probiotics (ZP). Results: Mean tissue neutrophil levels in Z, P, and ZP groups were 43.43±2.01, 34.67±1.32,and 29.77±5.05 cells, respectively. There were significant differences between supplementation intake and tissue neutrophil levels in each group compared to C (-) group (p<0.05). Meanwhile, only IL-6 level in the ZP group (6.02 pg/mL) decreased significantly compared to C (-) group (10.61 pg/mL). Conclusion: These results suggest that a combination of zinc and probiotics have an anti-inflammatory effect as measured by IL-6 and neutrophil levels.

Dendrobium Polysaccharides Mitigate CSE-induced Inflammatory Damage of Human Bronchial Epithelial Cells via TLR4/NF-κB Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effects of Dendrobium polysaccharides on the secretion of inflammatory cytokines and Toll-like receptor 4 （TLR4）/nuclear factor （NF）-κB pathway in 16HBE cells exposed to cigarette smoke extract （CSE）. MethodThe 16HBE cells were classified into the control， CSE， and CSE+ Dendrobium polysaccharides （100， 200， 400 mg·L-1） groups. The cell-counting kit-8 （CCK-8） assay was employed to measure the cell viability， and a microscope was used to observe the cell morphology. The enzyme-linked immunosorbent assay was employed to measure the levels of interleukin （IL）-8， IL-1β， IL-4， IL-13， and transforming growth factor （TGF）-β in cell culture supernatants. Real-time PCR was carried out to determine the mRNA levels of Toll-like receptor 4 （TLR4）， nuclear factor-κB （NF-κB）， and IL-4. Western blot was employed to determine the protein levels of interleukin-4 receptor （IL-4R）， TLR4， myeloid differentiation primary response protein 88 （MyD88）， NF-κB， phosphorylated nuclear factor-κB （p-NF-κB）， and nucleoproteins nuclear factor-κB （NEs-NF-κB）. The immunofluorescence assay was employed to measure the nuclear translocation of NF-κB. ResultCompared with the control group， the CSE group showed elevated levels of IL-8， IL-1β， IL-4， IL-13， and TGF-β in the cell culture supernatants （P<0.05， P<0.01）， up-regulated expression levels of TLR4， MyD88， NF-κB， p-NF-κB， NEs-NF-κB， and IL-4 （P<0.01）， and significant nuclear translocation of NF-κB. Compared with the CSE group， Dendrobium polysaccharides increased the cell survival rate， recovered the cell activity， lowered the levels of IL-8， IL-1β， IL-4， IL-13， and TGF-β， down-regulated the expression of TLR4， MyD88， NF-κB， p-NF-κB， NEs-NF-κB， and IL-4 （P<0.05， P<0.01）， and reduced the nuclear translocation of NF-κB. ConclusionDendrobium polysaccharides showed significant protective effects on the 16HBE cells exposed to CSE by inhibiting the TLR4/NF-κB signaling pathway.

Impact of cigarette smoke on osteogenic and osteoclast signaling in middle palatal suture

Abstract Considering that smoking is a public health problem that has been growing among adolescents, the aim of this study was to investigate the impact of cigarette smoke on osteogenic and osteoclastogenic signaling in middle palatal suture of rats. Male Wistar rats exposed (n = 30) or not to cigarette smoke (n = 30) were used. Exposure to smoke was carried out for two daily periods of 3 minutes each, with an interval of 12 hours between exposures. After the experimental periods of 3, 7, 14 and 21 days, the animals were euthanized. The collected tissues were analyzed using light microscopy and real-time RT-PCR was performed to investigate gene expression. The data obtained were compared using the Kruskal Wallis and Dunn tests (⍺ = 5%). Morphologically, there were no significant changes in the middle palatal suture of rats exposed or not to cigarette smoke during 3, 7, 14 and 21 days (p> 0.05). On the other hand, osteoclastogenic signaling was increased in animals exposed to smoke and was characterized by a higher production of RANKL at 3 and 14 days (p <0.05), with no change in the synthesis of RANK and osteoprotegerin (p> 0.05). Interestingly, in the exposed animals, an early increase in the synthesis of osteocalcin, bone sialoprotein and osteopontin was also identified at 3 days of exposure (p <0.05), not sustained over time (p> 0.05). Cigarette smoke modulates osteogenic and osteoclastogenic signaling in the middle palatal suture of young rats, although morphological changes have not been evidenced.

Resumo Considerando que a fumaça de cigarro é um problema de saúde pública que está crescendo entre os adolescentes, o objetivo deste estudo foi investigar o impacto da fumaça de cigarro na sinalização osteogênica e osteoclastogênica da sutura palatina mediana de ratos. Foram utilizados ratos Wistar machos expostos (n=30) e não expostos à fumaça de cigarro (n=30). A exposição à fumaça de cigarro foi realizada duas vezes ao dia por 3 minutos, com um intervalo de 12 horas entre as exposições. Os animais foram mortos após o período experimental de 3, 7, 14 e 21 dias. Os tecidos coletados foram analisados em microscópico de luz e pelo RT-PCR em tempo real foi realizado para investigar a expressão gênica. s dados obtidos foram comparados usando o testes de Kruskal Wallis e Dunn (⍺ = 5%). Morfoligicamente, não houve mudança significativa na sutura palatina mediana nos ratos expostos ou não à fumaça de cigarro durante os tempo de 3, 7, 14 e 21 dias (p> 0.05). Por outro lado, a sinalização osteogênica esta aumentada nos animais expostos à fumaça e foi caracterizado por um aumento da produção de RANKL aos 3 e 14 dias (p <0.05), sem mudança na síntese da produção de RANK e osteoprotegerina (p> 0.05). Curiosamente, nos animais expostos, também foi observado um aumento precoce da síntese de osteocalcina, sialoproteína óssea e de osteopontina aos 3 dias de exposição, o que não foi mantido ao longo do tempo. A fumaça de cigarro modula a sinalização osteogênica e osteoclastogênica na sutura palatina mediana de ratos jovens, apesar de não tenha sido evidenciado alterações morfológicas.

Bufei Yishen formula attenuates cigarette smoke extract-induced airway mucus hypersecretion by regulating Notch signaling pathway / 中华危重病急救医学

Objective:To explore the mechanism of Bufei Yishen formula (BYF) on attenuating cigarette smoke extract (CSE)-induced airway mucus hypersecretion by regulating Notch signaling pathway.Methods:The human airway epithelial cell 16HBE was cultured in vitro, and the cells in logarithmic growth phase were used for the experiments. ① Intervention condition screening experiment: the 16HBE cells were grouped, methylthiazolyldiphenyl-tetrazolium (MTT) method and enzyme-linked immunosorbent assay (ELISA) were used to detect the effects of different concentrations of CSE (2.5%, 5%, 10%, 20%, 40%), different concentrations of BYF drug-containing serum (5%, 10%, 20%, 40%), and different concentrations of Notch signal pathway blocker DAPT (5, 10, 20, 40 μmol/L) on cell activity and secretion of mucin 5AC (MUC5AC) levels. In addition, a blank control group was set up to screen out the best conditions for preparing CSE-induced cell mucus hypersecretion model and BYF and DAPT intervention. ② Intervention experiment: the 16HBE cells were divided into four groups. The blank control group was not given any treatment; the 16HBE cells were induced by 10% CSE for 24 hours to prepare mucus hypersecretion model in the CSE model group; the cells in the CSE+BYF group and CSE+DAPT group were given 10% BYF or 20 μmol/L DAPT, respectively, for intervention at the same time for 24 hours. Real-time fluorescent quantitative polymerase chain reaction (qPCR) was used to detect the mRNA expressions of MUC5AC, Notch3 and hairy and enhancer of split 1 (HES1) in the cells. Western blotting was used to detect the protein expressions of Notch3 and HES1 in the cells. Results:① Results of the screening experiment of intervention conditions: compared with the blank control group, 10% CSE induction for 24 hours was the best condition for establishing cell mucus hypersecretion model that neither affected cell viability nor increased the secretion of MUC5AC; while 10% BYF and 20 μmol/L DAPT was the optimal intervention condition. ② Intervention experiment results: compared with the blank control group, the mRNA expressions of MUC5AC, Notch3, and HES1 and the protein expressions of Notch3 and HES1 in the CSE model group were significantly increased, indicating that CSE activated Notch3 and HES1 signal activation and induced 16HBE cells to secrete mucus protein. Compared with the CSE model group, BYF and DAPT could significantly down-regulate the mRNA and protein expressions of MUC5AC, Notch3, and HES1 in cells [MUC5AC mRNA (2 -ΔΔCT): 1.03±0.13, 0.96±0.05 vs. 1.35±0.07, Notch3 mRNA (2 -ΔΔCT): 1.10±0.14, 1.10±0.02 vs. 1.31±0.15, HES1 mRNA (2 -ΔΔCT): 1.26±0.10, 1.14±0.15 vs. 1.45±0.08, Notch3 protein (Notch3/GAPDH): 0.10±0.03, 0.16±0.03 vs. 0.31±0.09, HES1 protein (HES1/GAPDH): 0.37±0.06, 0.34±0.08 vs. 0.50±0.05, all P < 0.05]. Conclusion:The mechanism of BYF attenuating mucus hypersecretion of 16HBE cells induced by CSE was associated with the inhibition of Notch signaling pathway activation.

Progress on associations between intrauterine cigarette smoke exposure and cognitive and behavioral development / 国际儿科学杂志

Intrauterine cigarette smoke exposure(ICSE)refers to a condition under which pregnant women actively inhale and/or passively inhale cigarettes and smogs mostly consisting of nicotine, tar and carbon monoxide during the first, second and third trimesters of pregnancies.As an adverse environmental factor in early life, ICSE is associated with increased risks of various diseases at childhood and adulthood.ICSE is closely related with children′s cognitive and behavioral developmental disorders.It has been reported that ICSE led to elevated risks of cognitive impairments including disorders in fine motor skills, language and vocabulary, reading comprehension, matrix reasoning, working memory, learning skills, orientation and other capabilities.Additionally, it is associated with increased risks of attention deficit hyperactivity disorder and sleep disorder, but the association with autism spectrum disorder is still controversial.The review focuses on the effects of ICSE on cognitive and behavioral development, and summarizes the underlying mechanisms, providing more clinical ideas for etiological studies of cognitive and behavioral developmental disorders in children.

Chronic exposure to cigarette smoke transiently worsens the disease course in a mouse model of pulmonary paracoccidioidomycosis

ABSTRACT Paracoccidioidomycosis (PCM) may present as an acute/subacute clinical form, characterized by a progressive disease arising from the airborne initial infection, or, most often, as an asymptomatic or subclinical infection that may manifest later during an individual's life, the chronic form. Epidemiological studies show the existence of a strong association between smoking and the development of the chronic form. Current evidence demonstrates that cigarette smoke (CS) has immunosuppressive properties that could be implicated in the increasing susceptibility to the chronic form of PCM. To address this issue, we developed a murine model of a non-progressive pulmonary form of PCM that was exposed to CS at a magnitude that mimicked a moderate smoker. The chronic CS exposure started after 2 weeks and lasted up until 20 weeks post-infection, with the aim of mimicking human natural history, since it is estimated that individuals from endemic areas are infected early in life. The control group consisted of infected but not CS-exposed mice. We assessed the lung fungal burden (colony forming units [CFU]) and the area affected by the granulomatous inflammatory response, fungal dissemination to spleen and liver, and, by immunohistochemistry, the presence of CD4 and CD8 lymphocytes, CD68 and MAC-2 macrophages, and IFN-γ, IL-10 and TNF expressing cells within the granulomatous response. We detected a CS effect as early as 2 weeks after exposure (four weeks post-infection) when the lung CFU of exposed animals was significantly higher than in their non-exposed counterparts. At 12 weeks, the CS-exposed animals presented a more severe disease, as witnessed by the persistent higher lung fungal load (although it did not reach statistical significance [ p = 0.054]), greater dissemination to other organs, greater affected area of the lung, decreased IFN-γ/IL-10 ratio, and higher TNF expression within the granulomas, compared with CS-non-exposed mice. The number of CD4 and CD8 lymphocytes infiltrating the granulomas was similar between both mice groups, but there was a decrease in the number of MAC-2+ macrophages. No difference was noted in the CD68+ macrophage number. However, the follow-up in week 20 showed that the immunological effects of exposure to CS ceased, with both CS and NCS mice showing the same infectious features, i.e., a trend for resolution of the infection. In conclusion, we show that chronic CS-exposure alters the course of the disease in an experimental model of subclinical pulmonary PCM, confirming the epidemiological link between CS-exposure and the chronic form of PCM. However, we also show that this effect is transitory, being detected between 4- and 12-weeks post-infection but not thereafter. The possible immune mechanisms that mediate this effect and the reasons for its transitory effect are discussed.

Differentially expressed microRNAs in the testis of male rats exposed to cigarette smoke: Screening and function verification / 中华男科学杂志

Objective@#To screen differentially expressed miRNAs in the testis of male rats exposed to cigarette smoke (CS) and identify the early molecular markers of CS-induced apoptosis of testicular cells.@*METHODS@#We randomly divided 200 SPF male SD rats into blank control and low-dose (10 non-filter cigarettes/d), medium-dose (20 non-filter cigarettes/d) and high-dose (30 non-filter cigarettes/d) CS exposure groups. After 2, 4, 6, 8 and 12 weeks of CS exposure, we observed the histopathological changes of the testis by HE staining, detected the apoptosis of the testicular cells by TUNEL, and determined the expressions of caspase-3 and caspase-9 in the testis tissue by immunohistochemistry, RT-PCR and Western blot. Based on the laboratory results, we selected 4 testicular samples from the 12-week high-dose group and another 4 from the control for miRNA microarray-based screening, bioinformatics analysis, and verification of differentially expressed miRNAs in all the animals by RT-PCR.@*RESULTS@#Compared with the controls, the CS-exposed rats showed dose- and time-dependent increase in the atrophy of the testis and significantly increased number of apoptotic testis cells from the 6th week of exposure (P < 0.05), with dramatically up-regulated expressions of caspase-3 (P < 0.01) and caspase-9 protein and mRNA (P < 0.05) in the testis tissue. Microarray-based screening and RT-PCR revealed 5 differentially expressed miRNAs in the testis of the CS-exposed rats, of which miR-138-5p, miR-181d-5p, miR-19a-3p and miR-3588 were down-regulated, and miR-155-5p up-regulated, and the target genes of the differentially expressed miRNAs positively regulated the apoptosis of the testicular cells.@*CONCLUSIONS@#The differentially expressed miRNAs miR-155-5p, miR-138-5p, miR-181d-5p, miR-19a-3p and miR-3588 regulate CS-induced apoptosis of testicular cells, and may become biomarkers for early diagnosis and prognosis of CS-induced spermatogenesis obstruction.《.

Effect of erosive challenge and cigarette smoke on dentin microhardness, surface morphology and bond strength / Efeito de desafios erosivos e fumaça de cigarro na microdureza, morfologia de superfície e resistência de união à dentina

Objective: This study aimed to evaluate the surface microhardness and morphology, as well as the microshear bond strength of a self-etching adhesive (Clearfil SE, Kuraray) to eroded dentin, exposed or not to cigarette smoke. Forty dental crowns were divided into 4 groups (n = 10): no treatment (control) (C); erosion (E); erosion + cigarette smoke exposure (ES); cigarette smoke exposure (S). Samples were prepared through third molars polishing until dentin exposure, followed by crown section. Erosive cycles were performed 5 times/day for 30 s at 60 min intervals. Cigarette smoke was produced with twenty cigarettes/day, during 5 days. Microhardness was evaluated initially and after the treatments. Microshear bond strength was tested after the treatments and dentin restoration with flow composite. Failure patterns and dentin morphology was evaluated by Scanning Electron Microscopy. Microshear bond strength data was submitted to two-way ANOVA, microharness test was adjusted by gamma distribution to be a non-parametric analyses (p=0.05), and surface morphology as qualitative analyses. Loss percentage of microhardness was observed only in groups submitted to erosion. Bond strength was statistically similar between all groups. The most prevalent failure pattern was of adhesive type. Morphological analysis of dentin showed obliterated tubules in groups submitted to cigarette smoke exposure. Cigarette smoke exposure did not promote any effect in the percentage of microhardness loss, as in sound dentin as in eroded dentin. Cigarette smoke, erosion, and association of both, did not alter the bond strength of self-etching adhesives to dentin. (AU)

Objetivo: Este estudo teve como objetivo avaliar a microdureza (% perda de dureza) e morfologia de superfície (MS), assim como a resistência de união (RU) de um adesivo autocondicionante (Clearfil SE, Kuraray) à uma dentina erodida, exposta ou não à fumaça de cigarro. Material e Métodos: Quarenta coroas dentais de terceiros molares foram seccionadas da raiz e polidas até a exposição dentinária, sendo aleatoriamente divididas em 4 grupos (n=10): sem tratamento (controle), erosão (E), erosão+ exposição a fumaça de cigarro (ES); exposição a fumaça de cigarro (S).O ciclo erosivo foi realizado 5 vezes/dia por 30s, com 60 minutos de intervalo entre eles. Os grupos ES e S foram exposto à fumaça de cigarro produzida por 20 cigarros/dia, durante 5 dias. A avaliação da microdureza foi realizada antes e após os tratamentos, enquanto a resistência da união por microcisalhamento foi realizada após os tratamentos Os padrões de fratura representativos e a MS dentinária foram avaliados por microscopia eletrônica de varredura (MEV). Os dados de RU foram analisados por ANOVA dois fatores, enquanto a análise de microdureza foi ajustada por distribuição gama por ser uma análise não-paramétrica (p=0.05). A MS foi analisada qualitativamente. Resultados: Os grupos expostos aos ciclos erosivos (E e ES) apresentaram % de perda de dureza significativamente menor que os grupos não expostos (Controle e S. aos ciclos erosivos (E e ES). Para RU, não houve diferença estatística significativa entre os grupos. O padrão de fratura mais observado foi do tipo adesivo, e através das imagens obtidas por MEV, observou-se a obliteração de túbulos dentinários no grupo exposto à fumaça de cigarro, enquanto os grupos submetidos aos ciclos erosivos (E e ES) apresentaram maior exposição e diâmetro de túbulos dentinários. Conclusão: A exposição à fumaça de cigarro não promove nenhum efeito quanto a perda de porcentagem de dureza dentinária, assim como em dentina erodida e saudável. A fumaça de cigarro, o processo erosivo, e a associação de ambos, não altera a resistência da união de adesivos autocondicionantes à dentina. (AU)

Research progress in cell transformation test and its application in cigarette smoke evaluation / 公共卫生与预防医学

In order to understand the current development of cell transformation assay (CTA) and its application in the evaluation of cigarette smoke carcinogenesis, the relevant literatures were analyzed and combed from these two aspects. CTA can evaluate the carcinogenicity of various genotoxic and non-genotoxic carcinogens in a short period of time, and has a strong consistency with the results of animal carcinogenic test. After malignant transformation, the cells show changes in cell morphology, immortalization of cells, disappearance of cell-cell contact inhibition, and the ability to form tumors when injected into animals. The identification methods of transformed cells include transformed cell focus count, agglutination test, soft agar culture and inoculation of nude mice, etc. At present, BALB/c 3T3 cells, Bhas 42 cells and SHE cells are the most widely used cells for CTA. Cigarette smoke is a complex aerosol containing a variety of non-genetic carcinogenic chemicals. Cell transformation tests are often used as an in vitro alternative method to evaluate the carcinogenic effects of cigarette smoke, which is different from the short-term genetic toxicity test. It simulates the long-term state of human smoking induced malignant transformation of cells, through the long-term exposure of cells for several decades, which is closer to the occurrence of cancer caused by human smoking. Therefore, CTA can evaluate the carcinogenicity of cigarette smoke and other tobacco products.

Efeitos da exposição à fumaça do cigarro e suas implicações na neuroinflamação / Effects of exposure to cigarette smoke and its implications on neuroinflammation

O Sistema Nervoso Central (SNC) humano é formado por cerca de 86,1 bilhões de neurônios entre o encéfalo e a medula espinhal. O desenvolvimento pré-natal humano (tempo da concepção ao nascimento) possui cerca de 38 semanas, e é dividido na fase embrionária que corresponde ao período das 8 semanas iniciais da gestação, seguido pela fase fetal. A fase embrionária é o período mais vulnerável à ocorrência de anormalidades congênitas. Por ser um órgão com grande período de desenvolvimento, o SNC está sujeito às alterações genéticas, epigenéticas e ambientais. Durante a fase de implantação do embrião, o DNA é mais vulnerável às influências externas, como à fumaça do cigarro, aumentando o risco de retardo do desenvolvimento fetal, o risco de morte súbita pós-natal e de anormalidades do sistema imune. Neste contexto, o objetivo deste trabalho é avaliar os efeitos da exposição à fumaça do cigarro sobre o processo de neuroinflamação da prole de camundongos C57BL/6 expostos à fumaça do cigarro durante a gestação e desafiados ou não com LPS. Para tanto, camundongos C57BL/6 fêmeas prenhes foram expostas à fumaça do cigarro desde o plug vaginal até o nascimento da prole. No 3º dia de vida, os filhotes foram separados para três linhas de trabalho: 1) in vivo: os animais foram desafiados com LPS pelo período de 4h, seguidos de eutanasia e análises de PCR Array do SNC. 2) in vitro: os encéfalos dissecados foram utilizados para a preparação de cultura mista de glia e da cultura enriquecida com neurônio. Após a maturação celular, as células foram estimuladas com LPS 100 ng/mL e, após 24h, foram realizados ensaios de CBA, citometria de fluxo, PCR, dosagem de NO, avaliação de morte celular e metilação global. 3) Encefalomielite Autoimune Experimental (EAE): após o desmame, os animais foram mantidos em suas caixas moradia por 8 semanas sem nenhum estímulo externo, e então foram imunizados com MOG35-55 para o desenvolvimento da EAE. Nos experimentos in vivo observamos o aumento da transcrição de genes relacionados ao processo inflamatório, como interleucinas e quimiocinas. Em relação aos experimentos in vitro observamos maior crescimento de células astrocitárias (astrogliose), e células da microglia com aumento de moléculas co-estimuladoras (CD80 e CD86) bem como da transcrição e concentração de citocinas pró-inflamatórias e produção de NO. Em cultura enriquecida de neurônio, foi observado aumento na porcentagem de células em apoptose no grupo exposto à fumaça do cigarro desafiados ou não com LPS. O bloqueio da atividade da microglia pela minociclina reverteu a apoptose e diminuiu a produção de NO minimizando a morte celular. Em relação aos experimentos de EAE, os animais expostos à fumaça do cigarro no período gestacional, quando imunizados na vida adulta apresentam aumento no grau da doença bem como maior persistência da mesma quando observado escore clínico, além de acompanhados de um grau maior de infiltrado celular e desmielinização. Desta forma podemos concluir que a exposição à fumaça do cigarro durante o período gestacional leva a uma programação fetal com aumento da resposta neuroinflamatória frente a um estimulo sistêmico, trazendo consequências na vida adulta

The human central nervous system (CNS) is made up of about 86.1 billion neurons between the brain and the spinal cord. The human prenatal development (time from conception to birth) is about 38 weeks, and is divided into the embryonic phase that corresponds to the period of the initial 8 weeks of gestation, followed by the fetal phase. The embryonic stage is the period most vulnerable to the occurrence of congenital abnormalities. Because it is an organ with a long period of development, the CNS is subject to genetic, epigenetic and environmental changes. During the embryo implantation phase, DNA is more vulnerable to external influences such as cigarette smoke, increasing the risk of delay on fetal development, risk of sudden postnatal death, and abnormalities of the immune system. In this context, the aim of this work is to evaluate the effects of exposure to cigarette smoke on the neuroinflammation process of offspring of C57BL/6 mice exposed to cigarette smoke during gestation and challenged or not with LPS. For this, pregnant female C57BL/6 mice were exposed to cigarette smoke from vaginal plug to offspring birth. On the 3rd day of life the offspring were separated into three lines of work: 1) in vivo: the animals were challenged with 1mg/Kg LPS and after 4h they followed to euthanasia; PCR analysis of the CNS was made in this period. 2) in vitro: dissected encephalons were used for the preparation of mixed culture of glia and the culture enriched with neuron. After cell maturation, the cells were stimulated with 100 ng/mL LPS and, after 24 hours, CBA, flow cytometry, PCR, NO assay, cell death and global methylation assays were performed. 3) Experimental Autoimmune Encephalomyelitis (EAE): After weaning, the animals were kept in their housing for 8 weeks without any external stimulus, and then were immunized with MOG35-55 for the development of EAE. In the in vivo experiments we observed increased transcription of genes related to the inflammatory process, such as interleukins and chemokines. In vitro experiments showed higher growth of astrocytes (astrogliosis) and microglia cells with increased stimulatory molecules (CD80 and CD86) as well as the transcription and concentration of proinflammatory cytokines and NO production. In the enriched neuron culture, an increase in the percentage of cells in apoptosis was observed in the group exposed to cigarette smoke challenged or not with LPS. Blocking microglial activity by minocycline reversed apoptosis and decreased NO production by minimizing cell death. The EAE experiments shows that the animals exposed to cigarette smoke in the gestational period, when immunized in adulthood, present an increase in the degree of the disease as well as a greater persistence of the disease; The higher as the clinical score higher is the degree of cellular infiltration and demyelination. In this way we can conclude that the exposure to cigarette smoke during the gestational period leads to a fetal programming with increased neuroinflammatory response to a systemic stimulus and that this is able to last until the adult stage

Mineral composition, histomorphometry, and bone biomechanical properties are improved with probiotic, prebiotic, and symbiotic supplementation in rats chronically exposed to passive smoking: a randomized pre-clinical study / Composição mineral, histomorfometria e propriedades biomecânicas óssea são melhoradas com a suplementação de probióticos, prebióticos e simbióticos em ratos expostos cronicamente ao tabagismo passivo: estudo pré-clínico randomizado

ABSTRACT: Cigarette smoke in large centers is one of the most important causes of chronic inflammatory diseases in public health and is associated with a decrease in bone mass, consolidation process, and bone remodeling. Due to their ability to improve intestinal absorption and compete with pathogenic microorganisms, dietary supplementation with functional foods may contribute to improvement in bone quality. Therefore, the objective of this study was to evaluate the effects of functional, probiotic, prebiotic, or symbiotic food supplementation on mineral composition, histomorphometry, and bone biomechanical properties of rats in the growth phase, chronically exposed to cigarette smoke (T).Sixty-four young male rats were randomly assigned to eight groups (n=8): control (C) [standard diet (SD)]; probiotic (Pro) [SD + probiotic (Lactobacillus acidophilus, Enterococcus faecium, Bifidobacterium thermophilum and Bifidobacterium longum) (2-5 109 UFC each)]; prebiotic (Pre) [SD+ prebiotic (mannan oligosaccharide)]; symbiotic (Sym) (SD + probiotic + prebiotic); control smoking (SC) [(SD + exposure protocol to passive smoking (PS)]; probiotic smoking (ProS) (SD + probiotic + PS); prebiotic smoking (PreS) (SD + prebiotic + PS), and symbiotic smoking (SymS)(SD + prebiotic + probiotic + PS). The animals were euthanized after 189 days of the experimental protocol. Results showed that supplementation with probiotics, prebiotics, and symbiotics significantly improved (P<0.05) the parameters: P, Ca, Mg, BMD, BMC, strength, resilience, and size of area of the femoral diaphysis of the animals chronically exposed or not cigarette smoke. We concluded that functional food supplementation improved the bone health of rats chronically exposed or not to cigarette smoke.

RESUMO: A fumaça de cigarro em grandes centros é uma das causas mais importantes de doenças inflamatórias crônicas em saúde pública e esta associada à diminuição de massa óssea, processo de consolidação e remodelação óssea. Os alimentos funcionais suplementados na dieta, devido sua capacidade de melhorar a absorção intestinal e competir com microrganismos patógenos, podem contribuir para a melhora da qualidade óssea. Portanto, o objetivo desse estudo foi avaliar os efeitos da suplementação de alimentos funcionais, probiótico, prebiótico ou simbiótico, na composição mineral, histomorfometria e nas propriedades biomecânicas ósseas de ratos em fase de crescimento expostos cronicamente a fumaça do cigarro (T). Sessenta e quatro ratos machos jovens foram randomicamente distribuídos em oito grupos (n=8): controle (C) [dieta basal (DB)]; probiótico (Pro) [DB + probiótico (Lactobacillus acidophilus, Enterecoccusfaecium, Bifidobacterium thermophilumandBifidobacterium longum (2-5 109 UFC cada)]; prebiótico (Pre) [DB + prebiótico (mananoligossacarídeo)]; simbiótico (Sym) (DB + probiótico + prebiótico); controle fumante (CS) [(DB + protocolo de exposição ao tabagismo passivo(PT)]; probiótico fumante (ProS) (DB + probiótico + PT); prebiótico fumante (PreS) (DB + prebiótico + PT); e simbiótico fumante (SymS) (DB + prebiótico + probiótico + PT). Os animais foram mortos após 189 dias de período experimental.Os resultados revelaram que a suplementação com probióticos, prebióticos e simbióticos melhoraram significativamente (P<0,05) os parâmetros: P, Ca, Mg, DMO, CMO, resistência, resiliência e tamanho da área das diáfises dos fêmures dos animais expostos, cronicamente ou não, a fumaça do cigarro. Os resultados permitem concluir que a suplementação dos alimentos funcionais melhorou a saúde óssea de ratos expostos cronicamente ou não a fumaça do cigarro.

Efeito da exposição à hidroquinona na resposta imune adaptativa induzida pela vacina contra a influenza / Effects of hydroquinone exposure on the adaptive immune response induced by the influenza vaccine

A gripe é causada pelo vírus Influenza e é um problema de saúde pública mundial, que pode levar a problemas sérios em idosos e crianças. O Brasil implantou a vacinação anual contra influenza a partir de 1999, como ação preventiva contra a doença. A vacina é produzida pelo Instituto Butantan e contém três cepas diferentes do vírus Influenza fragmentado para induzir resposta imune adaptativa, com produção de anticorpos específicos e neutralizantes. A literatura tem mostrado que a exposição à xenobióticos com potencial imunossupressor pode comprometer a eficácia de imunizações ativas, como a imunização contra a gripe. Nosso grupo de pesquisa tem mostrado que a exposição à hidroquinona (HQ), um composto tóxico presente em altas concentrações na fumaça do cigarro, prejudica a resposta imune inata e adquirida. Assim, este trabalho avaliou o efeito da exposição à HQ sobre a resposta imune à vacinação contra influenza. Camundongos machos da linhagem C57BL/6 foram diariamente expostos à HQ (2500 ppm) ou PBS, por 1 hora, por nebulização, por um período de 8 semanas. Durante este período, foram imunizados nas semanas 6 e 8 do início das exposições, pela injeção i.m. de 100µL da vacina. Os parâmetros tóxicos e imunológicos foram avaliados 7, 35 e 70 dias após a segunda dose da vacina. A exposição à HQ não alterou o peso corpóreo dos animais e nem causou alterações morfológicas no pulmão, fígado e rins (histologia por H&E); reduziu a frequência de hemácias (11%), hematócrito (14%), hemoglobina (14%) e volume celular (4%); causou estresse oxidativo no baço (citometria de fluxo); aumentou a área dos folículos de células B no baço e linfonodomegalia (histologia por H&E). Em conjunto, os dados aqui obtidos mostram que a exposição à HQ afetou mecanismos envolvidos na gênese da imunidade ativa contra influenza. Assim, os dados deste trabalho mostram mecanismos tóxicos ainda não descritos para a HQ, e ressalta a HQ como um poluente ambiental que deve ser considerado nas avaliações de risco

The flu is a health problem worldwide which is caused by the Influenza virus and may result in severe illness in infants and the elderly. The annually vaccination against influenza was implemented in Brazil in 1999 as a preventive measure. The vaccine is produced by Butantan Institute and contains three different strains of the inactivated Influenza virus which induce the adaptive immune response along with production of specific and neutralizing antibodies. The literature has shown that exposure to immunosuppressive xenobiotics may compromise the efficacy of active immunizations, such as influenza. Our research group has shown that exposure to hydroquinone (HQ), a toxic constituent of cigarette smoke, impairs both innate and adaptive immune response. Thus, the aim of this work was to evaluate the effects of HQ on the immune response induced by the influenza vaccine. Male C57BL/6 mice were daily exposed to HQ (2500 ppm) or PBS by nebulization, for 1 hour, for 8 weeks. During the exposure period, the animals were vaccinated on weeks 6 and 8 with 100µL of the vaccine. Toxicologic and immunological parameters were assessed 7, 35 and 70 days after boost administration. HQ exposure did not alter body weight and did not cause morphological alterations in the lungs, liver and kidneys (H&E staining); reduced the frequency of erythrocytes (11%), hematocrit (14%), hemoglobin (14%) and cellular volume (4%) and caused oxidative stress on the spleen (Flow Cytometry); increased the area of B cell follicles in the spleen and increased the size of draining lymph nodes (H&E staining). Altogether, these data show that HQ exposure affected mechanisms involved in the genesis of the adaptive immune response. Thus, the data presented in this work show toxic mechanisms of HQ that have not yet been described, and it also points out HQ as an environmental pollutant which should be considered on risk assessments

Protective effect of omega 3 and vitamin D in preventing lung-tissue damage of newborn rats exposed to cigarette smoke during intrauterine period / Efecto protector de omega 3 y vitamina D en la prevención del daño en el tejido pulmonar de ratas recién nacidas expuestas al humo del cigarrillo durante el período intrauterino

SUMMARY: This study aimed to investigate the toxic effects of cigarette smoke exposure on lung and the protective role of Omega 3 and Vitamin D against these toxic effects biochemically and histologically. 28 pregnant Wistar Albino rats were divided into four groups. The first group was control group; the second group was exposed to smoke of 10 cigarette by puff device 2 hours/day after pregnancy; the third group was exposed to cigarette smoke together with Omega 3 (0.5 mg/kg/day) and the fourth group was exposed to cigarette smoke together with vitamin D (42 microgram/kg/day). Finally, lung tissue sections of the newborn rats were stained with Hemotoxilen eosine and Masson tricromite. Malondialdehyde (MDA) and Fluorescent Oxidation Products (FOU) levels were measured. Fetal weights and the number of fetuses were significantly lower in the group received only cigarette smoke (both p<0.001). Histopathologically, pulmonary volume, number of developed alveols and parenchyma elasticity decreased significantly, meanwhile interstitial tissue increased, elastin and collagen did not develop adequately. Histopathologic changes significantly decreased in the group given Omega 3 and Vitamin D. Statistically, MDA and FOU levels were found to be higher in the group exposed to cigarette smoke compared to the control group, and MDA and FOU levels were lower in the group given Omega 3 along with cigarette smoke (p<0.001). Cigarette smoke caused histologically significant damage to fetal lung tissue, oxidative stress and increased MDA and FOU levels. This damage was significantly reduced with Omega 3 and Vitamine D supplementation. Omega 3 is an important antioxidant; vitamin D has no significant antioxidant effect.

RESUMEN: Este estudio tuvo como objetivo investigar los efectos tóxicos de la exposición al humo de cigarrillo en el pulmón, y el papel protector de Omega 3 y la Vitamina D contra esos efectos. 28 ratas Wistar albino preñadas fueron separadas en cuatro grupos. El primer grupo grupo control; el segundo grupo estuvo expuesto al humo de 10 cigarrillos por dispositivo de inhalación 2 horas / día después de la preñez; el tercer grupo se expuso al humo del cigarrillo junto con Omega 3 (0,5 mg / kg / día) y el cuarto grupo se expuso al humo del cigarrillo junto con vitamina D (42 microgramos / kg / día). Secciones de tejido pulmonar de las ratas recién nacidas se tiñeron con Hematoxilina Eosina y tricrómico de Masson. Se midieron los niveles de malondialdehído (MDA) y productos de oxidación fluorescente (POF). Los pesos fetales y el número de fetos fueron significativamente más bajos en el grupo que recibió solamente humo de cigarrillo (ambos p <0,001). Histopatológicamente, el volumen pulmonar, el número de alveolos desarrollados y la elasticidad del parénquima disminuyeron significativamente; mientras que el tejido intersticial aumentó y la elastina y el colágeno no se desarrollaron adecuadamente. Los cambios histopatológicos disminuyeron significativamente en el grupo que recibió Omega 3 y Vitamina D. Estadísticamente, se encontró que los niveles de MDA y POF eran más altos en el grupo expuesto al humo de cigarrillo en comparación con el grupo control, además los niveles de MDA y POF fueron más bajos en el grupo que recibió Omega 3 junto con el humo del cigarrillo (p <0,001). El humo del cigarrillo causó daños histológicamente significativos en el tejido pulmonar fetal, el estrés oxidativo y el aumento de los niveles de MDA y FOU. Este daño se redujo significativamente con los suplementos de Omega 3 y Vitamina D. El omega 3 es un importante antioxidante; la vitamina D no tiene ningún efecto antioxidante significativo.

Screening and bioinformatics analysis of specific microRNAs in testes of rats exposed to cigarette smoke

Objective: To identify effects of cigarette smoke on the male reproductive capacity and to explore the miRNA expression in the testes after cigarette smoke exposure. Methods: Eighty male rats were conducted by factorial analysis of variance designed for cigarette exposure. A microarray was employed to detect the differential expression of miRNA in the testis tissue of smoke-exposed rats. Results: Four miRNAs (miR-138-5p, miR-181d-5p, miR-19a-3p, and miR-3588) were significantly downregulated and one miRNA (miR-155-5p) was upregulated in the testes of smoke-exposed rats compared with control rats. This result was further confirmed by a quantitative RT-PCR assay, and pathological changes were observed in the testes. Bioinformatics analysis showed that the predicted target genes were closely related to the regulation of the apoptosis pathway. Conclusions: miRNA may play an important role in the smoke-exposure-induced testicular toxicity of male rats.

Mrp1 is up-regulated in lung tissues of mildly stable COPD mice induced by smoke, Nrf2 pathway may be involved / 中国药理学通报

Aim: To investigate the effect of Nrf2 pathway on the expression of MRP1 in mildly stable COPD mice. Methods: The mild COPD mouse model was established by passive cigarette smoking. The pathological changes of lung tissues were examined by HE staining. Immunohistochemistry and Western blot were used to detect the protein expression of MRP1, Nrf2 and HO-1. Results: Compared with normal group, each lung function index of the mild-moderate COPD model group was significantly lower, but compared with wide type(WT) model group, the reduction was more significant in Nrf2-/- model group. HE results showed diffuse inflammatory reaction and alveolar bronchial structure damage in alveolar of WT and Nrf2-/- model mice, and it was more pronounced in Nrf2-/- mice. Immunohistochemistry and Western blot results showed that the expression of MRP1 in lung tissue of Nrf2-/- normal mice was significantly reduced compared with the normal WT group. After passive cigarette smoking, The expression of MRP1, Nrf2 and HO-1 in WT model group increased significantly, but compared with Nrf2-/- normal mice, there was no significant change in the expression of MRP1 in Nrf2-/- model group. Conclusions: Mildly stable COPD mice may counteract the xenobiotic damage caused by cigarette smoke through up-regulating the expression of MRP1 protein, which may be associated with Nrf2 signaling activation.

Protective effect of water extract of guibi-tang against pulmonary inflammation induced by cigarette smoke and lipopolysaccharide / 한국실험동물학회지

Water extract of guibi-tang (GB), a traditional Chinese, Japanese, and Korean herbal medicine, is used to treat memory impairment, insomnia, and peptic ulcers. The aim of this study was to investigate the protective effects of GB on pulmonary inflammation induced by cigarette smoke (CS) and lipopolysaccharide (LPS). C57BL/6 mice were used to develop a pulmonary inflammation model by exposing them to CS for 1 h per day for 7 days. LPS was intranasally administered to mice under mild anesthesia on day 5. GB was administered 1 h before CS exposure at doses of 50 or 100 mg/kg for 7 days. Our results showed that GB suppressed the CS and LPS induced elevation in inflammatory cell counts in the bronchoalveolar lavage fluid (BALF), with significant reductions in protein, tumor necrosis factor (TNF)-α, and interleukin (IL)-6 levels. Histological studies revealed that GB decreased the inflammatory cell infiltration into lung tissue caused by CS- and LPS-exposure. GB also significantly decreased the CS and LPS-induced expression of inducible nitric oxide synthase (iNOS) in the lung tissue. Taken together, GB effectively attenuated airway inflammation caused by CS and LPS. These results indicate that GB is a potential therapeutic herbal formula for pulmonary inflammatory disease.

Pharmacological approach to restoring steroid sensitivity in respiratory diseases / 中国药理学与毒理学杂志

Inhaled corticosteroid is the first-line controller for asthma and COPD. However, about 10% of the asthmatics (severe/refractory asthma) and most of the COPD patients are resistant to the beneficial effects of corticosteroids.There is a pressing unmet medical need to develop novel therapeu-tic agents to restore corticosteroid efficacy in affected patients. There have been reports showing the promise of theophylline and rapamycin in reversing steroid resistance in COPD. Our laboratory has demonstrated that andrographolide, a bioactive diterpenoid lactone isolated from the plant Androgra-phis paniculata, is an effective anti-inflammatory and anti-oxidative compound in both asthma and COPD experimental models. In a severe asthma mouse model using combined IFN-γ/LPS exposure, production of IL-27 and methacholine-induced airway hyperresponsiveness (AHR) were found to be corticosteroid-resistant.Andrographolide was found to restore the anti-inflammatory effect of dexameth-asone in LPS/IFN-γ-induced IL-27 levels in bronchoalveolar lavage(BAL)fluid and AHR in mice.LPS/IFN-γ markedly reduced the nuclear level of histone deacetylase-2 (HDAC2), an essential epigenetic enzyme that mediates corticosteroid anti-inflammatory actions. Andrographolide significantly restored nuclear HDAC2 levels and diminished total HAT/HDAC activity ratio in mouse lungs exposed to LPS/IFN-γ, probably via suppression of PI3K/Akt/HDAC2 phosphorylation and up-regulation of the antioxi-dant transcription factor Nrf2 level. In a cigarette smoke (CS)-induced COPD model, andrographolide markedly restored dexamethasone actions in inhibiting CS-induced lung neutrophilia.In addition,androgra-pholide facilitated dexamethasone actions to suppress BAL fluid IL-6, IL-1b, KC and IL-17 levels. In lung lysates, andrographolide markedly restored total nuclear HDAC activity. The complete steroid re-sensitization mechanism of andrographolide remains to be unraveled. Nevertheless, our existing data strongly implicate a potentially novel steroid re-sensitizing activity of andrographolide in both severe asthma and COPD models.