Effect of cigarette smoke extract on the polymorphonuclear leukocytes chemiluminescence: influence of a filter containing glutathione.

Cigarette smoking is known to be a risk factor for several chronic and neoplastic diseases. Many compounds formed by cigarette burning, ranging from particulate materials to water solutes and gaseous extracts, are considered to be noxious agents, and many biochemical and molecular mechanisms have been proposed for the toxic effects of cigarette smoke. The oral cavity and the upper respiratory tract represent the first contact areas for smoke compounds; even a single cigarette can produce marked effects on some components of the oral cavity, either chemical compounds, such as glutathione and enzymes, or cellular elements, such as polymorphonuclear leukocytes. Several studies suggest a protective role of glutathione against the noxious effects of tobacco smoke; the sulphydril groups of glutathione, in fact, could react with some smoke products, such as unsaturated aldehydes, leading to the formation of harmless intermediate compounds and simultaneously preventing the inactivation of metabolically essential molecules, such as some enzymes. In this paper we analyse the effect of a filter containing glutathione on the respiratory burst of polymorphonuclear leukocytes exposed to aqueous extract of cigarette smoke, measuring their chemiluminescence activity. The results of this paper indicate that the GSH-containing filter has a likely protective effect against the inhibition of cigarette smoke extract on polymorphonuclear leukocyte activity.     

香烟烟雾水溶液中的活性氧及其对姊妹染色单体互换的影响

SCE频率分析是研究环境诱变剂和致癌剂引起人类遗传物质损伤的有效手段。以人类外周血林巴细胞姊妹染色单体交换（SCE）为指标,评价了香烟烟雾水溶液的遗传毒性。首先,定量测定了香烟烟雾水溶液中含有H2O2、O2-等活性氧,从而间接证明了其潜在的遗传毒性。然后,将一定量的香烟烟雾水溶液加入人外周血淋巴细胞培养液中,结果表明实验组SCE频率明显高于空白对照组,因此直接证明了香烟烟雾水溶液的遗传毒性。     

Analysis of Hydrogen Peroxide in an Aqueous Extract of Cigarette Smoke and Effect of pH on the Yield

An analysis of hydrogen peroxide in an aqueous extract of cigarette smoke, which contains many redox-active compounds, requires a method with high selectivity. An aqueous extract of the particulate phase of cigarette smoke was analyzed by HPLC with an electrochemical detector (ECD). Samples were prepared by collecting the particulate phase of the cigarette smoke on a glass fiber filter and extracting it with a phosphate buffer. The obtained solution was purified by using a Waters Oasis MCX cation-exchange cartridge, and then analyzed by an HPLC-ECD system with a Shodex KS-801 mixed-mode resin column. Pre-injecting hydrogen peroxide at a high concentration into the HPLC instrument stabilized the analytical results. The recovery of hydrogen peroxide by using an extract of the particulate phase of the cigarette smoke was more than 80%. An increase in the amount of hydrogen peroxide was observed during extraction with the phosphate buffer at higher pH values. In contrast, extraction with phosphoric acid did not increase the amount of hydrogen peroxide during extraction.     

Cigarette smoke-induced pulmonary inflammation is attenuated in CD69-deficient mice

Cluster of differentiation 69 (CD69) has been identified as a lymphocyte early activation marker, and recent studies have indicated that CD69 mediates intracellular signals and plays an important role in various inflammatory diseases. Cigarette smoke (CS) is a strong proinflammatory stimulus that induces the release of proinflammatory mediators by recruiting macrophages and neutrophils into the lung tissue, and is one of the main risk factors for a number of chronic diseases. However, the potential role of CD69 in CS-induced pulmonary inflammation has not been determined. To address to this question, CD69-deficient (KO) and wild-type (WT) mice were subjected to CS-induced acute pulmonary inflammation. After the exposure with CS, the expression of CD69 in the lung of WT mice was significantly induced, it was predominantly observed in macrophages. In conjunction with this phenomenon, neutrophil and macrophage cell counts, and expression of several cytokines were significantly higher in the bronchoalveolar lavage fluid (BALF) of CS-exposed WT mice compared with air-exposed WT mice. Likewise, the CS-induced accumulation of inflammatory cells and cytokines expression were significantly lower in CD69-KO mice than in WT mice. These results suggest that CD69 on macrophages is involved in CS-induced acute pulmonary inflammation.     

烤烟主流烟气内源有害成分与烟叶化学成分相关性

研究了不同生态产区烟叶主要化学成分与主流烟气内源有害成分的关联性.结果表明:初烤烟叶内源有害成分和危害性指数(H值)与烟叶部位具有较好的关联性.4-甲基亚硝基-1-(3-吡啶基)-1-丁酮(NNK)、苯并[a]芘、氢氰酸(HCN)、氨和苯酚的释放量表现为上部叶＞中部叶,巴豆醛释放量表现为中部叶＞上部叶,H值表现为上部叶＞中部叶.不同内源有害成分与烟叶含碳、含氮化学成分的相关性复杂,同一类型烟叶化学成分对不同内源有害成分呈现出不同的相关关系.卷烟危害性指数与烟叶中烟碱、蛋白质、总氮、主要多酚和有机酸含量呈显著正相关,而与钾离子、总糖、还原糖、淀粉等含碳类物质含量呈显著负相关.适当降低烟叶含氮化合物含量、增加烟叶钾含量可能会降低烟叶的危害性指数.     

Evolutionary origins of taste buds: phylogenetic analysis of purinergic neurotransmission in epithelial chemosensors

Taste buds are gustatory endorgans which use an uncommon purinergic signalling system to transmit information to afferent gustatory nerve fibres. In mammals, ATP is a crucial neurotransmitter released by the taste cells to activate the afferent nerve fibres. Taste buds in mammals display a characteristic, highly specific ecto-ATPase (NTPDase2) activity, suggesting a role in inactivation of the neurotransmitter. The purpose of this study was to test whether the presence of markers of purinergic signalling characterize taste buds in anamniote vertebrates and to test whether similar purinergic systems are employed by other exteroceptive chemosensory systems. The species examined include several teleosts, elasmobranchs, lampreys and hagfish, the last of which lacks vertebrate-type taste buds. For comparison, Schreiner organs of hagfish and solitary chemosensory cells (SCCs) of teleosts, both of which are epidermal chemosensory end organs, were also examined because they might be evolutionarily related to taste buds. Ecto-ATPase activity was evident in elongate cells in all fish taste buds, including teleosts, elasmobranchs and lampreys. Neither SCCs nor Schreiner organs show specific ecto-ATPase activity, suggesting that purinergic signalling is not crucial in those systems as it is for taste buds. These findings suggest that the taste system did not originate from SCCs but arose independently in early vertebrates.     

Curcumin Combats Against Cigarette Smoke and Ethanol-Induced Lipid Alterationsin Rat Lung and Liver

Background: Human population, in spite of the medical and scientific achievements, still fall as a prey to the evils of habitual smoking and alcohol, thus necessitating safer counteracting measures. Objective: To evaluate the effect of cotreatment of curcumin (Curcuma longa) in rats subjected to acute exposure to cigarette smoke (CS) and ethanol (EtOH). Methodology: Of the four groups of experimental rats, a set of rats was subjected to whole body exposure to cigarette smoke along with ethanol administration serving as a model of CS+EtOH injury. Curcumin treatment was given to two sets of rats: (i) one set receiving simultaneous CS+EtOH and (ii) one set of normal rats without any administration. The other group of rats served as control. Blood, liver and lung of rats were selected for assessment of CS+EtOH injury as well as curcumin treatment. Result: Altered lipid, lipoprotein profile and bile acid excretion were observed in CS+EtOH rats along with premalignant pathological state in tissues. In treated rats, the levels were maintained at near-normal levels along with near-normal histology. Conclusion: This biochemical picture on cotreatment with curcumin suggests that curcumin could counteract the injurious effects of combined CS and EtOH and thus might help to reduce the risk of hyperlipidemic disorders which develop due to smoking and drinking.     

茶多酚单体L-EGCG对气相烟引起鼠肺细胞膜损伤的抑制作用

茶多酚(GTP)单体L-EGCG对香烟气相物质损伤鼠肺细胞膜的保护作用研究结果表明,L-EGCG能抑制香烟气相物质引发的脂质过氧化;用脂肪酸自旋标记物5-DOXYL和16-DOXYL标记鼠肺细胞膜,发现预先加入L-EGCG可抑制气相烟引起的膜浅层流动性改变,并与L-EGCG的浓度呈量效关系。在0.001到0.1mg/mL浓度范围内,L-EGCG本身对膜的浅层没有影响,而能使膜深层的流动性略有增大。由试验推测,L-EGCG的保护作用很可能是由于清除了气相烟中的自由基或脂质过氧化产生的脂类自由基。     

Effects of cigarette smoke on degranulation and NO production by mast cells and epithelial cells

Exhaled nitric oxide (eNO) is decreased by cigarette smoking. The hypothesis that oxides of nitrogen (NO_X) in cigarette smoke solution (CSS) may exert a negative feedback mechanism upon NO release from epithelial (AEC, A549, and NHTBE) and basophilic cells (RBL-2H3) was tested in vitro. CSS inhibited both NO production and degranulation (measured as release of beta-hexosaminidase) in a dose-dependent manner from RBL-2H3 cells. Inhibition of NO production by CSS in AEC, A549, and NHTBE cells was also dose-dependent. In addition, CSS decreased expression of NOS mRNA and protein expression. The addition of NO inhibitors and scavengers did not, however, reverse the effects of CSS, nor did a NO donor (SNP) or nicotine mimic CSS. N-acetyl-cysteine, partially reversed the inhibition of beta-hexosaminidase release suggesting CSS may act via oxidative free radicals. Thus, some of the inhibitory effects of CSS appear to be via oxidative free radicals rather than a NO_X -related negative feedback.     

Cigarette smoke exposure induces ROS-mediated autophagy by regulating sestrin,AMPK, and mTOR level in mice

Many pathological conditions linked to cigarette smoking are caused by the production of reactive oxygen species (ROS). The present study was conducted to analyze the effect of ROS on the lungs of Swiss mice exposed to cigarette smoking, focusing on autophagy-mediated mechanisms, and investigate the involvement of SESN2, AMPK, and mTOR signaling. Mice were exposed to cigarette smoke (CS) for 7, 15, 30, 45, and 60 days; the control group was not exposed to CS. Only mice exposed to CS for 45 days were selected for subsequent N-acetylcysteine (NAC) supplementation and smoke cessation analyses. Exposure to CS increased the production of ROS and induced molecular changes in the autophagy pathway, including an increase in phosphorylated AMPK and ULK1, reduction in phosphorylated mTOR, and increases in SESN2, ATG12, and LC3B levels. NAC supplementation reduced ROS levels and reversed all molecular changes observed upon CS treatment, suggesting the involvement of oxidative stress in inducing autophagy upon CS exposure. When exposure to CS was stopped, there were decreases in the levels of oxidative stress, AMPK and ULK1 phosphorylation, and autophagy-initiating molecules and increase in mTOR phosphorylation. In conclusion, these results suggest the involvement of ROS, SESN2, AMPK, and mTOR in the CS-induced autophagic process in the lung.     

茶多酚单体L-EGCG对气相烟引起鼠肺细胞膜损伤的抑制作用

茶多酚(GTP)单体L-EGCG对香烟气相物质损伤鼠肺细胞膜的保护作用研究结果表明,L-EGCG能抑制香烟气相物质引发的脂质过氧化;用脂肪酸自旋标记物5-DOXYL和16-DOXYL标记鼠肺细胞膜,发现预先加入L-EGCG可抑制气相烟引起的膜浅层流动性改变,并与L-EGCG的浓度呈量效关系。在0.001到0.1mg/mL浓度范围内,L-EGCG本身对膜的浅层没有影响,而能使膜深层的流动性略有增大。由试验推测,L-EGCG的保护作用很可能是由于清除了气相烟中的自由基或脂质过氧化产生的脂类自由基。