香烟烟雾溶液作用下大鼠淋巴细胞的氧化应激反应

目的　以细胞内活性氧自由基 (ROS)水平、DNA损伤、脂质过氧化物 (LPO)水平以及超氧化物歧化酶 (SOD)活力为指标 ,研究细胞在香烟烟雾溶液作用下产生的氧化应激反应。方法　以PBS作吸收液 ,用大包氏管收集香烟主流烟雾 ,将香烟烟雾溶液 (CSS)分别以 0、1× 10 - 3、2× 10 - 3、4× 10 - 3、8× 10 - 3、12× 10 - 3、16× 10 - 3支 ml的浓度作用于大鼠淋巴细胞。用 2′ ,7′ 二乙酰二氯荧光素 (DCFH DA)测定细胞内ROS水平 ,用彗星试验检测细胞DNA损伤 ,同时检测细胞内SOD活力和LPO水平。结果　 2× 10 - 3支 ml以上剂量组二氯荧光素 (DCF)荧光强度、彗星尾长以及LPO水平均显著高于对照组 ,且随剂量增加而增加。 16× 10 - 3支 ml组SOD活力显著低于对照组。与对照组相比 ,1× 10 - 3支 ml组LPO水平显著降低、SOD活力则显著增高。结论　CSS达到一定剂量时使细胞内ROS水平增高 ,引起细胞DNA损伤和脂质过氧化 ,高剂量时SOD活力降低 ,而低剂量时能够诱导SOD活力     

香烟烟雾对巨噬细胞的氧化损伤及盐酸氨溴索抗氧化作用的研究

目的研究香烟烟雾对体外培养的巨噬细胞的氧化损伤及盐酸氨溴索的抗氧化作用。方法采用黄嘌呤氧化酶法及硫代巴比妥酸法测定香烟烟雾提取物(CSE)和CSE+盐酸氨溴索作用于体外培养的小鼠巨噬细胞后30min、1、3和6h超氧化物歧化酶(SOD)和脂质过氧化物(LPO)的含量。结果加入CSE后3h,CSE组巨噬细胞SOD活性明显低于空白对照组,差异有统计学意义(P<0.05);CSE+氨溴索组SOD活性明显高于CSE组,差异有统计学意义(P<0.05);其余时间点三组SOD活性差异均无统计学意义。加入CSE后1h,CSE组巨噬细胞LPO含量明显高于空白对照组,差异有统计学意义(P<0.05);CSE+氨溴索组LPO含量与CSE组及空白对照组比较差异均无统计学意义;其余时间点三组LPO含量差异均无统计学意义。结论香烟烟雾作用于巨噬细胞产生脂质过氧化反应,造成氧化性损伤。盐酸氨溴索具有抗氧化作用,在一定程度上抑制过氧化反应,减轻氧化反应所致的损伤。     

香烟烟雾与慢性应激联合作用对大鼠生殖功能的影响

目的 研究香烟烟雾与慢性应激联合作用对大鼠生殖功能的影响.方法 健康清洁级SD大鼠40只,雌雄各半,随机分为对照组、香烟烟雾组、慢性应激组和联合作用组,每组10只.对照组不采取任何处理;香烟烟雾组大鼠进行呼吸道静式染毒,每次10支香烟,1次/d,每次30 min;慢性应激组从5种慢性应激方案中每日随机给予一种慢性应激,...     

香烟烟雾对家兔的毒害作用

通过对家兔被动吸烟及静脉注射香烟烟雾溶液的毒害作用试验,推算出主动吸烟的人香烟中毒量;并通过对其病理解剖及组织观察,见到香烟烟雾能引起如下变化:使细胞的脂质膜溶掉,切断体内细胞的生物氧化链,血红蛋白变性,心肌疏离,心肌内血管收缩,以及使肺、肝等器官产生较多的癌变前期的细胞。     

黄芩苷对香烟烟雾提取物诱导的细胞损伤保护作用研究

目的研究黄芩苷对香烟烟雾提取物（cigarette smoke extract,CSE）诱导的人肺泡上皮细胞损伤的影响,并初步探讨其机制。方法制备CSE,用不同浓度黄芩苷对人肺泡上皮细胞A549进行预处理,再用CSE刺激细胞。用MTT法检测细胞存活率,流式细胞术测定细胞凋亡率,彗星实验观察DNA损伤情况,荧光法测定细胞内活性氧（reactive oxygen species,ROS）含量。结果随着CSE浓度的增加和作用时间的延长,细胞存活率下降,各组之间有统计学差异（P〈0.05）;黄芩苷能减少CSE诱导的细胞存活率下降,抑制CSE诱导的细胞内ROS的产生,并有剂量依赖关系（P〈0.05或P〈0.01）;黄芩苷＋CSE组细胞的彗星尾长、尾部DNA含量、尾距、Olive尾距均小于CSE组（P〈0.05）,并且黄芩苷可以减少CSE导致的细胞凋亡。结论黄芩苷可以拮抗CSE对细胞的损伤,提高细胞的存活率,降低凋亡率,其原因可能与黄芩苷能降低细胞ROS含量,减少DNA损伤有关。     

香烟烟对支气管哮喘的影响

香烟烟对支气管哮喘的影响山西医科大学第一医院（０３０００１）王秀清王莲芸阳泉矿务局医院王怀云香烟烟是人们在工作和日常生产中不可能完全避免的物质，国内外对吸烟与变应性疾病关系的报道较少。我们试图通过对香烟烟与支气管哮喘发病关系的探讨，解释在不同的年龄和...     

香烟烟雾凝聚物对人胚肾类上皮细胞转化作用的研究

用10和15μg/ml的香烟烟雾凝聚物处理人胚肾原代细胞24小时,一个月左右细胞开始发生转化。转化细胞的超微结构出现一些特征性的改变:细胞的染色体数目变化明显;通过H~3-TdR的掺入量来看,DNA合成旺盛;细胞能在半固体琼脂上生长,目前体外传代培养巳在4年以上,即已获得不死性。     

BDT滤嘴对香烟烟雾凝聚物诱发微核率的降低作用

用双核微核法测定香烟烟雾凝聚物对ＢＡＬＢ／ｃ－３Ｔ３和ＣＨＬ细胞的致微核作用，结果发现香烟烟雾凝聚物可使两种细胞的微核率升高，不需Ｓ９的活化，比较不同滤嘴对香烟烟雾凝聚物中诱变物质的清除效果，发现普通滤嘴不能有效地降低微核率，而ＢＤＴ滤嘴则能使微核率降低了一半左右。     

香烟烟雾提取物致小鼠生殖细胞的遗传学损伤及其机制

吸烟与许多疾病有关，但其对遗传或生殖损害研究报道尚少。香烟烟雾提取物可致小鼠生殖细胞染色体损伤，但该作用是否可能遗传给后代或影响生殖细胞的功能尚未见报道。香烟烟雾中含有许多自由基，而自由基可致DNA损伤。我们给雄性小鼠腹腔注射不同剂量香烟烟雾提取物，观察生殖细胞染色体     

Prenatal exposure to cigarette smoke enhances oxidative stress in astrocytes of neonatal rat

Oxidative stress is involved in the pathogenesis of smoking-related disease. Protection of astrocytes from oxidative insult appears essential to maintain brain function. In this study, we have investigated the effect of gestational cigarette exposure on astrocyte survival. Pregnant female were randomly allocated to the control group or to the cigarette smoke group in which they were placed in an exposure chamber and inhale three cigarettes smoke twice a day for a period of 20 days. The control group was kept in the exposure chamber for the same duration, but without exposure to cigarette smoke. Newborn rats from both groups were weighed 24?h after birth and then cerebral hemispheres were collected for astrocyte culture. Incubation of astrocytes isolated from animals exposed to cigarette smoke with 300?μM H₂O₂ for 1?h induced a significant decrease of the proportion of surviving cells compared to cells isolated form control animals. We have observed that H₂O₂-treated astroglial cells derived from cigarette smoke exposure showed more reduced superoxide dismutase and catalase activities than H₂O₂-treated astroglial cells from control animals. In conclusion, this study indicates that astroglial cells derived from newborn rats exposed in utero to cigarette smoke are more vulnerable to oxidative assault than cultured astrocytes obtained from control animals. These results point out the existence of excitotoxic lesions in newborn exposed in utero to cigarette smoke and suggest that despite their high antioxidative activities, astrocytes cannot survive and protect neurons under massive oxidative stress.     

Immunohistochemical evaluation of oxidative stress in murine lungs after cigarette smoke exposure

Cigarette smoking generates an oxidative stress in the lung, which may contribute to the pathogenesis of chronic obstructive pulmonary disease. We performed an immunohistochemical study to evaluate oxidative stress in the lung after acute cigarette smoke (CS) exposure in mice. Paraffin-embedded lung tissue sections were prepared from mice exposed and unexposed to CS for 1 h. The sections were immunostained with antibodies against 8-hydroxy-2'-deoxyguanosine (8-OHdG), an oxidative DNA adduct, and 4-hydroxy-2-nonenal (4-HNE), a lipid peroxidation product. The bronchiolar and alveolar epithelium of mice unexposed to CS exhibited weak signals for 8-OHdG and 4-HNE, whereas by 1 h after CS exposure the signals in the bronchiolar epithelial cells and the alveolar epithelial cells, particularly type II cells, had increased dramatically. The increases in both were associated with increased 8-OHdG levels in bronchoalveolar lavage fluid as determined by enzyme-linked immunoassay. These results suggest that acute CS exposure imposes oxidative stress predominantly on bronchiolar epithelial and alveolar type II cells, confirming that cigarette smoking causes oxidative damage to the respiratory epithelium.     

Diphenyl diselenide prevents oxidative damage induced by cigarette smoke exposure in lung of rat pups

The effect of cigarette smoke exposure on lungs of rat pups was evaluated. Animals were exposed to passive cigarette smoke during 3 weeks and a number of toxicological parameters in lung of pups were examined, such as lipid peroxidation, δ-aminolevulic acid dehydratase (δ-ALA-D) activity, components of the enzymatic antioxidant defenses (superoxide dismutase (SOD) and catalase activities) and non-enzymatic antioxidant defenses (Vitamin C and non-protein thiol (NPSH) levels). Furthermore, a possible protective effect of diphenyl diselenide, (PhSe)₂, was studied. The results demonstrated an increase in lipid peroxidation, an inhibition of δ-ALA-D activity, a reduction of Vitamin C and NPSH levels induced by cigarette smoke exposure, indicating damage in lungs of rat pups. Oral administration of (PhSe)₂ (0.5 mg/kg) restored TBARS levels, non-enzymatic antioxidant defenses and activity of δ-ALA-D. These results indicated that exposure to cigarette smoke enhanced oxidative stress, thereby disturbing the tissue defense system. (PhSe)₂ protected against oxidative damage induced by cigarette smoke exposure in lung of rat pups.     

Apoptotic effects of cigarette smoke extracts on mouse embryonic stem cells via oxidative stress

Previous studies have reported that cigarette smoke and cigarette smoke extract (CSE) have negative effects on embryonic development. However, no studies have investigated the mechanism through which CSE affects the cellular signaling pathway leading to apoptosis and oxidative stress in embryonic cells, or how the two pathways are cross‐linked. Thus, we studied the effects of CSE on apoptosis and oxidative stress in mouse embryonic stem cells (mESCs). Specifically, we measured changes in cell viability in response to CSEs (3R4F and two domestic cigarettes CSE 1 and 2) using a water soluble tetrazolium (WST) assay and a neutral red uptake (NRU) assay, which revealed that cell viability decreased in a concentration‐dependent manner. Western blot analysis revealed that the expression of cyclin D1 and cyclin E1 was decreased and that of p21 and p27 was increased by CSE. Additionally, the number of terminal deoxynucleotidyl transferase (TUNEL)‐stained cells was increased by CSE, while the levels of Bax and Caspase‐3 increased and Bcl‐2 decreased. Moreover, a 2′,7′‐dichlorofluorescin diacetate (DCF‐DA) assay and reactive oxygen species (ROS)‐Glo H₂O₂ assay confirmed that ROS were generated in response to CSE and that they were associated with up‐regulated Keaf‐1 and CHOP. Overall, the results revealed that cigarette smoke extract (CSE) inhibited cell proliferation by regulating cell cycle‐related protein expression and increased oxidative stress by regulating the expression of Kelch‐like ECH‐associated protein 1 (Keap‐1) and CCAAT/enhancer‐binding protein homologous protein (CHOP), resulting in apoptosis in mESCs.     

Evidence for cigarette smoke-induced oxidative stress in the rat pancreas

Background/aims: Recent findings with a rodent model of cigarette smoke inhalation revealed a causal relationship between chronic exposure to cigarette smoke and the development of pancreatitis. The present study was conducted to ascertain whether cigarette smoke induces oxidative stress in the rat pancreas concurrently with inflammation.

Methodology: Rats (six per treatment group) were treated for 0, 3, 6, 9, or 12 weeks with cigarette smoke (0.7?mg/L). Pancreatic tissues were examined for histological and pathological alterations and serum for changes in interleukin-6 concentration. Pancreatic expression and localization of α-smooth muscle actin, transforming growth factor-β1, and collagen-1 were determined as measures of progressive inflammation/fibrosis. Pancreatic superoxide dismutase and glutathione peroxidase activities and malondialdehyde content were measured as indices of oxidative stress.

Results: Inflammatory cell infiltration and ductal hyperplasia were detected in pancreata after 12 weeks of treatment with cigarette smoke. The serum interleukin-6 concentration increased significantly and pancreatic glutathione peroxidase activity declined significantly after 12 weeks of treatment. No other significant changes were observed.

Conclusions: Pancreata of rats exposed chronically to cigarette smoke exhibit inflammation concurrently with suppression of glutathione peroxidase activity. These observations favor a role for oxidative stress in the induction of pancreatitis associated with chronic cigarette smoke inhalation.     

Aryl hydrocarbon receptor protects lung adenocarcinoma cells against cigarette sidestream smoke particulates-induced oxidative stress

Environmental cigarette smoke has been suggested to promote lung adenocarcinoma progression through aryl hydrocarbon receptor (AhR)-signaled metabolism. However, whether AhR facilitates metabolic activation or detoxification in exposed adenocarcinoma cells remains ambiguous. To address this question, we have modified the expression level of AhR in two human lung adenocarcinoma cell lines and examined their response to an extract of cigarette sidestream smoke particulates (CSSP). We found that overexpression of AhR in the CL1-5 cell line reduced CSSP-induced ROS production and oxidative DNA damage, whereas knockdown of AhR expression increased ROS level in CSSP-exposed H1355 cells. Oxidative stress sensor Nrf2 and its target gene NQO1 were insensitive to AhR expression level and CSSP treatment in human lung adenocarcinoma cells. In contrast, induction of AhR expression concurrently increased mRNA expression of xenobiotic-metabolizing genes CYP1B1, UGT1A8, and UGT1A10 in a ligand-independent manner. It appeared that AhR accelerated xenobiotic clearing and diminished associated oxidative stress by coordinate regulation of a set of phase I and II metabolizing genes. However, the AhR-signaled protection could not shield cells from constant oxidative stress. Prolonged exposure to high concentrations of CSSP induced G0/G1 cell cycle arrest via the p53-p21-Rb1 signaling pathway. Despite no effect on DNA repair rate, AhR facilitated the recovery of cells from growth arrest when CSSP exposure ended. AhR-overexpressing lung adenocarcinoma cells exhibited an increased anchorage-dependent and independent proliferation when recovery from exposure. In summary, our data demonstrated that AhR protected lung adenocarcinoma cells against CSSP-induced oxidative stress and promoted post-exposure clonogenicity.     

Inhibition of salivary enzymes by cigarette smoke and the protective role of glutathione

Tobacco smoke is involved in the pathogenesis of several diseases regarding different body systems, mainly cardiovascular and respiratory in addition to its local toxic effect in the oral cavity. The noxious effects of smoke compounds justify the high incidence of periodontal diseases, caries, and neoplastic diseases of oral tissues in smokers. Some toxic components of tobacco smoke, unsaturated and saturated aldehydes, could interact with thiol rich compounds, leading to structural and functional modification of these molecules. Previous papers have demonstrated an in vitro significant decrease of some enzymatic activities, both in plasma and in saliva, following external addition of aldehydes or exposure to cigarette smoke (CS). Furthermore, the same studies underlined the protective effect exerted by the addition of glutathione (GSH) against the damaging role of smoke aldehydes. In this study some salivary enzymes (lactic dehydrogenase [LDH], aspartate aminotransferase [AST] and amylase), and total GSH were measured in 20 volunteers smokers, before and just after smoking a single cigarette. All enzymatic activities showed a significant inhibition following a single cigarette, probably due to the interaction between smoke aldehydes and -SH groups of the enzyme molecules. Moreover, the percentage of the enzymatic inhibition showed a negative correlation with the basal level of salivary GSH. Our results emphasize that not only one cigarette is sufficient to impair the salivary enzymatic activities but also strengthen the proposed protective role of GSH against the noxious biochemical effects of CS.     

Trichostatin A alleviated ovarian tissue damage caused by cigarette smoke exposure

Cigarette smoke (CS) has a negative impact on women's health and fertility. Studies have shown that histone deacetylases 1 and 2 (HDAC1/2) were involved in oocyte development. However, the roles of HDAC1/2 in ovarian toxicity caused by CS exposure and the therapeutic potential of trichostatin A (TSA, a HDAC inhibitor) for ovarian tissue damage have not been investigated. In this study, Female C57BL/6 mice were exposed to CS from six cigarettes mixed with indoor air for 120 min (one cigarette for 20 min) using a whole-body mainstream smoke exposure system twice daily for 30 days. TSA (0.6 mg/kg body weight) was injected intraperitoneally into mice in the Control + TSA group and CS + TSA group every two days for 30 days. We found that exposure to CS resulted in ovarian tissue damage and HDAC1/2 over-expression. TSA alleviated the structural changes of ovarian tissue induced by smoking and prevented the activation of HDAC1/2. Exposure to CS caused autophagy inhibition and pyroptosis activation. TSA treatment restored the expression of autophagy-associated proteins and decreased the levels of pyroptosis-related proteins induced by CS exposure. The TSA effect may be mediated by inhibition of HDAC1/2 involved in autophagy and pyroptosis process.