谷胱甘肽对香烟烟雾作用大鼠淋巴细胞氧化应激的影响

谷胱甘肽是由谷氨酸、半胱氨酸和甘氨酸组成的三肽类巯基物质,以2种形式存在:还原型(GSH)和氧化型(GSSG)。GSH与许多重要的生物学现象,如基因表达调控DNA生物合成、酶活力和代谢调节、免疫功能调节代谢等密切相关,作为细胞内重要的抗氧化剂和解毒剂,对保护细胞、延缓衰老有重要     

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。     

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     

你知道香烟烟雾的去向吗

大家都知道吸烟是有害的,但烟草是如何侵犯我们的身体,大多数人可能并不了解。这根白色的小棍里除了尼古丁,还有焦油、一氧化碳等上百种"生化武器"在密谋着一起颠覆健康的演变。未清洗的"抽油烟机"     

锌7-金属硫蛋白对严重烫伤大鼠肝脏氧化应激反应的作用

AIM: Using the model of burned animal with delayed resuscitation to study antagonistic effect of Zn7 - metallothionein(Zn7-MT) on oxidative stress in the liver of rats suffered from severe thermal injury on skin. METHODS: Tocompare the changes in antioxidant concentrations and antioxidative enzyme activities in the liver or plasma ofburned rats with or without Zn7-MT in resuscitation fluid by biochemical assay. RESULTS: After injury, glutathione concentration was progressively decreased with time, At24 h after injury, activities of glutathione reductase and glutathione peroxidase in the liver of burned rats were increased and then decreased at 48 h postburn, αTocopherol in plasma was reduced at 24 h and malondialdehyde in the liver was increased significantly postburn.MT and MT-1 mRNA expression in burned rats were activated. Taken together, oxidative stress in the liver ofburned rats occurred. Exogenous Zn7-MT attenuated the changes in antioxidant concentrations and antioxidativeenzyme activities in the liver or plasma of burned rats. The effect of Zn7-MT was in a concentration-dependentmanner and the concentration of 10μtmol/L was the most effective. Exogenous Zn7-MT also inhibited MT-1 mRNA overexpression and increased MT protein concentration. CONCLUSION: Zn7-MT effectively antagonizedoxidative stress in the liver of rats with severe thermal injury.     

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。     

Oxidative damage in keratinocytes exposed to cigarette smoke and aldehydes

Cigarette smoke (CS) is a significant environmental source of human exposure to chemically active saturated (acetaldehyde) and α,β-unsaturated aldehydes (acrolein) inducing protein carbonylation and dysfunction. The exposure of oral tissues to environmental hazards is immense, especially in smokers. The objectives of the current study were to examine the effect of aldehydes originating from CS on intracellular proteins of oral keratinocytes and to observe the antioxidant response in these cells.Intracellular protein carbonyl modification under CS, acrolein and acetaldehyde exposure in the HaCaT keratinocyte cell line, representing oral keratinocytes was examined by Western blot. Possible intracellular enzymatic dysfunction under the above conditions was examined by lactate dehydrogenase (LDH) activity assay. Oxidative stress response was investigated, by DCF (2,7-dichlorodihydrofluorescein) assay and GSH (glutathione) oxidation.Intracellular protein carbonyls increased 5.2 times after CS exposure and 2.7 times after exposure to 1 μmol of acrolein. DCF assay revealed an increase of fluorescence intensity 3.2 and 3.1 times after CS and acrolein exposure, respectively. CS caused a 72.5% decrease in intracellular GSH levels compared to controls. Activity of intracellular LDH was preserved.α,β-Unsaturated aldehydes from CS are capable of intracellular protein carbonylation and have a role in intracellular oxidative stress elevation in keratinocytes, probably due to the reduction in GSH levels.     

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.     

Oxidative stress and innate immunity responses in cigarette smoke stimulated nasal epithelial cells

Cigarette smoke extracts (CSE) may play a significant role in diseases of the upper airway including chronic rhinosinusitis. Even short term exposure of cigarette smoke has adverse effects on mitochondrial functions and redox homeostasis in tissues which may progress to further complications associated with chronic smoking. Cigarette smoke alters toll-like receptor 4 (TLR4) expression and activation in bronchial epithelial cells. Carbocysteine is an anti-oxidant and mucolytic agent. The effects of carbocysteine on CSE induced oxidative stress and on associated innate immune and inflammatory responses in nasal epithelial cells are largely unknown. The present study was aimed to assess in CSE stimulated nasal epithelial cells (RPMI 2650) the effects of carbocysteine (10⁻⁴ M) on: cell survival, intracellular reactive oxygen species (ROS) production, TLR4 expression, LPS binding and neutrophil chemotaxis (actin reorganization). We found that CSE increased ROS production, TLR4 expression, LPS binding and neutrophil chemotaxis and all these events were counteracted by pre-incubating CSE stimulated RPMI 2650 cells with carbocysteine. In conclusion, the present study provides compelling evidence that carbocysteine may be considered a promising therapeutic strategy in chronic inflammatory nasal diseases.     

Oxidative stress and oxidative damage in chemical carcinogenesis

Reactive oxygen species (ROS) are induced through a variety of endogenous and exogenous sources. Overwhelming of antioxidant and DNA repair mechanisms in the cell by ROS may result in oxidative stress and oxidative damage to the cell. This resulting oxidative stress can damage critical cellular macromolecules and/or modulate gene expression pathways. Cancer induction by chemical and physical agents involves a multi-step process. This process includes multiple molecular and cellular events to transform a normal cell to a malignant neoplastic cell. Oxidative damage resulting from ROS generation can participate in all stages of the cancer process. An association of ROS generation and human cancer induction has been shown. It appears that oxidative stress may both cause as well as modify the cancer process. Recently association between polymorphisms in oxidative DNA repair genes and antioxidant genes (single nucleotide polymorphisms) and human cancer susceptibility has been shown.     

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.     

Studying the impact of S9 on cyto-genotoxicity of cigarette smoke in human peripheral blood lymphocytes in vitro

In present study, human lymphocytes were exposed to cigarette smoke condensates (CSCs) at the doses of 25, 50, 75, 100 and 125 μg ml⁻¹ with and without S9, and the cyto-genotoxic effects were detected with CCK-8, cell apoptosis and micronucleus assays. DNA repair kinetics was observed with comet assay. Our results indicated that the cell viability decreased with CSCs doses, the percentages of apoptosis cell and the frequencies of micronuclei increased with CSCs doses, and DNA damage of human lymphocytes induced by CSCs could be basically repaired within 240 min. However, the cytotoxicity induced by CSCs +S9 was significantly lower than that induced by CSCs −S9 in CCK-8 and cell apoptosis assays, and the DNA repair speed in +S9 group was quicker than that in −S9 group. In conclusion, S9 may affect not only the cyto-genotoxicity of CSCs but also the repair process of DNA damage induced by CSCs in lymphocytes.     

Age dependent differential effects of cigarette smoke on hepatic and pulmonary xenobiotic metabolizing enzymes in rats

The effects of cigarette smoke (CS) on hepatic and pulmonary monooxygenase (MO) activities (aniline␣4-hydroxylase, AH; aminopyrine N-demethylase, AMND; 7-ethoxyresorufin O-deethylase, EROD; p-nitroanisole O-demethylase, p-NAOD), lipid peroxidation (LP), and reduced glutathione (GSH) levels and glutathione S-transferase (GST) activities toward several substrates (1-chloro-2,4-dinitrobenzene, CDNB; 1,2-dichloro-4-nitrobenzene, DCNB; ethacrynic acid, EAA; 1,2-epoxy-3-( p-nitrophenoxy)-propane, ENPP) were determined in 20-, 90- and 360-day-old male rats. The animals were exposed to CS five times a day, with 1 h intervals, for 3 days in a chamber supplied alternatively with smoke and fresh air, and were killed 16 h after the last treatments. The hepatic AH activity increased significantly in 20-day-old rats and remained unaltered in older age groups. The hepatic AMND activity unaltered, significantly increased and decreased in 20-, 90- and 360-day-old rats, respectively. The pulmonary AH activity increased significantly in 20- and 90-day-old rats whereas no alteration was noted in 360-day-old rats. CS was ineffective on pulmonary AMND activity at all ages. CS increased hepatic and pulmonary EROD and p-NAOD activities significantly in all age groups compared to controls. In liver, LP level was significantly increased, decreased, and unaltered in 20-, 90- and 360-day-old rats, respectively. CS increased hepatic GSH level significantly in 90-day-old rats but was not effective in the other age groups. In lung, LP level was increased in 90- and 360-day-old rats and unaltered in 20-day-old rats. CS increased pulmonary GSH level significantly in 90-day-old rats and did not have any effect in the other age groups. The hepatic GST activities toward CDNB and DCNB decreased significantly in 360-day-old rats and were unaltered in the younger age groups. The hepatic GST activity toward EAA was unaltered, significantly increased and decreased in 20-, 90- and 360-day-old rats, respectively. The hepatic GST activity toward ENPP decreased significantly in 20- and 90-day-old rats but was unaltered in the oldest group of rats. In 20-day-old rats, the pulmonary GST activity toward ENPP increased significantly whereas the other GST activities did not alter. In 90-day-old rats, however, CS significantly decreased all the pulmonary GST activities studied. Unaltered DCNB GST, significant increase in EAA GST and decrease in CDNB and ENPP GST activities of lung were noted in 360-day-old rats. These results reveal that the regulation in rats of hepatic and pulmonary MO and GST activities are differentially influenced by CS as a function of age. Received: 28 January 1997 / Accepted: 7 May 1997     

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.     

Systemic effects of acute cigarette smoke exposure in mice

Abstract

Context: Cigarette smoke (CS) causes both pulmonary and extrapulmonary disorders.

Objective: To determine the pulmonary and extrapulmonary effects of acute CS exposure in regard to inflammation, oxidative stress and DNA damage.

Materials and methods: Mice were exposed to CS for 10 days and then their lungs, heart, liver, pancreas, kidneys, gastrocnemius muscle and subcutaneous (inguinal and flank) and visceral (retroperitoneum and periuterus) adipose tissues were excised. Bronchoalveolar lavage fluid samples were obtained for differential cell analysis. Inflammatory cell infiltration of the tissues was assessed by immunohistochemistry for Mac-3⁺ cells, F4/80⁺ cells and CD45⁺ cells. Oxidative stress was determined by immunohistochemistry for thymidine glycol (a marker of DNA peroxidation) and 4-hydroxy hexenal (a marker of lipid peroxidation), by enzyme-linked immunosorbent assay for protein carbonyls (a marker of protein peroxidation) and by measurements of enzyme activities of glutathione peroxidase, superoxide dismutase and catalase. DNA double-strand breaks were assessed by immunohistochemistry for γH2AX.

Results: CS exposure-induced inflammatory cell infiltration, oxidative stress and DNA damage in the lung. Neither inflammatory cell infiltration nor DNA damage was observed in any extrapulmonary organs. However, oxidative stress was increased in the heart and inguinal adipose tissue.

Discussions: Induction of inflammatory cell infiltration and DNA damage by acute CS exposure was confined to the lung. However, an increased oxidative burden occurred in the heart and some adipose tissue, as well as in the lung.

Conclusions: Although extrapulmonary effects of CS are relatively modest compared with the pulmonary effects, some extrapulmonary organs are vulnerable to CS-induced oxidative stress.     

丙烯腈对大鼠脑皮层氧化应激效应的研究

目的　评价长期低浓度接触AN对大鼠脑组织的氧化性应激效应 ,探讨AN神经毒性机制。方法　SD雄性大鼠 3 0只 ,随机分成 3组 :对照组、低剂量组和高剂量组 ,通过饮水分别给予 0、5 0和 2 0 0mg L的AN溶液。染毒时间为 12周。染毒结束后从每组随机选取 7只大鼠 ,取双侧大脑皮层测定氧化性应激指标。结果　染毒组的MDA水平(nmol mg蛋白 )明显高与对照组 (16 60± 6 49) ,且高剂量组 (15 7 7± 85 40 )明显高于低剂量组 (98 5 3± 5 0 3 5 ) ,其剂量 效应关系有显著性 ;3组之间的GSH Px活力差异未见显著性 ;虽然低剂量组的SOD活力和对照组的差异未见显著性 ,但高剂量组明显高于对照组和低剂量组。高剂量组GSH明显低于对照组和低剂量组 ,但低剂量组和对照组的差异无显著性。结论　诱导氧化性应激可能是AN神经毒性的重要作用机制。