Scholarly article on topic 'Short-Term Systemic Effects of Nose-Only Cigarette Smoke Exposure in Mice: Role of Oxidative Stress'

Short-Term Systemic Effects of Nose-Only Cigarette Smoke Exposure in Mice: Role of Oxidative Stress Academic research paper on "Environmental engineering"

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Academic research paper on topic "Short-Term Systemic Effects of Nose-Only Cigarette Smoke Exposure in Mice: Role of Oxidative Stress"

Cellular Physiology Cell Physiol Biochem 2013;31:15-24

, DOI: 10.1159/000343345

and Biochemistry Published online: January 14, 2013

© 2013 S. Karger AG, Basel www.karger.com/cpb

Accepted: October 24,2012

1421-9778/13/0311-0015$38.00/0

Original Paper

Karger Open access

This is an Open Access article licensed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License (www.karger.com/OA-license), applicable to the online version of the article only. Distribution for non-commercial purposes only.

Short-Term Systemic Effects of Nose-Only Cigarette Smoke Exposure in Mice: Role of Oxidative Stress

Abderrahim Nemmara Haider Razab Deepa Subramaniyana Javed Yasind Annie Johnb Badreldin H. Alic Elsadig E. Kazzamd

aDepartment of Physiology, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, UAE; bDepartment of Biochemistry, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, UAE; cDepartment of Pharmacology and Clinical Pharmacy, College of Medicine & Health Sciences, Sultan Qaboos University, Al-Khod, Sultanate of Oman; dDepartment of Internal Medicine, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, UAE

Key Words

Cigarette smoke • Nose-only exposure, short-term exposure • Systolic blood pressure • Thrombosis • Liver enzymes • Oxidative stress

Abstract

Background/Aims: Long-term cigarette smoking (CS) is a major risk factor for respiratory and cardiovascular diseases, and is also known to adversely affect other organs. However, data on the systemic effects of short-term CS exposure (STCSE) are scarce. Presently, using a nose-only exposure system, we evaluated the systemic effects of STCSE in mice. Methods: We assessed the effects of CS generated by 9 consecutive cigarettes per day for 4 days in a nose-only exposure system on cardiovascular, hepatic and renal endpoints evaluated on day 5 in mice. Control mice were exposed to air only. Results: CS significantly increased systolic blood pressure and decreased total nitric oxide plasma concentration. Circulating platelets and erythrocyte numbers were also increased. However, STCSE did not significantly increase thrombosis in pial arterioles and venules. STCSE significantly raised plasma alanine aminotransferase and gamma glutamyl transpeptidase activities, but did not affect urea or creatinine concentrations. Interestingly, while STCSE enhanced the production of reactive oxygen species in heart and kidney and lipid peroxidation in heart, liver and kidneys, it also enhanced the antioxidant activity of superoxide dismutase, probably indicating that STCSE causes adaptive reactions to counterbalance the potentially damaging action of oxygen radicals induced by STCSE. Conclusion: These results suggest that STCSE causes blood pressure increase, hepatotoxicity and oxidative stress in the heart, liver and the kidneys. These data provide information on the initial steps leading to the systemic effects of STCSE, a stage at which the diseases may likely be reversed.

Copyright © 2013 S. Karger AG, Basel ^

Prof. A. Nemmar United Arab Emirates University, College of Medicine and Health Sciences 9

Department of Physiology, P.O. Box 17666, Al Ain (United Arab Emirates) ^^

Tel. +971-37137533, Fax +971 3 7671966 V E-Mail anemmar@uaeu.ac.ae, anemmar@hotmail.com

Cellular Physiology Ce" phYsio1 Biochem 2013;31:15-24

, DOI: 10.1159/000343345 I© 2013 S. Karger AG, Basel and Biochemistry Published online: January 14, 2013_[www.karger.com/cpb__16

Nemmar/Raza/Subramaniyan et al.: Short-Term Systemic Effects of Cigarette Smoke

Introduction

Cigarette smoke (CS) is a major risk factor for chronic obstructive pulmonary disease (COPD), lung cancer, and cardiovascular diseases. COPD is characterized by an excessive inflammatory response in the airways, parenchyma and pulmonary vasculature [1, 2].

In addition to the typical pulmonary changes in COPD, several extrapulmonary effects have been recognized [1, 2]. Several studies have reported the occurrence of systemic events following CS exposure including increase in plasma concentrations of proinflammatory cytokines and oxidative stress [1, 2]. Moreover, it has been recognized that systemic features and other diseases are more common in COPD, including skeletal muscle dysfunction, cardiovascular disease, and diabetes, all of which are thought to have an analogous inflammation-based mechanisms to COPD [1].

Even in the absence of COPD, CS alone can cause significant extrapulmonary diseases such as coronary artery disease. Young smokers or even passive smokers may show endothelial dysfunction of the systemic vessels and systemic oxidative stress [3, 4]. Oxidative stress causes muscle fatigue and facilitates proteolysis [5].

Extensive evidence has been reported on the pulmonary and systemic effects of chronic CS exposure. Such experiments require chronic exposure to CS lasting for several months [1, 6]. However, there is a paucity of data on the short-term effects of CS. The later can give specific information on the initial changes in the pulmonary and extrapulmonary sites, and will likely offer the possibility to reverse them.

We have recently demonstrated that short-term nose-only exposure to CS, a system that best resembles the human exposure situation, induced increase in airway resistance, pulmonary inflammation and morphological changes, and oxidative stress in the lung tissue [7]. Several studies investigated the effect of CS on oxidative stress in various organs including skeletal muscle, lung, liver, kidney or bladder muscle [8-10]. However, as far as we are aware, no study has comprehensively assessed the short-term nose-only effects of CS on cardiovascular system (blood pressure, circulatory cells and thrombosis), liver and kidney functions and the role of oxidative stress thereon. Therefore, the specific aim of this study is to investigate the systemic effects of short-term nose-only CS exposure on systolic blood pressure, pial microvessel thrombosis, liver and kidney functions, and the effect on oxidative stress comprising the concentrations of reactive oxygen species and lipid peroxidation and superoxide dismutase activity in the heart, liver and kidney.

Materials and Methods

Animals and treatments

This project was reviewed and approved by the Institutional Review Board of the United Arab Emirates University, Faculty of Medicine and Health Sciences, and experiments were performed in accordance with protocols approved by the Institutional Animal Care and Research Advisory Committee (8/4/2012, protocol No. A7-12,).

CS exposure

BALB/C mice (Taconic Farms Inc., Germantown, NY, USA) were housed in a conventional animal house and maintained on a 12-hour light-dark cycle. The animals were placed in cages and supplied with pelleted food and water ad libitum. Following an acclimatization period of one week, animals were randomly divided into two groups: control (air) and CS-exposed. Mice were placed in soft restraints and connected to the exposure tower. Animals were exposed to mainstream CS generated by commercially available filtered cigarettes (Marlboro red, 12 mg tar/1.0 mg nicotine; Philip Morris, Richmond, VA) through their noses using a nose-only exposure system (InExpose System, Scireq, Canada). A computer-controlled puff was generated every minute, leading to 10 s of CS exposure followed by 50 s of fresh air. CS-exposed group inhaled CS from 9 consecutive cigarettes per day for 4 days as previously described [7, 11]. Control animals were treated similarly but were exposed to filtered air for the same duration. The total particulate density concentration

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Cellular Physiology Cell Physiol Biochem 2013;31:15-24

, DOI: 10.1159/000343345 I© 2013 S. Karger AG, Basel and Biochemistry Published online: January 14, 2013_[www.karger.com/cpb__17

Nemmar/Raza/Subramaniyan et al.: Shirt-Term Systemic Effects if Cigarette Smoke

of CS was measured daily, and indicated an average of 420.5 mg total particulate matter per m3 (TPM/m3) in the tower [7].

Systolic blood pressure (SBP) measurement

Following the exposure to CS, the systolic BP (SBP) was measured using a computerized noninvasive tail-cuff manometry system (ADInstrument, Colorado Springs, USA). To avoid procedure-induced anxiety, mice were trained for 5 consecutive days before the experimental procedure [12].

Blood collection and analysis

The same animals used to measure SBP were anesthetized with intraperitoneal injection of sodium pentobarbital (45 mg/kg), and blood was drawn from the inferior vena cava in EDTA (4 %). A sample was used for platelets and white blood cells (WBC) counts using an ABX VET ABC Hematology Analyzer with a mouse card (ABX Diagnostics, Montpellier, France). The remaining blood was centrifuged at 4°C for 15 min at 900 g and the plasma samples were stored at -80°C until further analysis.

The determination of nitric oxide (NO) was performed with a total NO assay kit from R & D systems (Minneapolis, MN, USA) which measures the more stable NO metabolites NO2- and NO3-[13, 14].

The alanine aminotransferase (ALT), aspartate transaminase (AST), gamma-glutamyl transpeptidase (GGT) activities, and urea and creatinine concentrations were measured using standard laboratory methods with LX20 multiple automated analyser (Beckman Coulter, CA, USA).

Measurement of oxidative stress, lipid peroxidation and superoxide dismutase (SOD) activity in heart,

liver and kidneys

Following the exposure to CS or air, animals were sacrificed by an overdose of sodium pentobarbital, and their heart, liver and kidney tissues were quickly collected and rinsed with ice-cold PBS (pH 7.4) before homogenization in 0.1M phosphate buffer pH 7.4 containing 0.15M KCl, 0.1mM EDTA, 1mM DTT and 0.1mM phenylmethylsulfonylfluoride at 4oC. Homogenates were centrifuged for 10 min at 3000xg to remove cellular debris and supernatants were used for further analysis. Protein content was measured by Bradford's method as described before [15, 16].

Measurement of ROS and lipid peroxidation: ROS in heart, liver and kidney tissues of all mice were measured using 2', 7'- Dichlorofluorescein diacetate (DCFDA; Molecular Probes, Eugene, OR, USA) as a fluorescent probe as described before [17, 18]. NADPH-dependent membrane lipid peroxidation was measured as thiobarbituric acid reactive substance using malonedialdehyde as standard (Sigma-Aldrich Fine Chemicals, St Louis, MO, USA) [18].

SOD activity was measured as the conversion of NBT to NBT-diformazan according to the vendor's protocol (R & D System, MN, USA). The extent of reduction in the appearance of NBT-formazan was used as a measure of SOD activity present in the tissues.

Experimental pial arterioles and venules thrombosis model

In a separate experiment, in vivo thrombogenesis in the pial arterioles and venules was assessed after CS exposure, according to a previously described technique [19, 20]. Briefly, the trachea was intubated after induction of anaesthesia with urethane (1mg/g body weight, i.p.), and a 2F venous catheter (Portex, Hythe, UK) was inserted in the right jugular vein for the administration of fluorescein (Sigma, St. Louis, MO, USA). After that, a craniotomy was first performed on the left side, using a microdrill, and the dura was stripped open. Only untraumatized preparations were used, and those showing trauma to either microvessels or underlying brain tissue were discarded. The animals were then placed on the stage of a fluorescence microscope (Olympus, Melville, NY, USA) attached to a camera and DVD recorder. A heating mat was placed under the mice and body temperature was raised to 37°C, as monitored by a rectal thermoprobe connected to a temperature reader (Physitemp Instruments, NJ, USA). The cranial preparation was moistened continuously with artificial cerebrospinal fluid of the following composition (mM): NaCl 124, KCl 5, NaH2PO4 3, CaCl2 2.5, MgSO4.4, NaHCO3 23 and glucose 10, pH 7.3-7.4. A field containing arterioles and venules 1520 |im in diameter was chosen. Such a field was taped prior to, and during the photochemical insult, which was carried out by injecting fluorescein (0.1ml/mouse of 5% solution) via the jugular vein, which was allowed to circulate for 30-40 sec. The cranial preparation was then exposed to stabilized mercury light. The combination produces endothelium injury of the arterioles. This, in turn, causes platelets to adhere

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Cellular Physiology Ce" phYsio1 Biochem 2013;31:15-24

, DOI: 10.1159/000343345 I© 2013 S. Karger AG, Basel and Biochemistry Published online: January 14, 2013_[www.karger.com/cpb__18

Nemmar/Raza/Subramaniyan et al.: Short-Term Systemic Effects of Cigarette Smoke

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Fig. 1. Systolic blood pressure (A), and total nitric oxide (NO) levels in plasma (B) following short-term nose-only cigarette smoke exposure or air (control). Data are mean ± SEM (n = 8).

Fig. 3. Thrombotic occlusion time in pial arterioles (A) and venules (B) following short-term nose-only cigarette smoke or air (control) exposure. Data are mean ± SEM (n = 8).

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Fig. 2. Leukocyte (A), platelet (B) and erythrocyte (C) numbers following short-term nose-only cigarette smoke or air (control) exposure. Data are mean ± SEM (n = 8).

Fig. 4. Gamma-glutamyl transpeptidase (A), alanine aminotranferease (B) and aspartate transaminase (C) activities in plasma following short-term nose-only cigarette smoke or air (control) exposure. Data are mean ± SEM (n = 8).

at the site of endothelial damage and then aggregate. Platelet aggregates and thrombus formation grow in size until complete vascular occlusion. The time from the photochemical injury until full vascular occlusion (time to flow stop) in arterioles were measured in seconds. At the end of the experiments, the animals were euthanized by an overdose of urethane.

Statistics

All statistical analyses were performed with GraphPad Prism Software version 4. (San Diego, CA, USA). •

To determine whether parameters were normally distributed, the Kolmogorov-Smirnov statistic normality

Cellular Physiology Ce" phYsio1 Biochem 2013;31:15-24

, DOI: 10.1159/000343345 I© 2013 S. Karger AG, Basel and Biochemistry Published online: January 14, 2013_[www.karger.com/cpb__19

Nemmar/Raza/Subramaniyan et al.: Short-Term Systemic Effects of Cigarette Smoke

Fig. 5. Urea (A) and creatinine (B) in plasma following short-term nose-only cigarette smoke or air (control) exposure. Data are mean ± SEM (n = 8).

Air Cigarette smoke

Fig. 6. Reactive oxygen species in the heart (A), liver (B) and kidney (C) tissues following short-term nose-only cigarette smoke or air (control) exposure. Data are mean ± SD (n=8).

test was applied. Normally distributed data were analyzed using the unpaired t-test for differences between groups whereas non-normally ones where analyzed with Mann Whitney test. P-values <0.05 were considered as significant. All the data in figures were reported as mean ± SEM.

Results

Systolic blood pressure and NO levels in plasma

Short-term exposure to CS caused a significant increase in SBP compared to air-exposed mice (Fig. 1A). The NO (which is known to diffuse into vascular smooth muscle cells and causes vasorelaxation) levels have significantly decreased in plasma of mice exposed to CS compared to air-exposed mice (Fig. 1B).

Leukocyte, platelet and erythrocyte numbers in blood

Compared to control group, short-term nose-only CS exposure caused a significant increase in platelet (P<0.0001) and erythrocyte (P<0.01) numbers (Fig. 2B-C). However, the number of leukocytes was not significantly affected by CS exposure (Fig. 2A).

Thrombosis in pial venules and arterioles

The assessment of thrombosis in pial arterioles and venules revealed the absence of prothrombotic effect of CS at the studied time point. Figure 3 shows that compared to airexposed mice, the thrombotic occlusion time in arterioles and venules were not affected • following short-term exposure to CS.

Cellular Physiology Ce" phYsio1 Biochem 2013;31:15-24

, DOI: 10.1159/000343345 I© 2013 S. Karger AG, Basel and Biochemistry Published online: January 14, 2013_[www.karger.com/cpb__20

Nemmar/Raza/Subramaniyan et al.: Short-Term Systemic Effects of Cigarette Smoke

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Fig. 7. Lipid peroxidation in the heart (A), liver (B) and kidney (C) tissues following short-term nose-only cigarette smoke or air (control) exposure. Data are mean ± SD (n=8).

Fig. 8. Superoxide dismutase in the heart (A), liver (B) and kidney (C) tissues following short-term nose-only cigarette smoke or air (control) exposure. Data are mean ± SD (n=8).

GGT ALT and AST in plasma

Short-term nose-only exposure to CS caused a significant increase in the activity of GGT (P<0.005) and ALT (P=0.01) compared to control group (Fig. 4A-B). The AST activity was slightly but insignificantly increased following short-term exposure to CS.

Urea and Creatinine in plasma

Figure 5 illustrates the effect of CS on plasma concentrations of urea and creatinine. Following short-term nose-only CS exposure, the plasma concentrations of urea and creatinine were not increased significantly compared to air-exposed mice.

Reactive oxygen species concentrations in heart, liver and kidney tissuesShort-term nose-only CS exposure induced a significant increase in the concentration of reactive oxygen species in heart (P<0.0001) and kidney (P<0.0005) tissues compared to air exposed group (Fig. 6A and C). In the liver, the observed increase of reactive oxygen species concentration did not reach statistical significance (Fig. 6B).

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is depicted in Figure 7. Compared to control group, a significant increase in lipid peroxidation were observed in the heart (p<0.05; Fig. 7A), liver (P<0.0001; Fig. 7B) and kidney (P=0.0005; Fig. 7C) following short-term nose-only exposure to CS.

Cellular Physiology Ce" phYsio1 Biochem 2013;31:15-24

, DOI: 10.1159/000343345 I© 2013 S. Karger AG, Basel and Biochemistry Published online: January 14, 2013_[www.karger.com/cpb__21

Nemmar/Raza/Subramaniyan et al.: Short-Term Systemic Effects of Cigarette Smoke

SOD activity in the heart, liver and kidney tissues

Figure 8 illustrates the effect of short-term nose-only CS exposure on the concentrations of the antioxidant SOD in heart, liver and kidney tissues. CS exposure caused a significant increase of SOD activity in the heart (P<0.0005, Fig. 8A), liver (P<0.001, Fig. 8B) and kidney (P<0.005, Fig. 8C) tissues compared to their respective control group.

Discussion

The present work provides evidence that short-term nose-only exposure to CS induced an increase in systolic blood, platelet and erythrocyte numbers and tissue-specific enzymes including AST and GGT but without causing thrombotic events in pial venules or arterioles. Moreover, markers of oxidative stress were all increased in the heart, liver and kidney tissues of mice exposed to CS.

While substantial information has been reported on the systemic effects of chronic smoke exposure, there is a paucity of data on the short-term systemic effects of smoking. One of the reasons why it is important to know these effects is because repetitive short-term smoke effects may constitute the underlying causal chain of reactions leading to the ultimate chronic effects. Therefore, studying the short-term effect of CS can give more specific information and reflect the initial changes in the pathophysiological mechanisms of CS-induced chronic systemic effects.

Several exposure systems are being used to study the effect CS exposure in mice [8, 21, 22]. The smoking machines that have been employed with animal models comprise systems that use nose-only or whole body exposures. The shortcoming of using whole body exposure is that the animals may ingest nicotine or tar substances when cleaning their fur. The nose-only exposure system avoids this problem and most likely best resembles the human situation [8, 21, 22]. Using the nose-only exposure system, we have recently demonstrated CS exposure causes an increase in airway resistance, pulmonary inflammation and morphological changes, and oxidative stress in the lung tissue [7]. The advantage of the current study is that we assessed the extrapulmonary effects of short-term nose-only exposure to CS on various set of indices comprising SBP, circulatory cells, pial thrombosis, liver and kidney functions and the oxidative stress in the heart, liver and kidney.

It has been reported that chronic exposure CS exposure can increase systemic oxidative stress [1], alter NO bioavailability [23], cause endothelial dysfunction [24, 25], and influence the levels of other major risk factors, such as blood pressure [26]. Our data show that short-term exposure to CS induces a significant increase in SBP. Along with that, we observed a significant decrease in the concentration of total NO in plasma following short-term exposure to CS. Our data obtained on day 5 post-CS exposure are in line with those of Guo et al. [27] who reported that short-term (6 weeks) and long-term (16 weeks) CS exposure cause increase in arterial pressure and a marked decrease in NO metabolite. They also reported a correlation between NO and the change of structural and mechanical status of arterial wall in response to CS [27].

We found that short-term exposure to CS causes a significant increase in platelet and erythrocyte numbers. This finding is suggestive of a rapid bone marrow response, causing an increase in RBC and platelet numbers. Recently, it was reported that acute exposure to nanoparticles and particulate air pollution caused a rapid elevation in platelet counts [28, 29]. Tamagawa et al. [28] found that the intensity of the bone marrow response correlated with the amount of particles phagocytosed by alveolar macrophages in the lung, indicating a strong link between lung and systemic events. The increase in RBCs that we observed in our study is in line with the finding of Kung et al. [30] who reported an increase of RBC and haematocrit in young men smokers. Furthermore, it is well established that higher red blood cell counts, hematocrit, blood viscosity, and an ongoing inflammatory process potentiate the prothrombotic process associated with smoke exposure [31]. •

Cellular Physiology Ce" phYsio1 Biochem 2013;31:15-24

, DOI: 10.1159/000343345 I© 2013 S. Karger AG, Basel and Biochemistry Published online: January 14, 2013_[www.karger.com/cpb__22

Nemmar/Raza/Subramaniyan et al.: Short-Term Systemic Effects of Cigarette Smoke

The prothrombotic effects of exposure to CS have been repeatedly demonstrated before [31]. It has been reported that CS exposure causes alterations in platelet function, antithrombotic/prothrombotic factors, and fibrinolytic factors [31]. The lack of prothrombotic effects in pial arterioles and venules observed in our study suggest that short-term exposure to CS does not cause prothrombotic effects. Dong et al. [32] demonstrated that the effects of CS on thrombosis require a proatherosclerotic background. Indeed, they reported that the exposure to CS (4h/day, 5 days/week for 12 weeks) causes prothrombotic effects in the carotid artery of ApoE-/- mice but not in wild-type C57Bl/6 mice. The later displayed similar fibrinogen binding and thrombotic occlusion time similar to air-exposed mice [32].

Our data show a significant increase of GGT and ALT, indicating that CS causes hepatotoxicity. This finding points up that the liver might be a potential target organ for short-term CS toxity. Increases in liver enzymes were reported in young men (age range, 18.6-22.8 yr; mean age, 19.4 yr) exposed to CS and were closely correlated with number of cigarettes smoked daily [30]. The absence of a signifcant effect on urea and creatinine plasma concentration by short-term exposure to CS suggests lack of a gross insult to the kidneys, but does not necessarily rule out an adverse effect on renal function, as it has been shown that the classical kidney function tests such as the urea and creatinine may not detect subtle renal insults [33].

There is strong evidence showing a consistent association between cigarette smoking and increase in the mortality rate from smoking-related diseases such as pulmonary and cardiovascular diseases [2]. The role of free radicals in the pathogenesis of these diseases has been well documented [2, 34]. Formation of free radicals and ROS is a normal consequence of a variety of biochemical reactions. However, these free radicals can cause oxidative damage to the tissues through lipid peroxidation. The human body has defence mechanism, which comprise free radical scavenger enzymes namely superoxide dismutase, catalase and glutathione peroxidase [6, 35]. To further assess the mechanism underlying the extrapulmonary effects of short-term exposure to CS, we have assessed the ROS and lipid peroxidation in heart, liver and kidneys. Our findings show that short-term exposure to CS cause oxidative stress in the heart, liver and kidneys. The mechanisms behind these extrapulmonary effects remain to be investigated but could result from the inflammation and oxidative stress taking place in the lung that result in overspill in circulation causing systemic inflammation and oxidative stress affecting various organs [1]. Moreover, our data show a significant increase of the activity of the antioxidant enzyme SOD in the heart, liver and kidneys. This indicates that the development of oxidative stress is accompanied by an adaptive response that counterbalances the potentially damaging activity of oxygen free radicals by antioxidant defence mechanisms. We and others have reported an increase pulmonary oxidative stress following short-term exposure to CS [7, 36] . It has been shown that immediately following exposure to mainstream CS by nose-only inhalation (at varying doses 40, 120, 240 puffs), dose-dependent decreases in pulmonary and renal GSH were observed in rats whereas, in guinea pigs, reductions in pulmonary, hepatic and renal GSH were observed only at the highest level of exposure [9]. Others reported no effects of nose-only CS exposure on GSH content in muscle or lung homogenates in guinea pig lungs 3h and 24h following nose-only CS exposure [8]. Inhalation of CS for 30 days, three times a day in rats, resulted in a significant decrease of the total free glutathione contents in the lung and liver but not in the heart and kidney [10]. Also, elevated concentrations of oxidized glutathione and protein S-thiolation were observed in the lung but not in other tissues [10]. These discrepancies could be explained by species differences in metabolizing the components of CS, protocol of exposure, duration of exposure or other unknown factors. Further studies are required to clarify this issue.

In conclusion, short-term exposure to CS induced increase in systolic blood pressure, platelet and erythrocyte numbers and liver enzymes including AST and GGT but without causing thrombotic events in pial venules or arterioles. Moreover, CS exposure induces oxidative stress in the heart, liver and kidney tissues of mice. Further work is needed to elucidate the celluar and molecualr mechanims underlying these effects.

Cellular Physiology Cell Physiol Biochem 2013;31:15-24

, DOI: 10.1159/000343345 I© 2013 S. Karger AG, Basel and Biochemistry Published online: January 14, 2013_[www.karger.com/cpb__23

Nemmar/Raza/Subramaniyan et al.: Short-Term Systemic Effects of Cigarette Smoke

Acknowledgements

This work was supported by the UAEU, UAEU-SQU, CMHS and Terry Fox grants. The authors would like to thank Mrs S. Zia for her technical help.

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Cellular Physiology and Biochemistry

Cell Physiol Biochem 2013;31:15-24

DOI: 10.1159/000343345 Published online: January 14, 2013

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Nemmar/Raza/Subramaniyan et al.: Short-Term Systemic Effects of Cigarette Smoke

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