Oxidative Stress for Smoking Persons in Suburbs Mosul City *

The study was conducted in Mosul-suburb to show the effects of smoking on antioxidants and some biochemical parameters which include: Vitamin E, Vitamin A, βcarotene, Vitamin C, Folic acid, Ceruloplasmin (Cp), Total protein, Albumin, Calcium, Total bilirubin, Direct bilirubin, Uric acid, Creatinine, Total iron binding capacity (TIBC), Iron, Sulfate, Glutathione(GSH), Malondialdehyde (MDA), Cholesterol, Glucose, Selenium and Transferrin saturation(%). The study included (102) persons living outside city center represented into two minor groups, smokers and non smokers. The results showed that the level decreased significantly in rural smokers when compared with rural non smokers in: vitamin C, total bilirubin, direct bilirubin and MDA, and there was a significant increase in: vitamin A and Cp., but there wasn’t any significant changes in: total protein, albumin, uric acid, creatinine, TIBC, iron, sulfate, cholesterol, selenium, calcium, vitamin E, β-carotene, folic acid, GSH, glucose and transferrin saturation(%). Depending on the period of smoking, the results predicted that antioxidants and some other parameters (vitamin E, vitamin A, β-carotene, vitamin C, folic acid, total protein, albumin, calcium, total bilrubin, direct bilirubin and GSH) were decreased, while: Cp., uric acid, creatinine, TIBC, Iron, sulfate, MDA, cholesterol, glucose and selenium were increased with increased the period of smoking. ــــــ ــــــــــــ ـــــ ـــــــــــــــــــــــــــــــ ل يدسكأتلا بركلا ل نينخدم يف يحاوض لصوملا ةنيدم


INTRODUCTION
Smoking and related diseases account for approximately 440,000 American deaths every year.This number includes both the direct effects of smoking on the smoker, but also indirect effects of smoking, such as premature births and the effects of second hand smoke .The costs are enormous.Estimates of costs of smoking, both in terms of health care costs and related to lost productivity, are between 100-150 billion dollars each year (Zheng, 2003).
Cigarettes smoke contains carbon monoxide and free radicals which are generated because of high temperatures (up to 900 ºC) at the burning tip (Gillham et al.,2000).
The free radicals found in the tar phase are different to those in the gas phase with respect to stability and half-life.The tar phase contains long-lived and fairly stable radicals.It also contains transition metal ions, which can drive the formation of the reactive hydroxyl radical.The gas phase contains short-lived reactive radicals such as nitric oxide ( .NO).In the presence of oxygen, this is oxidized to nitrogen dioxide radicals, which can damage lung tissue (Gillham et al., 2000): . NO +1/2O 2 .NO 2 and contain reactive olefins and dienes.
A major exogenous source of free radicals is the cigarette smoke.The cigarette smoke is a complex mixture of approximately 5000 chemical compounds, including high concentrations of free radicals and other reactive oxygen species.These oxidants are contained in both the tar and gas phases of cigarette smoke.The gas-phase reactive oxidants are both inorganic and organic in nature, and include epoxides, peroxides, nitric oxide (NO), nitrogen dioxide, peroxynitrite (ONOO-), perinitrates, carbon monoxide, ammonia, dimethylnitrosamine, formaldehyde, hydrogen cyanide, acrolein and a myriad of other free radicals.Indeed, it has been reported that gas-phase cigarette smoke contains approximately one quadrillion radicals per puff.These represent an enormous oxidant load to body tissues (Zheng, 2003).
Damage to DNA appears to involve a complex of polyphenolic tar components with DNA, followed by production of the hydroxyl radical that nicks DNA (Zheng, 2003).
Health disease and lung cancer comprise the vast majority of deaths caused by smoking, followed by chronic bronchitis, strokes, peripheral vascular disease and other cancers.
The study included (102) persons (59 nonsmokers, 43 smokers) healthy subjects, with comparable age, all were males with minimal or no risk of exposure to a polluted atmosphere.They were farmers living in Al-Khather village (30 km east of Mosul city center).
For venipuncture, 10 ml sterile syringes equipped with (22G x 1.25) syringe needles were used and put in dry and clean plain tube.After coagulation, it was centrifuged at 4000 x g for 15 minute.Serum was transferred into plain tube equipped with tight-fitting caps by disposable tips, then stored at -20 ˚C (Liang et al., 1989).

RESULTS AND DISCUSSION
The results of smokers and non-smokers groups in suburbs group were listed in Table (2).The results showed that there was a significant decrease of suburbs smokers when compared with suburbs non-smokers in vitamin C(P=0.03).Similar results were published by other investigators (Riemersma et al., 1991;Sakai et al., 1998;Pincemail et al., 2000;Lykkesfeldt et al., 2000;Nuttall et al., 2002).In vivo vitamin C has been suggested to be a biomarker of oxidant stress, and the level of vitamin C was decreased under the oxidant stress of smoking (Turnlund et al., 2004).Low vitamin C levels found in smokers are considered to be a direct consequence of vitamin C utilization as an antioxidant due to the smoke-related pro-oxidant load (Nuttall et al., 2002).
Other biochemical parameters were also decreased significantly as: total bilirubin (P=0.01),direct bilirubin (P=0.005) and malondialdehyde (P=0.019).These results were in a good agreement with the previously reported data (Al-Timimi and Al-Khayat, 2001).
Vitamin E, β-carotene and folic acid were not significantly decreased.These results meet with those of (Riemersma et al., 1991;Durand et al., 1997;Hippeli et al., 2003).The decreased in vitamin A, found in smokers is considered to be a direct consequence of its utilization as an antioxidant due to the smoke-related pro-oxidant load (EBSCO, 2005) similar to other reported results (Kim et al., 2004).
Moreover, it has been observed that there was a significant increase in: ceruloplasmin (P=0.004) which were similar to other reported results (Al-Timimi and Al-Khayat, 2001;Sürmen-Gür et al., 2003).
The result of this study might suggest that the increase of Cp. levels in smokers is a part of the total antioxidant status protecting tissues from the effects of free radicals (Al-Timimi and Al-Khayat, 2001;Sürmen-Gür et al., 2003), or might merely reflect increase copper intake from the tar component of the cigarettes (Duthie et al., 1991).

Effects of the smoking period on the biochemical parameters in smoking suburb group.
Depending on the period of smoking, suburb smokers group was divided into three subgroups, which were listed in Table (3).
The results showed that vitamin E showed no changes with the increase of the period of smoking (Table 3).Notice that vitamin A significant decreased in (0-10 year) group and in (21year and over) group, but there were no significant differences between them.
β-carotene, vitamin C and folic acid were decreased significantly with the increase of the period of smoking.While no significant difference was observed in β-carotene.This result was in a good agreement with other reported results (Helen and Vijayammal, 1997).
Several studies have also examined the effect of vitamin E in cigarette smokers.Cigarette smoking stimulates the formation of highly reactive molecules.Serum levels of vitamin E (as well as vitamin C and β-carotene) react as antioxidant to these molecules.Therefore the level of vitamins were significantly lower in the smokers compared to the non-smokers, and antioxidant decreased too when the period of smoking increased (Hughes, 2000).Ceruloplasmin, on the other hand, increased with increasing the period of smoking and there was a significant difference within each one of the groups.
Total protein and albumin were significant difference in last groups (Table 3).Calcium, total bilrubin and direct bilirubin were not affected with increasing the period of smoking and this result was agreement with literature (Chan-Yeung et al., 1981;Vogt, 1999).The mechanism of action of smoking on these parameters is unclear (Vogt, 1999).Uric acid, creatinine, TIBC and Iron increased significantly with the increase of the period of smoking, while TIBC and iron showed non a significant differences.Similar results were published by others (Chan-Yeung et al., 1981;Ahmad and Al-Helaly, 2002;Mannino et al., 2004).
Sulfate was increased with increasing period of smoking, where a significant difference among the groups was observed.The increase in sulfate might be due to the different compounds present in the smoking, especially SO 2 ,H 2 S..etc.
Glutathion was decreased with the increase of the period of smoking and there was a significant difference among them.Nitric oxide in cigarettes might contribute to the GSH levels by directly reacting with glutathione and inhibiting antioxidants enzymes like glutathione reductase (Fechner et al., 2001).
Malondialdehyde wasn't affected, and this finding agrees with other observation that was found earlier (Helen and Vijayammal, 1997).
Cholesterol increased with increasing the period of smoking and showed significant difference in all groups as found in the literatures (Ahmad and Al-Helaly, 2002;Walker, 2003).The increase might be due to increase secretion of adrenaline hormone stimulated by the nicotine in cigarettes (Cryer et al., 1976).It is Known that adrenaline would increase the lipolysis and cholesterol level would be increased too (Burtis and Ashwood, 1999).
Glucose was increased with increasing the period of smoking and a significant difference among the groups was present.This finding agrees with the other observation found (Ko et al., 2001).It was reported that insulin concentration in smokers was decreased, and hence glucose would be increased (Burtis and Ashwood, 1999).
The results also showed that selenium was increased significantly with increasing the period of smoking and a significant difference among the groups was observed.
Transferrin Saturation (%) on the other hand was decreased significantly and a significant difference among them was observed.

RECOMMENDATIONS
1.Smoking might be a contributing factor in increasing lead absorption among exposed people.2.Avoidance of direct and indirect exposure to tobacco smoke is of primary importance not only for healthier lungs, but as a preventative measure for the other diseases: Cardiovascular disease, cancer, and diabetes, as well as the clinical importance of the present results suggesting that free radicals inhaled in cigarette smoke are highly toxic, and impaired oxidant-antioxidant balance is a risk factor in degenerative diseases.Therefore, may God help to prevent anything that affects the human body and lead to death: In the name of Allah Most Gracious Most Merciful.(The heifer) (195) {And spend of your substance in the cause of Allah and make not your own hands contribute to your destruction but do good; for Allah loveth those who do good} ), Albumin No. (0801), Calcium No.(2403),Total Bilirubin and Direct Bilirubin No.(0401),Iron No. (0502) ,Total Iron Binding Capacity No. (0512), Glucose No.(0903), Cholesterol No.(0603), were obtained from Syrbio kits, Syria.

Table 1 :
Methods used to determination of biochemical parameters.

Table 2 :
The biochemical parameters of suburbs non-smokers and smokers

Table 3 :
The biochemical parameters of suburb smokers at different period of smoking and control using Duncan test.