1. INTRODUCTION:

Use of tobacco has become topic of attention for researchers and public health policy makers worldwide. Tobacco is consumed in many ways such as smoking, chewing and snuffing in powder form [1]. Tobacco use is familiar throughout the global due to its widespread marketing, addiction potential of nicotine, absence or weakness of warnings and legislation concerning tobacco use and low price [2]. Tobacco is the leading cause of morbidity and mortality among the young and the elderly [3]. According to report of World Health Organization (WHO) tobacco is considered as the single greatest cause of preventable death globally [4]. Direct tobacco use is estimated to cause five million deaths a year globally, while indirect exposure leads to an additional 600, 000 deaths [5]. Many of recent landmarks in scientific research have shown that in human beings, Oxidative stress is an important factor causing metabolic and physiological alterations and various diseases in the body [6]. Tobacco and their products are rich by pro-oxidants that are proven to induce oxidative stress. Smoking of tobacco is done in various forms like cigar, cigarette, beedi, hukka, pipe…etc [7]. Cigarette production and consumption have seen dramatic growth in recent decades [8]. Cigarette smoke contains more than 10¹⁴ low-molecular weight carbon and oxygen centered radicals including peroxyradicals, nitrogen radicals and other oxygen-derived species [9]. The toxic products or free radicals present in cigarette smoke are thought to activate inflammatory immune responses, which may play an important role in smoking-related oxidative tissue damage that have a significant role in the pathogenesis of diseases. Smoking is considered to cause heart disease, cancer, stroke and also have close relationship with gastric ulcer, periodontal disease and metabolic syndrome. [10,11]. Smoking has significant detrimental effect on various systems of body especially on cardiovascular system. It is also a factor associated with death premature [12,13]. The aim of our work is based on the realization of some analyses of lipid profile, biochemical and hematological biomarkers and the estimation of blood pressure on Algerian men for study the effect of tobacco smoking.

2. MATERIALS AND METHODS:

Ethical approval (Appendix) was sought and approved by the Ethical Committee of the Department of Cellular and Molecular Biology, Faculty of natural sciences and life, University of El-Oued. This study was conducted on 22 volunteers man adult (mean aged 30.97±1.33 years), were divided into two groups; habitual smokers and non-chewers (11 subjects) when the mean consumption 19.91 ±2.28 cigarettes per day and mean duration of the consumption 13.64±2.88 years, and 11 healthy volunteers served as controls, were joined in this study. All the volunteers in this study live in the El-Oued area located in the southeast of Algeria.

Inclusion Criteria:

The populations of our study included the habitual smokers for more than 5 years and non-tobacco consumers.

Exclusion Criteria:

To eliminate the factors which might affect lipid profile and biochemical parameters, we excluded all diabetic, high blood pressure and other chronic diseases subjects from chewers and healthy controls groups.

Laboratory Investigations:

Fasting blood samples were collected and transferred into EDTA tubes for hematological analysis and non-heparinized tubes for lipid profile and biochemical analysis. The serum was obtained by centrifugation of the blood at 3000 × rpm for 5 min and stored at -20°C until use. The samples were collected between 08:00 hours and 10:00. The determination of hematological parameters performed using fully Auto Blood Cell Counter. The following serum biochemical and lipid parameters were measured using commercial kits (Biomaghreb, Tunisia): blood sugar, serum urea, serum creatinine, aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (TC) and High-Density Lipoprotein-cholesterol (HDL-c) levels. Low-Density Lipoprotein-cholesterol (LDL- c), Very Low-Density Lipoprotein-cholesterol (VLDL- c) were calculated by using Freidewald et al (1972) [14] formula:

VLDL-c = TG/5; LDL-c = TC - (HDL-c + TG/5) and

Atherogenic Index (AI) = TC/ HDL-c according to Gomina et al (2013) [15].

Statistical methodology:

The statistical evaluation is carried out by the student T test using Minitab software (version 13.31). The reported data are the means of measurements and their standard error of mean (SEM) values.

3. RESULTS:

The blood pressure results presented in table 1, demonstrated that there was a significant increase (P <0.001) in the levels of SBP in smokers compared to controls.

Table 1. Mean of systolic blood pressures (SBP) and diastolic blood pressures (DBP) in tobacco smokers and controls groups

Blood pressure	Controls	Smokers	P-value
SBP (mmHg)	11.79±0.283	12.955±0.34	0.000
DBP (mmHg)	7.73±0.295	8.1±0.252	0.081

Values are mean ± SEM (n=11). *** p<0.001: significantly different compared to the control group.

Table 2 presented the different means of lipid profile, when the results showed a significant increase in TG, VLDL-c and LDL-c levels (p<0.01), additionally to TC level (p<0.05) in smokers group when we compared to the controls one. However, there are a significant decrease (p<0.001) in HDL-c level.

Table 2. Mean lipid level in tobacco smokers and controls groups

Lipid profile	Controls	Smokers	*P-value*
Serum Total Cholesterol (g/l)	1.635± 0.081	1.822±0.082	0.041
Serum Triglycerides (g/l)	0.758±0.072	1.107±0.106	0.007
Serum HDL-c (g/l)	0.526±0.019	0.464±0.014	0.000
Serum LDL-c (g/l)	0.969±0.070	1.220±0.078	0.003
Serum VLDL-c (g/l)	0.154±0.016	0.224±0.02	0.005

Values are mean ± SEM (n=11). * p<0.05, ** p<0.01and *** p<0.001: significantly different compared to the control group.

The Atherogenic index (AI) of smokers were found to be statistically significant elevated (p<0.001) compared to controls as demonstrated in figure 01.

Figure 1. Variation of the mean Atherogenic index (AI) in tobacco smokers and controls groups.

Values are mean ± SEM (n=11). *** p<0.001: significantly different compared to the control group.

The Biochemical parameters were non-significant changed (p >0.05) which including the blood sugar, AST, serum urea and creatinine levels in smokers compared to controls (table3).

Table 3. Mean serum biochemical parameters in tobacco smokers and controls groups

Biochemical parameters	Controls	Smokers	*P-value*
Blood Glucose (g/l)	0.911±0.0299	0.9075±0.0224	0.435
Serum Urea(g/l)	0.1945±0.0099	0.2145±0.0131	0.231
Serum Creatinine (mg/l)	9.385±0.789	8.718±0.335	0.098
Serum AST(UI)	23.64±1.61	23.00±1.38	0.365

Values are mean ± SEM (n=11).

Concerning hematological parameters (table 4), Our results demonstrated that there was a significant decrease in RBC (p<0.01) and PLT (p<0.001) levels while a significant increase in MCV (p<0.05) level in smokers group compared to the controls. Concerning leukocyte line, the results showed a significant raise of WBC (p<0.05) and Lym (p<0.001) levels in smokers group compered to controls..

Table 4. Erythrocyte and Leukocyte line in tobacco chewers and controls groups

Parameters	Controls	Smokers	P- value
Red blood cell (10⁶/µl)	5.255±0.153	4.878±0.125	0.008
Hemoglobin (g/dl)	14.440±0.316	14.156±0.293	0.685
Hematocrit (%)	44.86±1.28	42.958±0.947	0.325
Mean Corpuscular Volume (FL)	85.68±2.12	88.70±1.17	0.024
Platelet (10³/µl)	203.6±11.7	174.22±5.11	0.000
White blood cell (10³/µl)	6.091±0.294	6.747±0.285	0.031
Lymphocytes (10³/µl)	2.139±0.181	2.6211±0.0955	0.000

Values are mean ± SEM (n=11). *p<0.05, **p<0.01 and ***p<0.001: significantly different compared to the control group.

4. DISCUSSION:

The aim of this study to assessed the extent effect of smoking on blood pressure, lipid profile, biochemical and hematological characteristics of blood among Algerian men. Tobacco smoke is the main preventable lifestyle risk factor causing adverse health effects [16]. It has long been established, that tobacco contains nicotine, and that it has a considerable influence on the increasing levels of lipids in the blood [17]. In our study, it has revealed that tobacco smokers have a higher increased triglyceride, total cholesterol, VLDL-c, LDL-c levels and decreased HDL-c level as compared to controls. The nicotine is the active ingredient in tobacco [18], as the major component in smoke tobacco [19]. When cigarette smoke is inhaled, nicotine moves quickly to the lungs, arterial blood, and the brain in only 15 to 20 seconds, where it exerts its addiction-related effects. Rapidity of delivery to the brain is thought to be an important factor in the abuse liability of inhaled nicotine compared to other routes of nicotine administration [20]. The nicotine stimulates sympathetic adrenal system leading to increase the secretion of catecholamines [21]. This condition is followed by synthesis and secretion of hepatic free fatty acids, TG and VLDL-c into the blood stream. Additionally, tobacco smoking also induces the reduction of lipolysis enzymes, such as lipoprotein lipase (LPL) activity, in adipose and muscular tissue, thus reducing TG hydrolysis and clearance [22]. Several studies reported that Cigarette smoking is known to be associated with raised plasma Homocysteine level that may lead to decrease HDL-c level by several mechanisms. Which may be cause the oxidative modification of LDL-Cholesterol and decreases HDL-Cholesterol, or inhibit the Apo A-1 protein expression and decreases HDL-c in blood [23]. So, the increase in LDL-c and decrease in HDL-c concentrations. Also HDL-c concentration was inversely related to VLDL-c concentration in serum [24]. The increase in these lipid parameters resulting from smoking increases the likelihood of developing cardiovascular disease due to its direct relationship with the function of the heart and blood [25].

The mean values of Atherogenic indice (AI) was statistically significantly very highly increase in smokers as compared to controls group, Similar findings were observed in Ayu et al (2018), Therefore, the augment of total cholesterol and the decrease of HDL-c make the increase of atherogenic indice, which may be predicting cardiovascular risk in various clinical settings [26]. Previous studies confirm that blood pressure is raised in tobacco consuming, due to the sympathico adrenal-activating properties of nicotine and their acute effects in system cardiovascular [27]. Thus smoking promotes Coronary Heart Disease and atherosclerosis by lowering the anti-atherogenic factor HDL-c and increasing the potentially Atherogenic lipoproteins LDL-c which further may lead to vascular endothelium damage.

Cigarette smoking regulates both innate and adaptive immunity and causes numerous diseases, including cardiovascular, respiratory, and autoimmune diseases, allergies and cancers. The complex roles for smoking in immunoregulation are likely due to molecular and functional diversities of cigarette smoke components, including carbon monoxide and nicotine [28]. Nicotine present in tobacco may influence the suprarenal glands to secrete more catecholamine which may affect leukocytosis causing damage to tissue and inflammation [29]. Meanwhile, smoking has been implicated in the production of many immune or inflammatory mediators, including both pro-inflammatory and anti-inflammatory cytokines. One of the mechanisms of the xenobiotic like heavy metal, pesticide, cigarette’s effects is the occurrence of acute inflammation, which in turn increase the harm of these substances on various biological systems [30]. When, adaptive immune cells affected by smoking mainly include T helper cells (Th1/Th2/Th17), CD4+CD25+ regulatory T cells, CD8+ T cells, B cells and memory T/B lymphocytes while innate immune cells impacted by smoking are mostly DCs, macrophages and NK cells [31].

The decrease of red blood cells (RBC) level confirmed by study of Sharif et al (2014) who show that the administration of nicotine causes the diminished proliferation of red blood cells and that the low erythrocytes count may lead to a number of physiological disorders that may affect the efficiency of various enzymes [32]. A decrease in RBC necessarily leads to anemia, which causes a deficiency in the lack of oxygen in the body [33]. Additionally, the elevation of MCV level in our study is assisted by Acik et al (2020) [34]. Possible explanation for this situation, might be due to paucity of folic acid or vitamin B12 or thyroid problems in smoker’s subjects [35]. Singh and Singh (2019) suggest that the nicotine may be decreased the PLT count according to their experience on mice, but they say that the literature reports on the effects of nicotine on PLT count seem to be controversial, which show that the nicotine caused platelet and leukocyte activation [36].

5. CONCLUSION:

We conclude that the habitual consumption of cigarette leads to a remarkable alteration in the biomarkers which including a hyperlipidemia status that contribute to altered the blood pressure. Thus, the elevation of cardiovascular risk rate for tobacco smokers. Also, cigarette smoke may induce change in blood constituents and appearance of inflammation status in the body.

6. ACKNOWLEDGEMENTS:

We would like to thank the Faculty of Sciences of Nature and Life, University of El Oued, Algeria for the permission to utilize the facilities to make this work.

7. AUTHOR CONTRIBUTIONS:

SD: Conception, analysis, drafting, approval of the final version.

SC, OA, IB, IYG; Conception, design, data acquisition, approval of the final version.

8. FUNDING:

This research was supported by El-Oued University financially.

9. CONFLICT OF INTEREST:

Authors declare no conflict of interest.

Informed consent:

Written and Oral informed consent was obtained from all individual participants included in the study. Additional informed consent was obtained from all individual participants for whom identifying information is included in this manuscript.

Ethical approval for human:

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research (ethical approval code: (81 EC/CMB/FNSL/EU2020).

Data and materials availability:

All data associated with this study are present in the paper.

10. REFERENCES:

1. Patil SS, Patil SN, Somade PM. Comparative study of Pulmonary Function tests in Tobacco chewers and non-tobacco chewers. Indian Journal of Basic and Applied Medical Research; 2018; 8 (1): 208 – 214.

2. Bakdash A. Shammah (Smokeless Tobacco) and Public Health. Asian Pac J Cancer Prev, 2017; 18 (5): 1183-1190. Doi:10.22034/APJCP.2017.18.5.1183

3. Ahmed QR, Gupta N, Goyal S, Ansari S J. Comparative study on lipid profile in tobacco chewers and nontobacco chewers. National Journal of Physiology, Pharmacy and Pharmacology, 2015; 5 (2): 142 – 144.

4. Radha A R, Anjana S B. Comparison of Lipid Profile Levels in Tobacco Chewers with and without Leukoplakia–A Hospital Based Study. Annals of Medical and Health Sciences Research, 2017; 7: 341-345.

5. Kwamena S D, Bright O A. Understanding tobacco use and socioeconomic inequalities among men in Ghana, and Lesotho. Dickson and Ahinkorah Archives of Public Health, 2017; 75 (30): 1-8.

6. Derouiche S, Degachi O, Gharbi K. Phytochemistry analysis and modulatory activity of Portulacae oleracea and Aquilaria malaccensis extracts against High-fructose and high-fat diet induced immune cells alteration and heart lipid peroxidation in Rats. International Research Journal of Biological Sciences. 2019; 8(4): 6-11.

7. Amit D S, Shylaja T V, Asmabi M, Zahoorunissa D. Study of Lipid Profile among Healthy Smokers and Non Smokers. International Journal of Biotechnology and Biochemistry, 2017; 13 (1): 87-94.

8. Zafeiridou M, Hopkinson N S, Voulvoulis N. Cigarette smoking an assessment of tobacco’s global environmental footprint across its entire supply chain and policy strategies to reduce it. Geneva World Health Organization, 2018; 1-48.

9. Padmavathi P, Raghu PS, Reddy VD, Bulle S, Marthadu SB, Maturu P, Varadacharyulu NC. Chronic cigarette smoking-induced oxidative/nitrosative stress in human erythrocytes and platelets. Mol Cell Toxicol, 2018; 14:27-34. Doi: 10.1007/s13273-018-0004-6.

10. Derouiche S, Abbas K, Djermoune M. Changes in metabolism of Zinc and carbohydrate and testis oxidative stress of diabetic rats fed zinc-over dose diet. Int J Biol Med Res. 2017; 8(3):6041-6045.

11. Joshi N, Shah, Mehta HB, Gokhle PA. Comparative study of lipid profile on healthy smoker and non smokers. International Journal of Medical Science and Public Health, 2013; 2:622-626.

12. Samir D, Dalal D, Noura A. Effect of routine iron supplementation on copper level and oxidative stress status in pregnant women. Asian Pac J Reprod 2020; 9(2): 64-69.

13. Croizet J, Perriot F, Merson B, Aublet C. Sevrage tabagique des fumeurs agés Etude rétrospective chez 181 fumeurs agés pris en charge en centre de tabacologie. Revue des Maladies Respiratoires, 2016 ; 33 :241-247.

14. Friedewald WT, Levy RI, Fredrickson DS. Estimation of concentration of low-density lipoprotein cholesterol in plasma without use of ultracentrifuge. Clin Chem, 1972; 18: 499-502.

15. Gomina M, Ngobi GY, Akpona SA. Profil des lipides seriques des sujets adultes beninois consommateurs habituels de tabac. European Scientific Journal, 2013; 9(30): 125– 140.

16. Bauer M, Fink B, Thürmann L, Eszlinger M, Herberth G, Lehmann I. Tobacco smoking differently influences cell types of the innate and adaptive immune system—indications from CpG site methylation. Clinical Epigenetics, 2016; 8(83). Doi: 10.1186/s13148-016-0249-7

17. Srinivasa RCH, Emmanuel SY. The Effect of Chronic Tobacco Smoking and Chewing on the Lipid Profile. Journal of Clinical and Diagnostic Research, 2013; 7(1): 31-34.

18. Haragopal R, Aruna BMK. Effects of Tobacco Chewing on Serum Lipid Profile in South Indian Population. International Journal of Scientific Study, 2016; 4(4):9-11.

19. Salman S, Azmat R, Sangeeta S, Deepak P, Shadab K. Assessment of Cardiovascular Risks of Tobacco Chewers by Comparing it with Normal Human Beings. National Journal of Physiology, Pharmacy and Pharmacology, 2014; 4(1): 76 – 79.

20. Judith J. Prochaska and Neal L. Benowitz. Current advances in research in treatment and recovery: Nicotine addiction. Science advances, 2019; 5: 9763.

21. Elhashimi EH, Gabra HM. , Abdalla ZA. , Ali AE. . Effect of Cigarette Smoking on Lipid Profile In Male at Collage of Police and Low Khartoum, Sudan. Asian Journal of Biomedical and Pharmaceutical Sciences, 2013; 3(26):28-31.

22. Zulkefli B. Sanip, BSc, Muhammad Z.B. Suhaimi, BSc, Che N. Man, MSc, Aida H.B.G. Rasool, PhD and Harmy B.M. Yusoff, MMed. Relationship between hair nicotine levels with blood pressure, body composition, lipid profile and leptin among healthy male smokers in Kelantan. Journal of Taibah University Medical Sciences, 2016; 11(1): 50-55. Doi: 10.1016/j.jtumed.2015.11.007

23. Deepa S. Effect of Cigarette Smoking on Serum Lipid Profile in Male Population of Udaipur. Biochem Anal Biochem, 2016; 5(3), 1000283. DOI: 10.4172/2161-1009.1000283

24. Ramachandran M, Chinnasamy R, Ponniah T. Lipid and lipoprotein profiles among middleaged male smokers: a study from southern India. Tobacco Induced Diseases,2010; 8(11): 1-5.

25. Derouiche S, Kechrid Z. Influence of calcium supplements on zinc status, carbohydrate metabolism and the liver activity of detoxifying glutathione enzymatic system in alloxan induced diabetic rats. J Exp Biol Agr Sci. 2013;1(6): 424-429.

26. Ayu A, Ameh E A. Atherogenic indices and smoking habits in cigarette smokers. Environmental Disease, 2018; 3(2): 38-44.

27. Kjell A. Smokeless Tobacco and Cardiovascular Disease. Progress in Cardiovascular Diseases, 2003; 45 (5): 383-394.

28. Feifei Q, Ping F, Golay DN, Huazhen L, Chun-Ling L, Wanlin Y, Zhenhua D. Effects of Cigarette Smoking on Transplant Survival: Extending or Shortening It?. Frontiers in Immunology, 2017; 8. Doi:10.3389/fimmu.2017.00127

29. Dr Devendra Kumar, DR. B.K.Binawara, Mr. Poona Ram Beniwal, Mrs Pooja Sharma. Effect of Chewing Tobacco on Hematological Parameters in Bikaner City Population. JMSCR, 2017; 5(2): 17721-17727. Doi: 10.18535/jmscr/v5i2.87.

30. Derouiche S, Rezzag-mohcen OS, Serouti A. Triazinone herbicide metribuzin induced acute liver injury: A study of animal model. J Acute Dis 2018; 7(4): 152-157. doi:10.4103/2221-6189.241016.

31. Feifei Q, Chun-Ling L, Huazhen L, Yu-Qun Z, Shaozhen H, Song H, Xiaoping L, Zhenhua D. Impacts of cigarette smoking on immune responsiveness: Up and down or upside down? Oncotarget, 2017; 8(1): 268-284.

32. Sharif S, Farasat T, Nargis F, Farooq A, Naz . Effect of Nicotine on Hematology, Lipid Profile and Liver Enzymes in Adult Male Mice (Mus Musculus). Advances in Animal and Veterinary Sciences, 2014; 2 (4): 222 – 225. Doi: 10.14737/journal.aavs/ 2014/2.4. 222.225

33. Atoussi N, Guediri S, Derouiche S. Changes in Haematological, Biochemical and Serum Electrolytes Markers in Women Breast Cancer Patients. Scholars Journal of Research in Agriculture and Biology. 2018, 3(2): 73-177.

34. Acik DY, Suyani E, Aygun B, Bankir M. The Effect of Smoking on Hematological Parameters. Ulutas Med J2020; 6(1):9-14. Doi :10.5455/umj.20200209092535.

35. Bashir BA, Gibreel MO, Abdalatif HM, Mohamed MA, Ahmed EA, Mohamed MS and Hamid KA. Impact of Tobacco Cigarette Smoking on Hematologic parameters among male Subjects in Port Sudan Ahlia College, Sudan. Scholars Journal of Applied Medical Sciences, 2016; 4(4A):1124-1128.

36. Romsha S, Kalpana S. Protective activity of Guduchi and green tea extract on nicotine-induced toxicity in mice. The Pharma Innovation Journal, 2019; 8(1): 650-656.

Received on 07.09.2020 Modified on 18.09.2020

Asian J. Research Chem. 2020; 13(6):489-493.

DOI: 10.5958/0974-4150.2020.00086.3