INCREASED OXIDATIVE STRESS REALTED TO USE OF SMOKELESS

TOBACCO”MARAS POWDER”

Elif Köse¹ , Hakim Çelik², Özlem Yazıcıoğlu Moçin³

¹Ceylanpinar /Sanliurfa Tuberculosis Dispansery, Sanliurfa²Harran University School of Medicine Biochemistry Department, Sanliurfa³ TRMH Sureyyapasa Chest Diseases and Chest Surgery Training Hospital,

Istanbul.

FREE RADICALS • Having one or more paired

electrons,• Short half-life,• Unstable,• Low molecular weight,• Very active molecules

( H atom, O2 molcule, metal ions)

• Having an unstable structure, these molecules prefer to become stable.

• They have a property to interact with cellular elements because of instability in their structure.

SOURCES OF FREE RADICALS

A- Normal Biologic Processes: 1. Respiration with Oxygen,2. Catabolic and Anabolic

Reactions B- Conditions causing Oxidative

Stress• 1. Ischemia, hemorrhage,trauma,

radioactivity,• 2. Effects of Xenobiotics( Drug,

alcohol, tobacco),• 3. Oxidation of Catecholamines,• 4. Secretion from phagocytic

inflammatory cells,• 5. Long term metabolic diseases,• 6. Others: Hot shock, sunlight.C- Period of Ageing

FREE RADICALS

• The most important free radicals are derived from O2

• Inside cell: e- e -+2H+ e-+H + e-+H +

O2 O2-

H2O2 OH- H2 O• Oxygen is reduced by a chain of reactions requiring 4

electrons.• This supplies nedde energy for it. • 1-3% transform into free radicals. • 1,7 kg/year O2

- is formed in human body.

• According to O2 consumption, it may rise 10 times. .

FREE RADICALS

1. Radicals• Superoxide radical(O2)• Hydroxyl radical(OH)• Alcoxyl radical(LO)• Peroxycyl radical(LOO)• Nitric oxide(NO)• Nytrogen dioxide(NO2)2. Non-radicals• Hydrogenperoxide(H2O2)• Lipid Hydroperoxide(LOOH)• Hypocholorus(HOCl)3. Single Oxygen

ROS

DNALIPIDS

PROTEINS

GENETIC MUTATION

LOSS OF MEMBRANE

INACTIVATION OF ENZYMES

ANTIOXIDANTS

• Cells and tissues have a strong protection system inhibiting radical productions and reactions.

• Antioxidants are defined as substances that react rapidly with radicals preventing progress of autooxidation/peroxidation.

ANTIOXIDANTSA) Intracellular Antioxidants

Superoxide dismutaseCatalaseGlutathione peroxidaseCytochrome oxidase

B) Membrane AntioxidantsVitamin ECoenzyme-QB-Carotene

C) Extracellular AntioxidantsAlbuminUric acidAscorbic acidBilirubinTransferrin, Lactoferrin, Haptoglobulin, Seruloplasmin

OXIDATIVE STRESS• OXIDATIVE BALANCE

• In organism, production rate and clearance rate of free radicals is in an equilibrium.

• As long as oxidative balance is preserved, free radicals do not affect organism.

Free radical Antioxidant

OXIDATIVE STRESS

• If the production rate of free radicals exceeds the protective capacity of antioxidants, noxious effects of free radicals begin to appear.

Free radicalAntioxidant

THE DISEASES RELATED WITH OXIDATIVE STRESS and FREE

RADICAL INJURY-1• Cancer,

• Atherosclerosis, Ischemia-Reperfusion Injury,

• Injury of Radiation,

• Lung Diseases( Emphysema, Asbestosis, O2 toxicity),

• Diabetes Mellitus,

• Rheumatoid Arthritis and other autoimmune diseases,

• Disorders of brain(Alzheimer, Parkinson Dis),

• Ageing.

THE DISEASES RELATED WITH OXIDATIVE STRESS and FREE

RADICAL INJURY-2• Renal Disorders( Autoimmune Nephrosis),• Muscular Dystrophies(M sclerosis, Muscle

dystrophy),• Opthalmologic Disorders( Macular degeneration,

cataract),• Dermatologic Disorders( Burns, Contact Dermatitis),• Hepatologic Disorders,• Hematologis Disorders( Favism, Sickle cell anemia),• Gastrointestinal Disorders,• Nutritional Insufficiency( Kwashiorkor).

TOBACCO-OXIDATIVE STRESS

• Tobacco contains more than 4700 oxidant and free radicals.

• Free radicals are detected in both gas and tar phase of tobacco.

• Gas phase contains 1015 radicals in every puff ( alkyl and peroxil type)

• The carcinogenic and mutagenic agents in content of tobacco also exist in smokeless tobacco products in different amount.

TOBACCO-OXIDATIVE STRESS

The lungs have strong and diffuse antioxidant system in intracellular and extracellular level:

• Catalase• Superoxide dismutase,• Glutathione peroxidase,• Ascorbic acid,• Tocoferol,• Carotene,• Albumin,• Uric acid.

MARAS POWDER “CRAZY TOBACCO”

• The types and use of smokeless tobacco( chewable snuff) differ according to countries.

• The most frequent use of smokeless tobacco is seen in West Europe, America, South Africa, south part of Arabia.

• Maras powder is ferquently used in K.Maraş in Southeast region, Gaziantep and Adıyaman.

MARAS POWDER “CRAZY TOBACCO”

• It is obtained from Nicotiana rustica Linn plant.

• The alkoloid composition of it is similiar to Nicotiana tabacum L, culture tobacco for cigarettes.

• The nicotine content of Maras powder is 6-10 times higher than cigarette.

MARAS POWDER “CRAZY TOBACCO”

• Dried Nicotiana rustica L is prepared by mixing the ash of oak, walnut and vine stem in ratio of ½-1/3.

• The admixed ash makes the medium alkaline and facilitates the absorption from mouth mucosa.

• This feature increases physical dependence for Maras powder.

MARAS POWDER “CRAZY TOBACCO”

• A sweet spoon amount of mixture is placed between the lower lip and chin.

• After being kept for 5-10 minutes within mouth, tobacco and saliva mixture is expectorated.

• This procedure is repeated 10-20 times a day and some people even sleep with the powder in their mouths.

MARAS POWDER “CRAZY TOBACCO”

It has been detected that Maras Powder, which has been used as an alternative for tobacco, has adverse

effects for human health: Biochemical,

Immunological,Hematologic parameters,

Worsening in lung functions like in case of tobacco,Oral cancer risk,

Genotoxic effects,Cardiovascular Diseases.

AIM

• To investigate effects of “Maras Powder” use on serum total antioxidant and oxidant levels

• To compare “Maras Powder” effects with that of tobacco.

MATERIAL-METHOD-1

• The study included people living in Ceylanpinar district of Sanliurfa city in September 2006.

• People without any respiratory symptoms, with no

history of medicine or antioxidant use and without comorbid conditions like metabolic, endocrine or malignant diseases were included in the groups.

MATERIAL-METHOD-2

• Group 1(Control):24 males between 20-51 year of age ( mean value of age 30,08 ±6,69), no history of tobacco use.

• Group 2( Smokers): 27 males between 25-75 year of age ( mean value of age 37,7 ±10,21), with at least 10 pack year history of smoking

• Group 3(Maras powder users): 21 males between 20-76 year of age (mean value of age 46,81 ±16,34), with at least 5 years use of one pack/day (16 gr of Maras powder)

MATERIAL-METHOD-3

• 10 ml of venous blood was taken from each subject after a night of fasting and serum part was separated by centrifugating 10 minutes at 3000 turns.

• The sera of patients were labeled and kept at -20° C untill the day of analysis.

MATERIAL-METHOD-4

MATERIAL-METHOD-5

RESULTSControl Smokers Maras

powderP

Mean age (yr)

30,08 ±6,69 37,7 ±10,21 46,81 ±16,34 p<0,05

Duration(yr) - 20,78 ± 11,08 22,38 ±18,58 p>0,05

Number of smoked cigarette

- 22,96±6,09 -

Frequency of use

- - 17,95 ±9,46

Duration of Maras powder use- Total Oxidant Stress

0

2

4

6

8

10

12

0 10 20 30 40 50 60 70

duration of Maras powder use

Tota

l oxi

dant

leve

l

oxidant stree

• There was no relation between duration of Maras powder use and TOS (p>0.05).

Duration of Maras powder use-Total Antioxidant Capacity(TAC)

• There was a negative correlation between duration of Maras powder use and TAC

• (p<0.05, r=-0.435).

0

0,5

1

1,5

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2,5

0 10 20 30 40 50 60 70

duration of maraş powder use

Tota

l ant

ioxid

ant c

apac

ity

TAO

TOTAL OXIDANT LEVEL(TOL)

• According to TOL, there was a significant difference between smokers group (mean value 11,21 ± 3,54) and control group(mean value 8,83±1,28 ) (p<0.05).

• According to TOL, there was no significant difference between smokers and Maras powder users group(mean value 9,73± 2,68 ) (p>0.05).

0

2

4

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12

kontrol sigara maraş

Total Oxıdant level

TOTAL ANTIOXIDANT CAPACITY(TAC)

• According to TAC, there was a significant difference between Maras powder users(mean value 1,35 ± 0,18 ) and control group(mean value1,57±0,17 ), between Maras powder users and smokers group (mean value 1,48 ± 0,11 ) (p<0.05).

1,2

1,3

1,4

1,5

1,6

kontrol sigara maraş

Total Antıoxıdant capacıty

OXIDATIVE STRESS INDEX(OI) OXIDATIVE STRESS

INDEX(OI)= TOL/TAC • Control group: 5,67 ± 0,99

• Smokers group:7,62 ± 2,61

• Maras powder users: 7,44 ± 2,84

There was a significant difference between Maras powder group, smokers group and control group (p<0.05).

There was no significant difference between Maras powder group and smokers group(p>0.05).

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kontrol sigara maraş

oksidatif stres indeks

oksidatif stres indeks

CONCLUSION

• The use of Maras powder and smoking both increase total oxidant level.

• In tobacco users, total antioxidant capacity declines because of increased oxidant load and this decrease is much more significant in Maras powder users.

CONCLUSION

• The use of Maras powder leads to oxidative stress as much as smoking in organism.

• It is concluded that preference of Maras powder as a way of quiting smoking or decreasing dependence for tobacco is not a correct way for smokers and on the contrary it has noxious effects like tobacco.