GERIATRIC SYSTEM BIOCHEMICAL CHANGE

**FREE RADICAL THEORY **TELOMERE

Department of Biochemistry Faculty of Medicine.Hasanuddin Univ. By Rosdiana Natzir

Free Radicals and Antioxidants

Free Radicals (FRs)

These are highly reactive chemical entities that have a single unpaired electron in their outer most orbit. Under certain conditions can be highly toxic to the cells. Generally unstable and try to become stable, either by accepting or donating an electron.

 Therefore if two FRs react, they neutralise each other . However, if the FRs react with stable molecules, there is generation of more free radicals.

This characteristic enables the FRs to participate in auto catalytic chain reactions,
Molecules with which they react are themselves converted to free radicals to propagate the chain of damages.

Reactive Oxygen Species (ROS): These are free radicals derived initially from oxygen. But as they do not contain unpaired electrons in their outermost orbit, they do not qualify as free radicals and so are referred to separately as ROS.

Eg.

- H2O2, HOCL, NO.

Free radicals are formed in side our body by both PHYSIOLOGICAL (Natural) and PATHOLOGICAL stimuli : 5

Physiological Stimuli that Form FRs

Normal respiration
O2 Superoxide, H2O2 Hydrogen Peroxide HOCL Hypochlorous acid NO Nitric Oxide

Transition metals present inside our body when are in free form behave as free radicals. Fe2+, Cu+

 Body cells Endothelium (NO3 Nitric Oxide, NO2 Nitrous Oxide), Macrophages (NO2) Neurones (ONOOH Peroxy nitrite).

Ageing Phagocytosis or biogenetics Oxidation of foods and endogenous compounds. Transportation of substances for energy production.
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Pathological Stimuli that Form FRs

Radiation Breaks the water inside our body: H2O =H+ + OH Metabolism of drugs CCl3 Transition Metals Cu+, Fe2+ Ultraviolet rays Emotional stress

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Actions of FRs
Mechanism of Action: They act on the cell membranes and membranes of different organelles of cells and cause cell injury and death by oxidative reactions. So FRs are also called OXIDANTS.

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Actions of FRs
FRs cause lipid peroxidation.
The PUFA of cell membrane are more vulnerable for this injury. By lipid peroxidation FR increases the permeability of cells, leading to calcium influx and altered PH of the cell.

FRs alter the enzyme and receptor proteins.

FRs cause cross-linking of proteins and fragmentation of protein strands, oxidation of amino acids like cystene, Methionine. These alterations in the enzymes and receptoors inside the cell lead to abnormal cell behaviour.

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Actions of FRs
FRs cause fracturing on the cell nucleus resulting in single strand DNA damage. This oxidative injury may be.
Lethal Leading to cell death and ultimately removed by phagocytosis. Sub lethal - which may result in
Increased cell permeability. Toxicity. Mutation of cells.

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FRs Induce Chain Reaction

During the process of oxidant damage resulting in tissue destruction & degeneration, some electrons may escape oxidation and become FRs This chain reaction may produce diseases like:
Carcinogenesis. Myocardial reperfusion injury. Shock related injury. Arteriosclerosis. Rheumatoid arthritis. Adult respiratory diseases. Diabetes. Obesity. Lipid abnormality. Etc.

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Antioxidants (AOs)
These are substances, derived from oxygen. In normal healthy state a balance is maintained between FRs & AOs The AO activity of serum is measured as % inhibition of lipid peroxidation in a standardised brain homogenate. Moreover we can as well supplement these from outside (in vitro Antioxidants).

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In Vivo Antioxidants: I.Enzymes

Name
SOD (Superoxide dismutase) CATALASE

Acts against
Super oxide

Present in
cytosol mitochondria. Blood, bone marrow, Mucus membrane Kidney; Liver.

H2 O2

GOP (Glutathion H2 O2, lipid peroxidase) peroxidation

membranes of lipids, Haemoglobin and erythrocytes

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II. Preventive AOs These are binding proteins. They keep the free ions of plasma in a binding form, so prevent oxidation injury. Eg.-Transferin for Fe, Ceruloplasmin for Cu
III. Scavenger AOs

Also called chain breaking enzymes, they break the catalytic chain propagated by FRs.
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In Vivo Antioxidants: - Source

I. In the form of Medicines:
Vitamin A, C & E,
Cystine, Glutathion, Melthionine, Bioflavines, Se, Zn.

II. Food sources:

Green & yellow vegetables Herbs: -Turmeric/kunyit, Garlic, Grape, Tea, Berries, Carrot, Spinach, Broccoli, Red Meat, Kidney, Liver & Lipoic Acid
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Oxidative Stress: Under normal conditions body maintains an equilibrium between its own FRs and Antioxidants. When this equilibrium breaks, a state called oxidation stress arises with in, due to FR formation or AO system.

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Example: AO Status in Normal Pregnancy:

There is increased need for AOs as there is increased production of FRs due to
Pregnancy being a stressful condition. Because of the rapidly growing foetus there is increased cellular activity.

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 Thus AO activity during normal pregnancy progressively increases as demonstrated by

Serum tocopherol. Activity of GOP(glutathione peroxidase) Serum ceruloplasmin & transferring level.

drugs that lead to FR formation must be avoided.

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AOs & Diabetic Embryopathy: Oxidative stress has been suggested to contribute to the increased risk of foetal malformations in poorly controlled diabetics.

lipid peroxidation in cell membranes in diabetic pregnancies Periods of maternal hyperglycaemia & hypoglycemia may cause marked changes in the availability of glucose to the foetus. conc. of lipids, notably the ketone bodies and branched chain amino acids in the maternal circulation contribute to altered nutrition for the embryo.
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 During later part of pregnancy

load of glucose in the mitochondria may accelerate the flow of electrons through respiratory chain including mitochondrial leakage of free radicals.
This leads to production of FR in embryonic tissues to cause congenital malformations.

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 Thus maintenance of normal concentration of metabolites of all nutrient class may be important for prevention of adverse foetal outcome. However, maintenance of blood glucose level at euglycemic level is always important for prevention of Diabetic embryopathy .

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AOs & Down Syndrome: Free radicals being the hallmark of aging, are greatly increased with maternal age. So FRs play a role in pathogenesis of Downs syndrome. Administration of AOs may help in preventing this disease.

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 Increased activity of free radicals promote maternal uterine vascular malformations.

FRs are promoters of maternal vasoconstriction.
O2 , H2 O2 & NO2 in combinations Inactivate the NO (a vasorelaxant) Causes PG synthatase activity. Produce peroxynitrate, a potent oxidant

Lipid peroxide in pre-eclamptic placenta is about 1.8 times higher in comparison to normal placenta. Severity of hypertension has been found to be inversely proportional to concentration of Vit. E.

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Carcinogenesis: Basics of cancer formation: A normal cell can undergo malignant transformation in presence of procarcinogens and carcinogens. Without immune system detection a normal cell is converted to malignant cell in stages >
Initiation Promotion Progression

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 Free radicals are formed from stimulants like Radiation - Xenobiotics - Inflammatory cells - Respiration etc, which act on cellular targets to cause oxidant DNA damage in form of mutagenesis.

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- which cause: - initiation of carcinogenesis by Activation of protooncogens. Inactivation or loss of tumour suppresser genes. Normal cell becomes initiated cell.

Procarcinogens are metabolically activated by FRs, which cause promotion and progression of these initiated cells to cancer

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FRs & AOs in Carcinogenesis: FRs Activation of

procarcinogens & Carcinogens Genetic Damage

Normal cell
Initiation

Repair by AOs

Initiated Cell
Promotion

FRs
- Tumour promoters - Spontaneous

Repair by AOs

Premalignant Cell
Repair by AOs Progression

FRs
- Dysregulation - Greater Cell autonomy - Reduced Growth Factor dependence

Malignant Cell
CLINICAL CANCER
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AOs in Cancer Prevention: DEFENCE:

- enzymes with antioxidant property cause first line of defence by: Protecting the lipids and enzymes against oxidation Creating a balance of AOs against FRs in the body PREVENTS :
Cell pathology. Metabolic disturbances. Changes in cell permeability. Eormation of toxic products.

Prevention of initiation of Carcinogenesis

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 INTERVENTION at promotion & progression stages:

Local deactivation of genotoxins responsible for further nuclear mutations Inactivation of tumour promoters eg.- activation of granulocytes. Simulate oxygen. Maintenance of proper function of gap junction communication. Maintenance of physical stability of membrane & also within cells.

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 Majority of epidemiological data suggest supplementation with antioxidants - Vit.- A, E, C; - beta carotene & selenium, decreases the incidence of various cancers.

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RECOMMENDED FOR CARCINOMA PREVENTION

Antioxidant RDA Recommende d Dose
12,500 IU

Possible Toxicity Level

Chronic intake of 125,000 IU >1,200 IU

Features Causing Req.

Smoking

Vitamin A

5000 IU

Vitamin E Vitamin C

10-20 IU 60 mg

200-800 IU 1000 mg

High PUFA intake, Smoking

Negligible / 1- Stress, 2Gms OCP,Smoking >200mg Aging, High PUFA intake, Smoking, Heavy metals
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Selenium

None

50-200 mg

 Various studies have suggested Conc. of Malon di-Aldehyde (MDA) is inversely proportional to fertility in case of males. In asthenospermic and oligoasthenospermic males there is increased serum concentration of MDA. Even in normospermic males if there is concentration of MDA there is reduced fertility. Addition of vitamin E causes decreased concentration of MDA and improves fertility.
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FRs & AOs in Infertility - Female

FRs cause
Short luteal phase . an arrest the cell growth (div) at 2, 4, 8 cell stages Hamper the regulation of corpus luteum.

Addition of AOs improves the results.

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CONCLUSION
Oxidant generation is a part of human life. Every O2 atom we breath in ,is converted to water inside our body by addition of four electrons sequentially. When four electrons are added three FRs- O2, H2 O2 & OH are formed along with water. So where there is life there are oxidants.
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CONCLUSION
But when produced in excess, they can cause any disease. So our concern should be to FRs in systemic circulation. However, careful use of AOs and newer and more accurate methods to measure oxidant generation in humans , will go a long way to find out the exact contribution of oxidants in disease processes and the role of AOs to prevent it.
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THANK YOU
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