SYSTEM GERIATRIC

Free Radicals and

Antioxidants

Dep.of Biochemistry
Faculty of Medicine.Hasanuddin Univ.

TOPICS

THE FREE RADICAL THEORY.

MITOCHONDRIA AND AGING.
THE GLYCATION THEORY.
PROTEINS DAMAGE AND MAINTENANCE IN AGING.
DNA DAMAGE AND DNA REPAIR.
TELOMERES THEORY.

Introduction

Therewasanerawhenpeoplewereworriedabout
thediseaseprocessesanditstreatment.
Butofflate,tremendouseffortsarebeingmadetofindout
thewaysof prevention of important diseases.
The important research studies are rightly to be repeated
with other populations, but nevertheless give an
opportunitytoconsidertheintegratedactionsoftheknown
reducing (Antioxidant) systems.
Inadequacy of certain internally produced reducing
systems has been postulated as being at the core of
some disease processes.

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It is believed that life has originated from

basic chemicals.
By free radical reaction, largely initialled by
ionising radiation from sun.
Paradoxically the same reactions creating
life may
also
be responsible for many
diseases, ageing and death.

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Free Radicals
Thesearehighly reactive chemical entitiesthat
havea single unpaired electron intheirouter most
orbit.
Undercertainconditionscanbehighly toxic to the
cells.
Generallyunstableandtrytobecomestable,either
byacceptingordonatinganelectron.

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 ThereforeiftwoFRsreact,theyneutraliseeachother
.
However,iftheFRsreactwithstablemolecules,there
isgenerationofmorefreeradicals.
ThischaracteristicenablestheFRstoparticipatein
autocatalyticchainreactions,
Moleculeswithwhichtheyreactarethemselvesconvertedto
freeradicalstopropagatethechainofdamages.

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ReactiveOxygenSpecies(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
separatelyasROS.
Eg.

- H2O2, HOCL, NO.

Freeradicalsareformedinsideourbodybyboth
PHYSIOLOGICAL(Natural)andPATHOLOGICAL
stimuli:-
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TheyaretwoconditionsofstimulitoformFRs:
1.Physiologicalstimuli.
2.Pathologicalstimuli.

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PhysiologicalStimulithatFormFRs
Normalrespiration

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

Transitionmetalspresentinsideourbodywhenarein
free form behaveasfreeradicals.Fe2+, Cu+

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 Bodycells Endothelium (NO3 Nitric Oxide, NO2 Nitrous Oxide),

Macrophages (NO2)
Neurones (ONOOH Peroxy nitrite).

Ageing
Phagocytosisorbiogenetics
Oxidationoffoodsandendogenouscompounds.

Transportationofsubstancesforenergyproduction.
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PathologicalStimulithatFormFRs
RadiationBreaksthewaterinsideourbody:
H2O=H++OH MetabolismofdrugsCCl3
TransitionMetalsCu+,Fe2+
Ultravioletrays
Emotionalstress

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ActionsofFRs
MechanismofAction:
Theyactonthecellmembranesand
membranesofdifferentorganellesofcellsand
causecellinjuryanddeathbyoxidative
reactions.
SoFRsarealsocalledOXIDANTS.

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ActionsofFRs
FRscauselipidperoxidation.
ThePUFAofcellmembranearemorevulnerable
forthisinjury.BylipidperoxidationFRincreases
thepermeabilityofcells,leadingtocalciuminflux
andalteredPHofthecell.

FRsaltertheenzymeandreceptorproteins.
FRscausecross-linking of proteins and
fragmentationofproteinstrands,oxidationof
aminoacidslikecysteine, Methionine.
Thesealterationsintheenzymesandreceptors
insidethecellleadtoabnormalcellbehaviour.
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ActionsofFRs
FRscausefracturingonthecellnucleus
resultinginsingle strand DNA damage.
Thisoxidativeinjurymaybe.
LethalLeadingtocelldeathandultimately
removedbyphagocytosis.
Sublethal-whichmayresultin
Increasedcellpermeability.
Toxicity.
Mutationofcells.

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FRsInduceChainReaction
Duringtheprocessofoxidantdamageresultingin
tissuedestruction&degeneration,someelectrons
mayescapeoxidationandbecomeFRs
Thischainreactionmayproducediseaseslike:

Carcinogenesis.
Myocardialreperfusioninjury.
Shockrelatedinjury.
Arteriosclerosis.
Rheumatoidarthritis.
Adultrespiratorydiseases.
Diabetes.
Obesity.
Lipidabnormality.Etc.
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Antioxidants (AOs)
Thesearesubstances,whichprotectusagainst
potentiallyharmfulfreeradicalsderivedfromoxygen.
TocombattheinjuriouseffectsofFRsourbodyhas
itsownsystemofin vivo Antioxidants.
Innormalhealthystate a balance ismaintained
betweenFRs&AOs
TheAOactivityofserumismeasuredas-%
inhibitionoflipidperoxidationinastandardisedbrain
homogenate.
Moreoverwecanaswellsupplementthesefrom
outside (in vitro Antioxidants).

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In Vivo Antioxidants: I.Enzymes

Name

Actsagainst

Presentin

SOD(Superoxide
dismutase)

Superoxide

cytosolmitochondria.

CATALASE

H2O2

Blood,bonemarrow,
Mucusmembrane
Kidney;Liver.

GOP (Glutathion H2O2,lipid

peroxidase)
peroxidation

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membranesoflipids,
Haemoglobinand
erythrocytes

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In Vivo Antioxidants: II.PreventiveAOs

Thesearebindingproteins.Theykeepthe
freeionsofplasmainabindingform,so
preventoxidationinjury.Eg.-Transferinfor
Fe,CeruloplasminforCu
III.ScavengerAOs

Also called chain breaking enzymes, they

breakthecatalyticchainpropagatedbyFRs.
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InVivoAntioxidants:-Source
I. IntheformofMedicines:
VitaminA,C&E,
Cystine,Glutathion,Melthionine,
Bioflavines,Se,Zn.

II.Foodsources:
Green&yellowvegetables
Herbs: -Turmeric/kunyit, Garlic, Grape, Tea, Berries,
Carrot,Spinach,Broccoli,
RedMeat,Kidney,Liver&LipoicAcid
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OxidativeStress: Undernormalconditionsbodymaintainsan
equilibriumbetweenitsownFRsand
Antioxidants.
Whenthisequilibriumbreaks,astatecalled
oxidationstressariseswithin,duetoFR
formationorAOsystem.

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Example:
AOStatusinNormalPregnancy: ThereisincreasedneedforAOsasthereisincreased
productionofFRsdueto
Pregnancy being a stressful condition.
Because of the rapidly growing foetus there is increased
cellular activity.

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 ThusAOactivityduringnormalpregnancy
progressivelyincreasesasdemonstratedby
Serum tocopherol.
Activity of GOP.
Serum ceruloplasmin & transferring level.

However,thereisnoevidencetosuggesttheneedfor
administrationofAOsduringnormalpregnancybut
drugsthatleadtoFRformationmustbeavoided.

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AOs&DiabeticEmbryopathy: Oxidativestresshasbeensuggestedtocontributeto
theincreasedriskoffoetalmalformationsinpoorly
controlleddiabetics.
Therearereportsof lipid peroxidation in cell
membranes indiabeticpregnancies
Periodsofmaternalhyperglycaemia&hypoglycemiamay
causemarkedchangesintheavailabilityofglucosetothe
foetus.
Alsoconc.oflipids,notablytheketonebodiesand
branchedchainaminoacidsinthematernalcirculation
contributetoalterednutritionfortheembryo

Leading to FR conc. & foetal malformation in

embryo.
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AOs&DiabeticEmbryopathy: Duringlaterpartofpregnancy
loadofglucoseinthemitochondriamayacceleratethe
flowofelectronsthroughrespiratorychainincluding
mitochondrialleakageoffreeradicals.
ThisleadstoproductionofFRinembryonictissuesto
causecongenitalmalformations.

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 Thusmaintenanceofnormalconcentration
ofmetabolitesofallnutrientclassmaybe
importantforpreventionofadversefoetal
outcome.
Butthequestioniswilldietary
supplementsaloneholdthekeytothe
futureindiabeticembryopathy?
However,maintenanceofbloodglucose
levelateuglycemiclevelisalwaysimportant
forpreventionofDiabeticembryopathy.
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AOs&DownSyndrome: Freeradicalsbeingthehallmarkofaging,are
greatlyincreasedwithmaternalage.
SoFRsplayaroleinpathogenesisofDowns
syndrome.
AdministrationofAOsmayhelpinpreventing
thisdisease.

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FRs&AOsinPIH: Increasedactivityoffreeradicalspromotematernal
uterinevascularmalformations.
FRs are promoters of maternal vasoconstriction.
O2 , H2 O2 & NO2 in combinations
InactivatetheNO(avasorelaxant)
CausesPGsynthataseactivity.
Produceperoxynitrate,apotentoxidant

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

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FRs&AOsinPretermDelivery: Subclinicalchorioamnionitisleadstoliberation
ofbacterialantitoxins(polysaccharidein
nature)andinflammatorycytokines.
Thesecausestimulationofinflammatorycells
andcellsofCX.whichinturnproduceNO2.
Evidence:ThereisN2Oinvaginalsamples
offemaleshavingpretermdelivery.
Soantioxidantsmayplayapartinthe
treatmentofpretermdelivery.

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FRs&AOsinNewBornBabies: Inrecentyearsexperimentalandclinicaldata
haveprovidedcompellingevidencesfor
involvementofoxygenderivedFRsin
disordersofprematurity.

Chroniclungdisease
Retinopathy
Intraventricularhaemorrhages.
NecrotisingEnterocolitis:-FRscause
microvascularinjuryandcellpermeabilityleading
tonecrotisingenterocolitis.

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FRs&AOsinNewBornBabies: PDA:-

HypoxanthineradicalinNBBleadstoformation
ofPGE2andcausesPDA.

Kwashiorkor: Studiessuggestthatkwashiorkorbabieshave
FRsintheirbody.
AOinKwashiorkorbabiesareduetodeficiency
ofVit.A,E,C,Inositol&Selinium.

IdeallythefuturetreatmentofNeonateswith
allthesediseaseshouldincludeAOtherapy.

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Carcinogenesis: Basicsofcancerformation: Anormalcellcanundergomalignant

transformationinpresenceofprocarcinogensand
carcinogens.
However,inearlystages,itcanrevertbackto
normalcellwhendetectedandcorrectedbyour
bodyimmunesystem.
Withoutimmunesystemdetectionanormalcellis
convertedtomalignantcellinstages
InitiationPromotionProgression

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FRsinCarcinogenesis: Freeradicalsareformedfromstimulantslike

Radiation
-Xenobiotics
-Inflammatorycells
-Respirationetc,
whichactoncellulartargetstocauseoxidant
DNAdamageinformofmutagenesis&
clostogenesis,

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-whichcause:-initiationofcarcinogenesisby Activationofprotooncogens.
Inactivationorlossoftumoursuppressergenes.
Normalcellbecomesinitiatedcell.

ProcarcinogensaremetabolicallyactivatedbyFRs,
whichcausepromotionandprogressionofthese
initiatedcellstocancer
Ultimatelycarcinogenesissetsin

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FRs&AOsinCarcinogenesis:FRsActivationof
procarcinogens&
Carcinogens
GeneticDamage

FRs

Normalcell
Initiation

RepairbyAOs

InitiatedCell
Promotion

RepairbyAOs

- Tumourpromoters
- Spontaneous

FRs
- Dysregulation
- GreaterCellautonomy
- ReducedGrowth
Factordependence

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PremalignantCell
RepairbyAOs

Progression

MalignantCell

CLINICAL CANCER
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AOsinCancerPrevention: DEFENCE:-enzymeswithantioxidant
propertycausefirstlineofdefenceby: Protectingthelipidsandenzymesagainstoxidation
Retardingthegenerationoffreeradicals
CreatingabalanceofAOsagainstFRsinthe
bodyPREVENTS:

Cellpathology.
Metabolicdisturbances.
Changesincellpermeability.
Eormationoftoxicproducts.

PreventionofinitiationofCarcinogenesis
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AOsinCancerPrevention: INTERVENTIONatpromotion&progression
stages:
Localdeactivationofgenotoxinsresponsiblefor
furthernuclearmutations
Inactivationoftumourpromoterseg.-activationof
granulocytes.
Simulateoxygen.
Maintenanceofproperfunctionofgapjunction
communication.
Maintenanceofphysicalstabilityofmembrane&
alsowithincells.
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AOsinCancerPrevention: Majorityofepidemiologicaldatasuggest
supplementationwithantioxidants
-Vit.-A,E,C;
-betacarotene&selenium,decreasesthe
incidenceofvariouscancers.

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AOsinCancerPrevention:RECOMMENDED FOR CARCINOMA PREVENTION

(Federation of Obstetric & Gynaecological Societies of India)
Antioxidant

RDA

Recommended Possible
Dose
ToxicityLevel

VitaminA

5000IU

12,500IU

Chronicintake Smoking
of125,000IU

VitaminE

10-20IU

200-800IU

>1,200IU

HighPUFAintake,
Smoking

VitaminC

60mg

1000mg

Negligible/12Gms

Stress,
OCP,Smoking

Selenium

None

50-200mg

>200mg

Aging,HighPUFA
intake,Smoking,
Heavymetals

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FeaturesCausing
Req.

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FRs&AOsinInfertility-Male
VariousstudieshavesuggestedConc.of
Malondi-Aldehyde(MDA)isinversely
proportionaltofertilityincaseofmales.
Inasthenospermicandoligoasthenospermic
malesthereisincreasedserumconcentration
ofMDA.
Eveninnormospermicmalesifthereis
concentrationofMDAthereisreducedfertility.
AdditionofvitaminEcausesdecreased
concentrationofMDAandimprovesfertility.
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FRs&AOsinInfertility-Female
FRscause ShortlutealphaseandLPDS.
InIVFarrestthecellgrowth(div)at2,4,8cell
stages
Hampertheregulationofcorpusluteum.

AdditionofAOsimprovestheresults.

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CONCLUSION
Oxidantgenerationisapartofhumanlife.
EveryO2atomwebreathin,isconvertedtowater
insideourbodybyadditionoffourelectrons
sequentially.
WhenfourelectronsareaddedthreeFRs-O2,
H2O2&OHareformedalongwithwater.
Sowherethereislifethereareoxidants.
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CONCLUSION
Butwhenproducedinexcess,theycancause
anydisease.
Soourconcernshouldbeto FRs in systemic
circulation.
However,carefuluseofAOsandnewerand
moreaccuratemethodstomeasureoxidant
generationinhumans,willgoalongwaytofind
outtheexactcontributionofoxidantsindisease
processesandtheroleofAOstopreventit.
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THANK YOU
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