Effect of Palm Oil and Nigella Sativa in Aorta and Coronary Arteries of Albino Rats

ZAFAR IQBAL, MOHAMMED AHMED

ABSTRACT
Background: Fat remains a hot topic because of concerns over associations beteen consumption of
fats an! the inci!ence of some chronic con!itions inc"u!in# coronar$ arter$ !isease, !iabetes,
cancer an! obesit$% Dietar$ pa"m oi" in ba"ance! !iets #enera""$ re!uce! b"oo! cho"estero", "o&
!ensit$ "ipoprotein 'LDL( cho"estero", an! tri#"$ceri!es hi"e raisin# the hi#h&!ensit$ "ipoprotein
'HDL( cho"estero"% )a"m oi" is en!oe! ith a #oo! mi*ture of natura" antio*i!ants an! to#ether
ith its ba"ance! composition of the !ifferent c"asses of fatt$ aci!s, ma+es it a safe, stab"e an!
versati"e e!ib"e oi" ith man$ positive hea"th an! nutritiona" attributes%
Obective: ,o stu!$ the effect of pa"m oi" an! -i#e""a sativa on the aorta an! coronar$ arteries of
a"bino rats%
!aterials and !et"ods. /i*t$ a"bino rats inc"u!in# e0ua" number of ma"es an! fema"es ere obtaine!
from )1/IR Laboratories, Lahore% After overni#ht fastin# of 23&24 hour a"" the anima"s ere ei#he!
an! 5ero an! 34 ee+s% ,he rats ere !ivi!e! into five #roups of te"ve anima"s each% Each #roup ha!
si* ma"es an! si* fema"es +ept in separate ca#es% At this sta#e the te"ve anima"s of #roup 16 ere
sacrifice! as the contro" #roup for a"" the other #roups to compare a#e chan#es% ,hen the e*perimenta"
!iet as starte! for the remainin# four #roups "abe"e! as 12, E2, 13 an! E3 base! on the !iets fe!. 12
'contro" #roup fe! on 78 pa"m +erne" oi"(, E2 'e*perimenta" #roup fe! on 78 pa"m +erne" oi" ith
-i#e""a sativa(, 13 'contro" #roup fe! on 368 pa"m +erne" oi" ith cho"ic aci! an! thiouraci" aci!( an!
E3 'e*perimenta" #roup fe! on 368 pa"m +erne" oi" ith cho"ic aci!, thiouraci" aci! an! -i#e""a sativa(%
First samp"e as co""ecte! as base"ine before startin# the e*perimenta" an! contro" !iets '6 ee+( an!
the secon! b"oo! samp"e as co""ecte! at the en! of the e*periment i%e% after 34 ee+s% ,o cc b"oo!
as !ran from each rat% At 34 ee+s, after co""ectin# b"oo! samp"es, the rats ere p"ace! in an ether
9ar ti"" !eath% ,he ab!omina" an! thoracic cavities ere opene! throu#h a me!ian incision% ,he hearts
an! aortae ere c"eare!% Biops$ specimens ere +ept in "abe"e! 9ars for fi*ation containin# 268
neutra" buffere! forma"in% ,hen the specimens ere sub9ecte! to #ross an! microscopic e*amination%
Results. :ei#ht #ain as statistica""$ si#nificant hen e*perimenta" #roup E2 as compare! ith
contro" #roup 12 an! hi#h"$ si#nificant hen e*perimenta" #roup E 3 as compare! ith contro"
#roup 13 % :hen the aortae ere opene! ith there ere neither fatt$ strea+s nor atheromatous
p"a0ues in the aortae of a"" e*perimenta" an! #roup anima"s% -arroin# of coronar$ arteries as not
observe! in an$ of the #roups% % )a"m oi" increase! the tota" cho"estero" an! "o !ensit$ "ipoprotein
cho"estero", !ecrease! the hi#h !ensit$ "ipoprotein cho"estero" "eve"s in a"" #roups at 34 ee+s as
compare! to 6 ee+ samp"es% -i#e""a sativa !ecrease! serum tota" cho"estero" an! "o&!ensit$
"ipoprotein cho"estero" an! increase! hi#h !ensit$ "ipoprotein cho"estero" "eve"s hich ere hi#h"$
si#nificant%
Conclusion. A"thou#h there as si#nificant ei#ht #ain an! e"evation of tota" cho"estero" an! "ipi!s,
these fai"e! to pro!uce atherosc"erotic "esions in the e*perimenta" rats on )a"m +erne" oi" in 34 ee+s
#ey $ords. Aorta, 1oronar$ arteries, A"bino rats, )a"m +erne" oi", -i#e""a sativa
%NTRO&'CT%ON
,he impact of an e*cess of fatt$ aci!s in the !iet on car!iovascu"ar !iseases has been stu!ie! an!
!iscusse! both in human an! anima" stu!ies% ;enera""$, e*cessive saturate! fats increase the ris+, hi"e
unsaturate! fats are consi!ere! "ess harmfu" % It is e"" estab"ishe! that p"ant pheno"ics e"icit various
bio"o#ica" activities, ith positive effects on hea"th% )a"m oi" pro!uction resu"ts in "ar#e vo"umes of
a0ueous b$&pro!ucts containin# pheno"ics% :ater&so"ub"e pheno"ics from the oi" pa"m possess
si#nificant bio"o#ica" properties% )a"m oi" is enriche! in vitamin E in the form of a"pha&, #amma&, an!
!e"ta&tocotrieno"s% Dietar$ tocotrieno" supp"ements have been shon to prevent atherosc"erosis
!eve"opment in patients an! prec"inica" anima" mo!e"s% ,he "in+ beteen !ietar$ fats an!
car!iovascu"ar !isease has create! a #roin# interest in !ietar$ re! pa"m oi" research% A"so, the "in+
beteen nutrition an! hea"th, o*i!ative stress an! the severit$ or pro#ression of !isease has stimu"ate!
further interest in the potentia" ro"e of re! pa"m oi" 'a natura" antio*i!ant pro!uct( to improve o*i!ative
status b$ re!ucin# o*i!ative stress in patients ith car!iovascu"ar !isease, cancer an! other chronic
!iseases% O*i!i5e! unsaturate! fatt$ aci!s ma$ contribute to the patho#enesis of atherosc"erosis% ,he
pa"m fruit 'E"aies #uineensis( $ie"!s pa"m oi", a pa"mitic&o"eic rich semi so"i! fat an! the fat&so"ub"e
minor components, vitamin E 'tocophero"s, tocotrieno"s(, carotenoi!s an! ph$tostero"s% /tu!ies have
foun! that -i#e""a sativa essentia" oi" is more effective in re!ucin# the e*tent of potassium bromate
in!uce! mu"tip"e or#an to*icit$ 'car!iac an! "iver en5$mes imba"ance( that i"" u"timate"$ he"pfu" in
re!ucin# the e*tent of m$ocar!ia" an! "iver necrosis% ,herefore, -/ is a potentia" protective natura"
a#ent a#ainst atherosc"erosis an! car!iovascu"ar comp"ications in these patients%
!ATER%A( AN& !ET)O&S
/i*t$ a"bino rats inc"u!in# e0ua" number of ma"es an! fema"es ere obtaine! from )1/IR
Laboratories, Lahore% At the start of the e*periment, the$ varie! from 23&24 ee+s of a#e an! of 2<6&
3<6 #rams ei#ht% ,he$ ere +ept in anima"=s house of )ost#ra!uate Me!ica" Institute, Lahore% ,he
atmospheric temperature as +ept at 34>31 on an avera#e ith foo! an! ater avai"ab"e a"" times%
anima"s ,he anima"s ere ei#he! at 5ero an! tent$ four ee+s% purifie! maintenance !iet% After
overni#ht fastin# of 23&24 hour a"" the anima"s ere ei#he! an! 5ero ,he rats ere !ivi!e! into five
#roups of te"ve anima"s each% Each #roup ha! si* ma"es an! si* fema"es +ept in separate ca#es% For
the i!entification of the in!ivi!ua", the ear ho"e metho! of mar+in# as a!opte! 'Arrin#ton 2?@A(% A
rat ear punch as use! for this purpose% Before the start of e*perimenta" !iets, a four ee+s
acc"imati5ation perio! as #iven !urin# hich the$ fe! an! b"oo! samp"es ere ta+en% At this sta#e
the te"ve anima"s of #roup 16 ere sacrifice! as the contro" #roup for a"" the other #roups to compare
a#e chan#es% ,hen the e*perimenta" !iet as starte! for the remainin# four #roups "abe"e! as 12, E2, 13
an! E3 base! on the !iets fe!. 12 'contro" #roup fe! on 78 pa"m +erne" oi"(, E2 'e*perimenta" #roup fe!
on 78 pa"m +erne" oi" ith -i#e""a sativa(, 13 'contro" #roup fe! on 368 pa"m +erne" oi" ith cho"ic
aci! an! thiouraci" aci!( an! E3 'e*perimenta" #roup fe! on 368 pa"m +erne" oi" ith cho"ic aci!,
thiouraci" aci! an! -i#e""a sativa(% First samp"e as co""ecte! as base"ine before startin# the
e*perimenta" an! contro" !iets '6 ee+( an! the secon! b"oo! samp"e as co""ecte! at the en! of the
e*periment i%e% after 34 ee+s% ,o cc b"oo! as !ran from each rat% 1"ean an! !r$ test tubes ere
"abe"e! for each samp"e an! stan!ar! 6%6< m" '<6 B"( of each samp"e an! stan!ar! ere ta+en an! put
into tubes "abe"e! CtestD an! Cstan!ar!D% 3%6 m" '3666 B"( of su"phuric aci! as a!!e! to the tubes% ,he
tubes ere mi*e! e"", p"u##e! ith cotton oo" an! a""oe! to stan! in boi"in# ater for 26 minutes
then coo"e! in aci! ater for < minutes 6%2 '266 B"( of so"ution as ta+en from CtestD an! Cstan!ar!D
"abe"e! tubes% 6%2 m" '266 B"( of su"phuric aci! as ta+en into a tube "abe"e! as b"an+% 6%3 m" '366 B"(
of co"or rea#ent as a!!e! into a"" tubes% A"" tubes ere a""oe! to stan! for 76 minutes at 7@1 an!
tests ere measure! a#ainst rea#ent b"an+ at <66 nm usin# Lab s$stem 1hemistr$ Ana"$5er F)&?62%
Mean va"ues of "ipi! profi"es in a #roup at 6 an! 34 ee+s an! beteen !ifferent #roups at 34 ee+s
ere compare! ith the he"p of stu!ent=s Et= test% At 34 ee+s, after co""ectin# b"oo! samp"es, the rats
ere p"ace! in an ether 9ar ti"" !eath% ,he ab!omina" an! thoracic cavities ere opene! throu#h a
me!ian incision% ,he hearts an! aortae ere c"eare!% Biops$ specimens ere +ept in "abe"e! 9ars for
fi*ation containin# 268 neutra" buffere! forma"in% ,hen the specimens ere sub9ecte! to #ross an!
microscopic e*amination% Each aorta as ro""e! on itse"f ith the intima" surface our, startin# ith the
thoracic en! hi"e the hearts ere opene! transverse"$, #ross"$ e*amine! an! 3 mm thic+ s"ices ere
cut for processin#% ,he processin# of tissues as !one in an automatic processor% ,he paraffin b"oc+s
ere ma!e in L&shape! mou"!s% /ections of 7&4um thic+ness ere cut b$ rotar$ microtome% ,he
sections ere ta+en on a"bumeni5e! s"i!es% ,he sections ere staine! ith hemato*$"in an! eosin, van
;ieson, Ferhoeff=s e"astic tissue, oi" re! o amon# other stains%
Results
Difference of ei#ht #ain at 34 ee+s as compare! ith contro" an! re"evant e*perimenta" #roups%
,his as statistica""$ si#nificant hen e*perimenta" #roup E2 as compare! ith contro" #roup 12
an! hi#h"$ si#nificant hen e*perimenta" #roup E 3 as compare! ith contro" #roup 13 'tab"e 2(%
:hen the aortae ere opene! ith there ere neither fatt$ strea+s nor atheromatous p"a0ues in the
aortae of a"" e*perimenta" an! #roup anima"s% -arroin# of coronar$ arteries as not observe! in an$
of the #roups% Microscopic e*amination of sections from aorta an! coronar$ arteries of !ifferent #roups
at 34 ee+s as !one an! these revea"e! norma" histo"o#$ both in the contro" #roup an! e*perimenta"
#roup% /pecifica""$ in a"" #roups no /tar$=s "esion as seen, van ;ieson stain revea"e! norma" pattern
of co""a#en% Ferhoeff=s stain revea"e! norma" pattern an! !istribution of e"astic fibers% -one of the
aortae an! coronar$ arteries in an$ #roup shoe! intrace""u"ar or e*trace""u"ar subinitma" "ipi! on oi"
re! O stainin#% ,he mean serum tota" cho"estero", serum tri#"$ceri!es, tota" serum "ipi!s amon# the
contro" an! e*perimenta" #roups are shon in tab"e 3
&iscussion
Atherosc"erotic !isease 'AD( is the "ea!in# cause of !eath or"!i!e% /outh&Asians '/As( sho an
increase! ris+ of atherosc"erosis an! have the hi#hest morta"it$ rates from 1AD than an$ other ethnic
#roup% ,he "ea!in# in!epen!ent pre!ictors of coronar$ events an! !eath Gs$sto"ic b"oo! pressure,
tota"HHDL&cho"estero" ratio, fo""oe! b$ !iabetes an! 'centra"( obesit$I are re"ate! to the metabo"ic
s$n!rome, estimate! to prevai" in 7&48 of a!u"ts a#e! 76 or over, an! to un!er"ie one&ei#hth of cases
of coronar$ !isease%
,he increase in ei#ht that occurre! in rats fe! hi#h fat !iet has been foun! in other stu!ies as e""% -o
aortic p"a0ues ere foun! in the stu!ie! rats, this as true across a"" the #roups& contro""e! or
uncontro""e!% ,his fin!in# has a"so been substantiate! b$ other stu!ies, hich !i! not fin! the
occurrence of these p"a0ues after J&23 ee+s of pa"m oi" % ,he beneficia" effect of pa"m oi" on coronar$
arteries is e"" +non%
It is true that hi#her "eve"s of ,1 an! LDL&c ere obtaine! hich p"a$ a positive ro"e in athero#enesis
but at the same time, HDL&c "eve" as a"so raise! hich has a protective ro"e in the #enesis of
atherosc"erosis% ,his has been foun! in the or+ !one b$ other co""ea#ues% Other or+ers have shon
that !ifferent !ietar$ components can beneficia""$ mo!u"ate free ra!ica" me!iate! o*i!ative stress
in!uce! b$ "ipi! pero*i!ation% ,he$ a"so !emonstrate! that pa"m oi" consumption re!uces b"oo!
cho"estero" in comparison ith the tra!itiona" sources of saturate! fats such as coconut oi", !air$ an!
anima" fats% In a!!ition, pa"m oi" consumption ma$ raise HDL "eve"s an! re!uce p"ate"et a##re#abi"it$%
Ramachan!ran an! other or+ers foun! that serum "ipi! ana"$sis shoe! si#nificant increase in
cho"estero", LDL&c K FLDL&c an! !ecrease in HDL&c "eve"s in a"" the treate! #roup of anima"s%
,he a""e#ation that pa"m oi" consumption "ea!s to raise! b"oo! cho"estero" "eve"s an! is therefore
athero#enic is ithout scientific foun!ation% E*amination of the chemica" an! fatt$ aci! composition of
pa"m oi" or its "i0ui! fraction shou"! convince most nutritionists that the oi" has "itt"e cho"estero"&
raisin# potentia"% E"son an! other or+ers have foun! that a #roup of antio*i!ants, the tocotrieno"s, are
present in both pa"m o"ein an! re! pa"m oi"% ,hese vitamin E&active constituents are potent suppressors
of cho"estero" bios$nthesisL emer#in# !ata point to their anticarcino#enic an! antithrombotic activities%
It has been shon that pa"m oi", obtaine! from a tropica" p"ant, E"aeis #uineensis contains <68
saturate! fatt$ aci!s, $et it !oes not promote atherosc"erosis an! arteria" thrombosis% ,he saturate! fatt$
aci! to unsaturate! fatt$ aci! ratio of pa"m oi" is c"ose to unit$ an! it contains a hi#h amount of the
antio*i!ants, beta&carotene, an! vitamin E%
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Table *:Com+arision of mean,- S& body .eig"t /grams0 in different grou+s at 1 and 23 .eeks
4rou+ !ale Rats
/1 .eek0
!ale Rats
/25 .eeks0
6emale
Rats /1
.eeks0
6emale
Rats /23
.eeks0
Total /1
.eeks0
Total /23
.eeks0
C* 21178,-*878 98*75,-**79 *:372,-*273 288,-*579 *8:7;,-2175 99;71,-;17;
E* 2127;,-*;73 92979,-2375 *5;71:7* 2:575,-57* *8978,-2275 91171,-2<7:
C2 21;78,-*:73 91;78,-875 *51,-*17; 2:979,-879 *827<,-2:75 28<7;,-*878
E2 21575,-*875 2<;71,-*272 *5879,-*179 251,-:73 *8:7;,-2375 2::,-2179
P=171*/"ig"ly significant0
Table 2: (i+id +rofile in different grou+s at 23 .eeks in mg>dl/!ean,-S&0
4rou+s Total
C"olesterol
)&( (&( Total
glycerides
Total li+ids
C* <37<,-379 2178,-57; ;179,-575 **87:,-579 33:72,-87:
E* <;75,-97; 2573,-379 3;7<,-37* *2*75,-37* 33:7*,-87<
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