The efect of cafeine on lipid peroxidation in the liver and kidney of

Wister Albino rats

Onwuka F
1

Erhabor O
2
epart!ent of "ioche!istry# $olle%e of "asic &edical 'ciences#
(niversity of )ort *arcourt
1
# epart!ent of *e!atolo%y $olle%e of
*ealth 'ciences (niversity of )ort *arcourt
2
+
All correspondence to
r Erhabor Osaro
1 "ra,ley Avenue "olton (-#
"./ 2.+
E0!ail1n2osaro3yahoo+co!
Abstract
1
Cahene (1, 3, 7-trmethy xanthne), a whte crystane xanthne akaod, s
a psychoactve stmuant drug. It s a common content of severa beverages
and has been documented to have many ehects on severa body organs and
systems. One of the mportant consequences of the consumpton of cahene
s ts ehect on pd peroxdaton, a cruca step n the pathogeness of severa
dsease states n humans. The present paper reports the resuts of an
nvestgaton of the ehects of cahene, gven as cohee or tea, on the
generaton of maondadehyde (MDA), a measure of pd peroxdaton, n the
ver and kdneys of wstar abno rats. The rats were dvded nto three
groups - a contro group of 9 rats whch receved norma feed and pan
water, and two other groups of 9 rats each whch receved 1 gm of ether
cohee or tea dssoved n 2 m of water n addton to ther norma feeds.
Treatment asted for 14 days after whch the anmas were sacrced and the
ver and kdneys harvested. The degree of pd peroxdaton, expressed by
the concentratons of MDA n extracts of the ver and kdneys, was assessed
n a rats. It was found that both cohee and tea ncreased the mean
concentratons of MDA n ver and kdneys of the rats. However, whe the
mean MDA concentraton n the ver was sgncanty hgher (p0.05) ony n
the rats that receved tea when compared wth the contro rats, t was
sgncanty ncreased (p0.05) n the kdneys of ony those rats that
receved cohee when compared wth the contro rats. It s concuded that
cahene, gven as ether tea or cohee, has a pro-oxdant ehect on the rat
ver and kdneys.
2
4ntroduction
The roe of cahene n pd peroxdaton s vared and possby depends on
the dose of cahene ngested and the anma speces studed. Cahene has
been recognzed as an emcent boogca ant-oxdant. However, severa
studes suggest that ths property s observed when cahene s present n
mmoar quanttes. Cahene was an ehectve nhbtor of pd peroxdaton,
at mmoar concentratons, n rat ver mcrosomes 1. Aso, Katyar et a
2
found that extracts of green tea had ant-carcnogenc and ant-nammatory
ehects n a mouse mode, propertes that correated drecty wth the
antoxdant propertes of the extracts of the tea. Grucker-Mamczar et a
3
aso reported that cahene, admnstered to rats n drnkng water at a
dosage of 3.1 mg/L emcenty nhbted uorde-nduced peroxdaton of pds
n hepatocytes. On the other hand, Danzan et a
4
(1991) found that cahene
s capabe of nducng oxdatve damage by fosterng pd peroxdaton n rat
ver. Azam and coeagues
5
studed the prooxdant actvty of cahene,
theobromne and xanthne by measurng the abty of each compound to
oxdatvey degrade DNA n the presence of copper ons. They found that the
three agents ed to ncreased oxdatve DNA breakage by hydroxy radcas
suggestng that cahene and ts metaboc products have prooxdant
propertes. Smary, Gcn
6
, workng wth an n vtro system found that
cahene promoted the peroxdaton of noec acd emusons by 32.5, 48.9,
3
and 54.3%, respectvey, at 15, 30 and 45 mcrog/mL concentratons n rat
vers.
Addtonay, t appears that, under certan stuatons, cahene shows no
postve or negatve ehect on pd peroxdaton. For nstance, Ocna and co-
workers
7
evauated the ehect of cahene on exercse-nduced oxdatve
stress n twenty mae human sub|ects n a doube bnd manner durng
maxma cycng exercse. They measured pasma vtamns A, E, and C as
markers of non-enzymatc antoxdant status, as we as maonadehyde
(MDA) as an ndex of pd peroxdaton and found that suppements of
cahene gven one hour before the exercse at a dose of 5mg/kg of body
weght, whe enhancng the ergosprometrc response to cycng, had no
ehect on pd peroxdaton or on the changes n the concentratons of the
antoxdant vtamns A, E and C, when compared to pacebo.
Thus, t appears that the observed ehect of cahene on peroxdaton s
varabe and probaby depends on the dose of the agent gven, the speces
studed, and the possbe presence of other agents ke avonods or
poyphenos that may be n the tea or cohee.
Ths study was therefore desgned to evauate the ehects of cahene, gven
as tea or cohee, on the process of pd peroxdaton n the rat ver and
kdney.
4
5
&aterials and !ethods
Cohee and Tea sod as Cassca Nescafe and Tope Tea were purchased from
Choba Market, Port Harcourt, Rvers State, Ngera. Twenty-seven adut Wster
Abno rats weghng between 175g and 325g were used for the experment.
The anmas were obtaned from the anma house of the Department of
Bochemstry of the Unversty of Port Harcourt. They were housed and kept
under aboratory condtons wth free access to a standard det and water.
The 27 rats were dvded nto three groups of nne rats each as foows; group
1 receved norma feed and pan water; group 2 rats were fed norma feed,
water and 1 gm of cohee dssoved n 2 m of water whe group 3 rats were
fed wth norma feed, water and 1 gm of tea dssoved n 2 m of water.
'a!ple collection and preparation: A anmas were sacrced after 14
days of treatment. They were anaesthetzed n a choroform saturated
chamber, one at a tme, n desccators; they were then sacrced whe st
under anesthesa. Each rat was dssected and the ver and kdneys of each
anma were excsed, separatey homogenzed and appropratey prepared for
the determnaton of tssue concentraton of maondadehyde (MDA) for the
assessment of the degree of pd peroxdaton. Tssue maondadehyde
concentratons were determned by the method of Hunter et al
8
as moded
by Gutterdge and Wkns
9
. A 1g sampe of each ver or kdney was
homogenzed n 9m sane souton. 1.2m of the sampe homogenate was
added to 6m of gaca acetc acd and 6m of 1% thobarbturc acd
6
dssoved n 0.2% NaOH souton n a test tube. The mxture was paced n a
water bath and aowed to bo for 15 mnutes, aowed to coo and was then
centrfuged at 2,500D for 10 mnutes. An aquot of the supernatant was
used to evauate the concentraton of MDA as an ndex of pd peroxdaton.
'tatistical Analysis
Data was anayzed usng a statstca package for soca scences SPSS
verson 10 (verson 10; SPSS Inc., Chcago, IL). Data were expressed as the
mean vaue standard devaton (SD) for each group. The resuts of the
contro group and each treatment group were compared usng the pared
Student t-test. P vaues of 0.05 were consdered to be statstcay
sgncant for a statstca comparson.
5esults
The resuts are shown n Tabes 1.1 to 1.4. Tabe 1.1 compares the mean
concentraton of MDA n the vers of the three groups of anmas. It shows
that the mean concentraton of maondadehyde n the ver of the rats n
Group 2 (treated wth Nescafe) and the mean concentraton of MDA n the
ver of the rats n Group 3 (treated wth Top Tea) were hghy sgncanty
greater than the concentraton of MDA n the Contro group. Tabe 1.2
compares the mean concentraton of MDA n the kdneys of the three groups
of anmas. The resuts show that the mean concentraton of MDA n the
kdneys of the test anmas treated wth Nescafe (Group 2) and the mean
concentraton of MDA n the kdneys of the anmas treated wth Top Tea was
sgncanty hgher than the mean concentraton of MDA n the kdneys of
the Contro anmas. In Tabe 1.3, the ehects of Nescafe on the concentraton
of MDA n the kdney and the ver were compared. The observed dherence
n the mean concentraton of MDA n the vers and the kdneys of the
anmas treated wth Nescafe was not statstcay sgncant. Smary, the
7
dherence n the mean concentraton of MDA n the vers and the kdneys of
the anmas treated wth Top Tea was not statstcay sgncant (Tabe 1.4).
Table 1+11 &ean &A levels in the .iver of experi!ental ani!als
GROUPS N MDA (unt/g tssue
10
-5
)
p-vaue
Group 1
(Contro)
9 2.50 0.24
Group 2
(Nescafe)
9 3.19 0.17 < 0.01
(1 & 2)
Group 3
(Top Tea)
9 3.35 0.19 < 0.01
(1& 3)
Table 1+21 &ean level of &A in the -idney of experi!ental ani!als
Groups N MDA (unt/g tssue
10
-5
)
p-vaue
Group 1
(Contro)
9 2.50 0.24
Group 2
(Nescafe)
9 3.01 0.20 < 0.01
(1 & 2)
Group 3
(Top Tea)
9 3.16 0.38 < 0.01
(1 &3)
Table 1+/1 $o!parison of the efects of $ofee on the &A levels in
the liver and kidney of experi!ental ani!als+
Group Organ n MDA
(unt/g tssue 10
-5
)
p vaue
(Treated vs. Contro)
Group 2
(Nescafe)
Lver 9 3.19 0.17
Group 2
(Nescafe)
Kdney 9 3.01 0.20 > 0.1
8
Table 1+61 $o!parison of the efect of treat!ent with tea on the
concentration of &A in the liver and kidney of experi!ental
ani!als+
Group Organ n MDA
(unt/g tssue 10
-5
)
p vaue
Group 3
(Top Tea)
Lver 9 3.35 0.19
Group 3
(Top Tea)
Kdney 9 3.16 0.38 > 0.2
iscussion
The resuts of the present study show that, the ngeston of cahene by rats
n the form of cohee or tea, gven at a dose of 1 g of cohee or tea n 2 m of
water, promoted pd peroxdaton as ndcated by an ncrease n the
formaton of maondadehyde (MDA) n the ver and kdneys of the rats more
than pacebo. Danzan et a
4
have aso reported ncreased peroxdaton n
the vers of cahene-treated rats. These resuts suggest that n ths
expermenta mode and desgn, cahene, provded as cohee or tea promoted
pd peroxdaton n the rat ver and kdneys snce MDA s an end product of
the process.
Athough t has been suggested that tea eaves tend to contan more cahene
than cohee beans t s we-known that a typca servng (.e. a cup) of tea
contans much ess cahene than a cup of cohee probaby because tea s
usuay brewed weaker for consumpton. As a resut most pubshed studes
suggest that cohee generay contans more cahene than tea
10-13
. It s we-
known that the amount of cahene n cohee or tea depends on a number of
9
factors, ncudng the varety of cohee bean or tea eaf, where the pant s
grown, partce sze used, and the method and ength of brewng or steepng.
Aso wth tea, studes show that eaf ocaton on the tea pant, ahects content
of cahene n that eaf. A of these factors mght account for the wde
varabty of cahene n tea and cohee reported n the terature
10, 12, 13
. In a
survey of cahene and theobromne eves n Ngeran beverages, Eteng et a
14
found that the cahene content of Nescafe cohee was 93.66 8.91 mg/g
whe t was 37.22 5.34 mg/g for Lpton tea. Unfortunatey they dd not
study Top Tea. Despte these reported dherences n cahene content of
cohee and tea, t s of nterest that both agents produced smar degrees of
pro-oxdaton n the present study. Unfortunatey, the cahene content of the
Nescafe cohee and Top Tea that were used were not drecty measured to
assure equvaent content n the 1 gm of each substance used.
It s aso possbe that the pro-oxdant ehect observed coud be a resut of the
depeton or antagonsm of naturay occurrng antoxdants ke superoxde
dsmutase, cataase, or vtamns A and E among others, by the contents of
the agents gven n the tea or cohee, thereby permttng peroxdaton to
proceed wth reduced antagonsm. In a prevous study n rats, Danzan et a
4
found ncreased pd peroxdaton n the ver of cahene-treated anmas and
suggested that a decrease of the concentraton of vtamn E, whch was aso
observed n the treated anmas, mght contrbute to the pd peroxdaton.
Unfortunatey, we dd not assess the content of any of the known enzymatc
or non-enzymatc antoxdants n our rats. Prevous reports have suggested
10
that cohee exerts a suppressve ehect on hypergycema by mprovng nsun
senstvty, party due to reducng nammatory cytokne expresson and
mprovng fatty ver
15, 16
.
Hepfu modcatons that shoud be consdered n future studes of ths type
shoud ncude; an estmaton of the eves of antoxdants n the serum and
tssues of both contro and test anmas, an assessment of the concentraton
of cahene n the cohee and tea fed to the anmas, evauaton of the cohee
or tea for other chemcas that may have prooxdant propertes and
ncreasng the number of anmas n each group mght aso be of beneca.
We observed that there was no statstcay sgncant dherence n the ehect
of Nescafe cohee on pd peroxdaton as measured by the generaton of
MDA n ether the ver or the kdneys of the rats? Smary, the ehect of Top
Tea on the generaton of MDA n the vers and kdneys of the rats were
comparabe. These suggest that the prooxdant ehect of cahene n the oray
admnstered cohee or tea ahected these organs smary.
5eferences
1. Devasagayam TP, Kamat |P, Mohan H, Kesavan PC. Cahene as an
antoxdant: nhbton of pd peroxdaton nduced by reactve oxygen
speces. Bochm Bophys Acta. 1996; 13:63-70.
2. Katyar SK, Emets CA. Green Tea and Skn. Arch Derm. 2000; 136: 989-
994.
3. Grucka-Mamczar E, Zae|ska-Foka |, Chubek D, Kasperczyk S,
Blaszczyk U, Kasperczyk A, Swtochowska E, Brkner E. The nuence
11
of sodum uorde and cahene on the actvty of antoxdatve enzymes
and the concentraton of maondadehyde n rat ver. Fuorde. 2000;
42:102-109.
4. Danzan M, Muzo G, Bocca M, Canuto R. (1991). Lpd peroxdaton n
fatty ver nduced by cahene n rats. Int | Tssue React. 1991; 13: 79-
85.
5. Azam S, Had N, Khan NU, Had SM. Antoxdant and prooxdant
propertes of cahene, theobromne and xanthne. Med Sc Mont. 2003;
9:325-330.
6. Gcn I. In vtro prooxdant ehect of cahene. | Enzyme Inhb Med
Chem. 2008; 23: 149-152.
7. Ocna G|, Muoz D, Imn RM, Cabaero |, Maynar |I, Crdova A, Maynar
M. Ehect of cahene on oxdatve stress durng maxmum ncrementa
exercse. | Sports Sc Med. 2006; 5: 621-628.
8. Hunter FE, Gebck |M, Hohsten PE, Wensten |, Scott A. Sweng and
yss of rats ver mtochondra nduced by ferrous ons. |. Bo. Chem.
1963; 238: 828-835.
9. Gutterdge |MC, Wkns C. Copper dependent hydroxy radca damage
to ascorbc acd formaton of thobarbturc acd reactve products. FEBS
Lett. 1982; 137: 327-340.
10. Kapan E, Homes |H, Sapeka N. Cahene content of tea and
cohee. S Afr Med |. 1974; 8:510-511.
11. Bunker ML, McWams M. Cahene content of common
beverages. | Am Det Assoc. 1979; 74:28-32.
12. Stavrc B, Kassen R, Watknson B, Karpnsk K, Stapey R, Fred P.
Varabty n cahene consumpton from cohee and tea: possbe
sgncance for epdemoogca studes. Food Chem Toxco. 1988; 26:
111-118.
13. Pena A, Lno C, Svera MI. Survey of cahene eves n reta
beverages n Portuga. Food Addt Contam. 2005; 22:91-96.
12
14. Eteng MU, Eyong EU, Eka OU, Umoh IB, Ebong PE, Ettarh RR.
Cahene and theobromne eves n seected Ngeran beverages. Pant
Foods Hum Nutr. 1999; 54: 337-344.
15. Yamauch R, Kobayash M, Matsuda Y, O|ka M, Shgeoka S,
Yamamoto Y, Tou Y, Inoue T, Katagr T, Mura A, Horo F. Cohee and
cahene ameorate hypergycema, fatty ver, and nammatory
adpocytokne expresson n spontaneousy dabetc KK-Ay mce. J Agric
Food Chem. 2010; 12; 58(9):5597-60.
16. van Dam RM. Cohee consumpton and the decreased rsk of
dabetes metus type 2. !ed "i#d$chr %e&ee$'d. 2006; 150():1821-
1825.
13