ORIGINAL ARTICLE
The Influence of Calorie Restriction During Fasting
of The Month of Ramadhan on Free Radicals and Antioxidants
Expressed in The Form of Malondialdehyde and Glutation
in Healthy Young Males
Kriscahoyo D*, Makmun LH**, Setiati S**, Dharmeizar***, A.Jusman SW****
ABSTRACT
Aim: To observe the effect of calorie restriction on the
formation of lipid peroxide (rnalondialdelzyde/MDA)caused
by free radicals and antioxidant level (glutathioize/GSH)
among healthy young males dnring Ramadan fasting.
Subjects and Methods: Prospective ser-contl-olled study
was peiformed on 22 healthy men aged 20-25 years who
were presumed to be undergoing the calorie restriction.
Subjects were observedfour times: day 7priorto the Ranzadan
month, the 7'" and 21" days of Ramadan and day 7 after
Ramadan. All data are presented as mean averages (SEM);
all statistical comparisons were made using Anova Repeated
Measures and Pearson's bivariate correlation.
Results: Caloric intake decreased by 22 - 25% and
MDA decreased signiJicantly (p= 0.002), while GSH levels
increased signc3cantly (p= 0.047). There was no correla-
tion between calorie restriction and MDA levels (p= 0.27;
r= 0.2% but calorie restriction and GSH were correlated
(p= 0.049; r= -0.18).
Conclusion: This study showed that calorie restriction
during Ramadan fasting decreased MDA and raised plasma
GSH levels.
* Department of infernal Medicine, Faculfy of Medicine of ;Ire
Universiry of hdonesio,Dr. Cipto Mnngunkusumo National Central
General Hospital, Jakarta
*" Division of Geriatrics, Departnrnlmr of internal Medicine, Fnculq of
Medicine of the University of Ixdonesia, Dr. Cipro Mnngu,tkrcssmo
National Central General Hospital, Jakarta
*** Division of Renol-Kyperrerrsion,Depnrfmentof fnte,nol Medicine,
Faculty of Medicine of the University of lndortesio,Dr. Cipto
Mangunkusunm Natioxol Cenfrul General Hospital, Jokartu
**** Deparhnerrt of Bioclrenrirtq~Faculry of Mediciw of the Universio~
of Indonesia.
Volyme XXXV Number 4 October-December 2003
BACKGROUND
The free radical hypothesis plays a role in the
pathogenesis of man disease^,',^,^ such as heart
disea~e,l,~ diabetes
. ~ - ~ m e l l i t ~ s , ' atheros~lerosis,'~-'~
~~~~~'~
h y p e r t e n ~ i o n , ' ~rheumatoid
~'~ arthriti~,'.'~ respiratory
di~ease,'.~"'~ ~ a t h a r a c t , ' liver
~ ~ ~ fatty
~ ~ ' infiltra-
t i ~ n , ' infertility,34-37
~-~~ and early aging.'845
The presence of cell injury due to free radicals is
characterized by the formation of lipid p e r o ~ i d e . ~ "This ~~~'
process of fat degradation will produce malondialdehyde
(MDA). This substance could also be found in the blood,
so that it is often used as an indicator of the presence of
cellular and tissue damage due to free radi~aIs.4~
The body's mechanism to deal with such condition
is by producing GSH antioxidant^.'^"^ This substance is
considered as an impoaant antioxidant, since it is able to
detoxify lipid peroxide, water solubleperoxide,YO,, lipid
hydrogen peroxide within the cell membrane, various drug
metabolites, pollutants, heavy metals, and maintain
T lymphocyte function. Vitamin C and E activity also
depend on GSH.54
Calorie restriction studies on laboratory animals
demonstrated longer age span and a condition free from
disease. The study by Weindruch demonstrated a
reduction in free radical^.""^ Studies on hwnans are still
very limited, due to human's longer age span and
diiculty to isolate. The Ramadan fasting month is the
best way to use as a model of calorie re~triction.~~ Young
subjects are expected to be able to achieve a calorie
restriction similar to that among laboratory animals,
ranging around 30%.
 The Influence of Calorie Restriction During Fasting of The Month of Rarnadhan on Free Radicals and Antioxidants
Expressed i n The Form of Malondialdehyde and Glutation in Healthy Young Males
Figure 3. Mean Average Vitamin C, Vitamin E and Beta Carotene
Intakes During Fasting of The Month of Ramadhan
% OMDA (nmoUml)
5 week, while the BMI remains within normal limits. This
2=
5 reduction could be found in Figure 6.
Figure 4. Mean Average Calorie Restriction MDA Levels and GSH
During Fasting of The Month of Ramadhan
DISCUSSION
The calorie restriction achieved in this study was
25.23% on the third point of observation. This value is
greater than that compared by the studies by Setiati
(12.29%),44Deden (9.3%),55Julwan (12.15%),56and
Haricahyo (10.37%) who used elderly subject^.^' Such
results approximates the degree of calorie intake restric-
tion found among laboratoly animals, of around 30%. To
clarify this calorie restriction, we could see a degree of
reduction in bodyweight and body mass index (BMI).
There is a reduction of approximately 2 kg or 2.7% of
the original body weight, but the BMI was still within
normal limits of 20.36. This could be found in Figure 5.
These results are in accordance with the study by Seileh,
Mustafa, and A1-Habba1.58,60,6i
Figure 5. Mean Average Change in Bodyweight and BMi Durlng
Fasting of The Month of Rarnadhan
Volume XXXV . Number 4 October-December 2003
The level of MDA achieved during the Ramadan
fast is 107 SE 9.74, lower than that found by Setiati
(229.21 SE 24.25). The difference is in keeping with the
study by Nutall, where MDA levels are increased in the
elderly?0 The study limitation is the limited sample size
and the absence of a control group.
The Influence of Ramadan Fasting on Calorie
Restriction
Fasting during the month of Ramadan still ensures a
balanced diet, both in quality and quantity, and is instead
more directed towards a reduction in total quantum
energy or calorie.59In this study, calorie restriction is
22.13% during the first week and 25.23% in the last
could be found in Figure 3. The significant trace element
--iF
0Selenium
-"; 58$l.76 0.6f z i n c
27 36 28 +Cuprum
r 0
I n m IV
Okvation
Figure 6. Mean Average Selenium, Zinc and Cuprum Intake During
Fasting of The Month of Ramadan
The Influence of Calorie Restriction on MDA and GSH
Levels
Laboratory animal studies ascertain that calorie
intake restriction reduces the level of free radicals. What
is meant by calorie intake restriction in this case is
reduction of total calorie from carbohydrate, protein, and
lipid intake.
As explained by Weindruch, several mechanisms that
reduce the level of free radicals and increase
antioxidant levels are: reduced metabolite production rate,
increased glucose 6 phosphatase due to gluconeogen-
esis, and reduced xantin o x i d a ~ e . ~ ~ . ~ ~
In this study, MDA levels are significantly reduced
during fasting (p=0.002), and GSH levels are signiticantly
increased (p=0.047). The results can be found in Figure
4. These results are in accordance with studies on
hyperglycemic states, as conducted by Loven and other
researchers, where such condition significantly increases
MDA and reduces GSH, while SOD and catalase
demonstrate controversial results.12
Bivariate Analysis of Calorie Intake Restriction with
MDA and GSH Levels
During fasting, there was significant calorie intake
restriction (p=0.000), significant MDA reduction
 The Influence of Calorie Restriction During Fasting of The Month of Ramadhan on Free Radicals and Antioxidants
Expressed in The Form of Malondialdehyde and Glutation in Healthy Young Males

Figure 12. Correlation Between Beta Carotene and MDA Levels Figure 15. Correlation Between Beta Carotene and GSH Levels

CONCLUSION
Calorie intake restriction during the Ramadan
fasting reduces MDA levels and increases GSH levels.
There needs to be further studies using larger
samples and control groups.
There needs to be further studies examining
exogenous plasma antioxidant levels.
Calorie intake restriction could possibly be
considered as an alternative to increase endogenous
-
0Observed antioxidant levels amidst the controversial use of
'1 Linear supplementary antioxidants.
0 Calorie intake restriction is very much recommended
Figure 13. Correlation Between Vitamin E and GSH Levels
as a p x t of a healthy lifestyle.

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