Babandi et al.

, (2017) BJMLS, 2(1):

OXIDATIVE STRESS INDICES AND CALCIUM LEVEL AMONG HYPERTENSIVE

PATIENTS IN KANO-NIGERIA
1*
Babandi, A.,2Muhammed, I.Y. 1Murtala, Y.,1Ibrahim, A.A.,1Madugu, A. U.,
1
Babagana, K.,3Jobbi, Y.,1Mashi, J.A., 1Yakasai, H.M., 1Shehu, D. and 1Abubakar, S.M.
1
Department of Biochemistry, Bayero University, PMB 3011, Kano-Nigeria
2
Department of Chemical Pathology, Bayero University and AKTH, PMB 3011, Kano-
Nigeria
3
Department of Hematology, Aminu Kano Teaching Hospital, PMB 3452, Kano-Nigeria
*Correspondence:ababandi.bch@buk.edu.ng /+2348086303170
ABSTRACT
Background: Hypertension is a multi-factorial disorder in which various physiological
mechanisms participate to elevate Blood Pressure (BP). When imbalance between the
production and elimination of reactive oxygen species (ROS) occurs, leading to
accumulation of free radicals in cells, oxidative stress may result. It has been implicated in
the pathogenesis and complications of hypertension and other diseases. It promotes
atherosclerosis through various mechanisms.The interaction between the ROS and
endothelium-derived nitric oxide (NO) thereby inhibiting vasodilation and hence
hypertension, is an important mechanism.This preliminary study was aimed at evaluating
the oxidative stress indices and serum calcium among hypertensive patients attending
Aminu Kano Teaching Hospital, Northwest Nigeria.
Methods:The levels of serum malondialdeyde (MDA) as biomarker of oxidative stress,
antioxidant vitamins and serum calcium were evaluated using standard methods in
hypertensive as well as normotensive volunteers attending Aminu Kano Teaching
Hospital. A total of four hundred (400) volunteers (200normotensives and 200
hypertensive patients) were enrolled for the study.
Results: Results indicated a significant decrease (p<0.05) in serum antioxidant
vitamins(C and E) and calcium level in hypertensive patients compared to normotensives,
while significant increase (p<0.05) inserum MDA level was observed in hypertensive
subjects compared to the normotensive volunteers.
Conclusion: This study suggests the increase of oxidative stress marker (MDA) in
hypertensive patients with concomitant decreaseof the antioxidant vitamins and calcium
leveland thus, disturbs oxidant-prooxidant equilibrium and hence may contribute to
complications of hypertension.
Key words:Hypertension, Oxidative stress,Antioxidant vitamins, Calcium, Kano

INTRODUCTION visual problems are observed. So, the

Hypertension is one of the major public assessment on the risk factors which
health problems and among the leading contribute more to development of blood
causes of death globally. Itdoubles the risk pressure, myocardial infarction, left
of cardiovascular diseases including ventricular hypertrophy, congestive heart
coronary artery disease, congestive heart failure, aneurysm, stroke, as well as chronic
failure, ischemic and hemorrhagic stroke, kidney disease (hypertensive nephropathy)
renal failure and peripheral arterial disease and hypertensive retinopathy(Kaori et al.,
(Earl et al., 2015).It is a ‘silent killer’ as it is 2006; Manciaet al., 2007), at an early stage
asymptomatic until its effects like stroke, to control them will prevent the health
myocardial infarction, renal dysfunction or hazards of hypertension (Kaori et al., 2006).
Citation: 1*Babandi, A.,2Muhammed, I.Y. 1Murtala, Y.,1Ibrahim, A.A.,1Madugu, A. U., 1Babagana, K.,3Jobbi,
Y.,1Mashi, J.A., 1Yakasai, H.M., 1Shehu, D. and 1Abubakar, S.M A(2017). Oxidative Stress Indices And Calcium
Level Among Hypertensive Patients In Kano-Nigeria BJMLS. 2(1):
 Oxidative Stress Indices And Calcium
The complications of hypertension are
related to either sustained elevations of
blood pressure, with consequent changes in
the vasculature and heart, or to the
accompanying atherosclerosis that is
accelerated by long-standing hypertension
(Sabriet al., 2004).Oxidative stress is well
known to be involved in the pathogenesis of
lifestyle-related diseases, including
atherosclerosis, hypertension, diabetes
mellitus, ischemic diseases, and
malignancies. It is known to be harmful due
to oxygen free radicals attack of biological
molecules such as lipids, proteins, and DNA.
However, oxidative stress also has a useful
role in physiologic adaptation and in the
regulation of intracellular signal
transduction. Therefore, it may be
considered as “a state where oxidative forces
exceed the antioxidant systems due to loss of
the steady balance between them (Lushchak,
2014).
Similarly, disturbed calcium metabolism
may play a role in the patho-physiology of
hypertension. Ionized calcium (Ca) acts as
an intracellular second messenger in
excitation-contraction coupling in vascular
smooth muscle (VSM) cells. The free
intracellular calcium concentration
determines the tension in VSM cells, thus
contribute to peripheral vascular resistance
(PVR) which found in hypertension
(Sudhakaret al., 2002).
To the best of the authors’ knowledge,
astudy to evaluateoxidative stress,
antioxidant vitamins and calcium level
among hypertensive patients in this part of
the country has not been conducted. Thus, a
preliminary study will provide insight and
assist in the prediction of the outcome of
these diseases. This preliminary study was
aimed at evaluating the oxidative stress
marker, antioxidant vitamins and serum
calcium among hypertensive patients
attending Aminu Kano Teaching Hospital,
Northwest Nigeria.
MATERIALS AND METHODS
Study Site
Kano state is the most populous commercial
state in the northwestern Nigeria with a
population of 9,383,68 according to 2006
National Census. However, currently there is
believed that Kano state has a population of
about 14,000,000 people (Encyclopeadia
Britannica, 2017).
Kano State lies between latitude 13°N in the
North and 11°N in the South and longitude
8°W in the West and 100E in the East (Kano
Metro,2016). The participants were recruited
from Aminu Kano Teaching Hospital
(AKTH).
This facility is the most patronized Tertiary
Health Care hospital in Kano state,
northwestern Nigeria. Itreceives patients
from all over the northwestern region of
Nigeria.
Sample Size Determination
Fisher’s formula, (1998) was used to
determine the sample size.
n= Z2 P (1-P)/d2
Where; n=the desired sample size
z=95% confidence interval or 1.96
d=degree of precision usually set at 0.05
P= 0.5%
The prevalence of 50% was used
Experimental Design
A total of 400 volunteersrecruited from
Aminu Kano Teaching Hospital (AKTH)
participated in this research. The
volunteerswere grouped into two groups;
group I consist of 200hypertensive patients
and group II consist of 200 normotensive
volunteers.
Patients with systolic blood pressure above
140 mmHg or diastolic blood pressure above
90 mmHg, or the ones receiving
antihypertensive medications, non smokers
and of both sexes were considered as
hypertensive patients, and the remaining
constituted the control group. Smokers and
those refused to give their consent were
excluded from this research
The ethical approval to carry out this study
was obtained from the ethical committee of
Aminu Kano Teaching Hospital. The
provision of the Helsinki declaration was
respected at every step of the study.
 Babandi et al., (2017) BJMLS, 2(1):

Blood sample collection and preparation
After obtaining a written consent, 5ml blood
was withdrawn aseptically from the anti
cubital veins from each patient and control,
the samples were centrifuged at 3000rpm for
10minutes followed byserum collection for
further analysis.
METHODS
Serum calcium was determined by
enzymatic colorimetric method of Gran,
(1960) and Razavietal. (2015).Vitamin C
was determined by Keitaroet al. (1980)
method.Malondialdehyde(MDA) was
determined by Thiobarbituric acid (TBA)
assays method described by Ohkawaet
al.(1979) and Grotto et al.(2009).
Determination of vitamin E was carried by
method described byRutkowskietal.(2005).
Statistical Analysis
The statistical difference between the levels
of serum Vitamin E, Vitamin C, Calcium and
a malondialdehyde in hypertensive and
normotensive volunteers were determined
using student t test. P<0.05 is considered
significant.
RESULTS
The result of serumVitamin E, Vitamin C,
Calcium and Malondialdehyde (MDA)
levels in hypertensive patients and
normotensive controls attending AKTH are
presented in figures below
The results of this study showed a
significant decrease(p<0.05) of antioxidants
vitamins C and E as well as calcium in
hypertensive patients compared to
normotensive control. However, the results
showed a significant increase(p<0.05) of
serum MDA in hypertensive patients
compared to normotensive controls.

15
a
Serum Calcium(mg/dl)

12

a
9

0
Normotensive Hypertensive

Fig.1: Serum calcium level of hypertensive patients and normotensive controls.Bars with
similar letters are significantly different at (p<0.05).
25
a
a
Serum vitamin C(mg/dl)

20

15

10

0
Normotensive Hypertensive

Fig.2: Serum Vitamin C level of hypertensive patients and normotensive. Bars with similar
letters are significantly different at (p<0.05).
 Oxidative Stress Indices And Calcium

50
a

40
Serum vitamin E (mg/dl)

30

20 a

10

0
Normotensive hypertensive

Fig.3: Serum vitamin Elevel of hypertensive patients and normotensive controls.Bars with
similar letters are significantly different at (p<0.05).

5.00E-05
a

4.00E-05

3.00E-05
Serum MDA(mol/l)

2.00E-05

a
1.00E-05

0.00E+00
Hypertensive Normotensive
Fig.4: Serummalondialdehyde(MDA)level of hypertensive patients and normotensive control.
Bars with similar letters are significantly different at (p<0.05).
.
DISCUSSION (Ayyub et al., 2003). Increased oxidative
The values of calcium, MDA, vitamin C and stress is thought to play a role in the
vitamin E were determined in the serum of development of hypertension complications
both hypertensive and normotensive as in myocardial infarction, stroke and
controls. Hypertension is characterized by chronic kidney disease (Ayyub et al., 2003).
elevated oxidative stress via increased Oxidative stress (OS) and cardiovascular
generation of reactive oxygen species (CV) reactivity are related to CV morbidity
(ROS), and decline in antioxidant defences and mortality.
 Babandi et al., (2017) BJMLS, 2(1):
Antioxidant vitamins E and C are thought to
be effective in increasing the activities of
antioxidant defence enzymes, scavenging
free radicals, preventing oxidative damage
and thereby sparing lipid components of the
cells against lipid peroxidation. OS is
suggested to be a potential contributor to the
development of hypertension and the
associated complications (Zinggetal., 2000).
This may be connected to the fact that the
antioxidant status may be inadequate in
hypertension patients as a result of
overutilization of the antioxidants vitamins
in neutralizing the lethal effectsof ROS in
the system. The metabolic significance of
the evaluation of antioxidants in
hypertension is therefore of paramount
importance.
The decreased levels of antioxidant vitamins
observed in this studymay be connected to
increased oxidative stress in hypertension
resulting in higher utilization of these
vitamins and consequently leading to their
decrease level. Thus, increased intake of
synthetic or natural antioxidant vitamins
could help to avert hypertension
complications (Bunn, 1997). Non-enzymatic
antioxidants are represented byascorbic acid
(Vitamin C), α-tocopherol (Vitamin E) and
other antioxidants. Under normal conditions,
there is a balance between both the activities
and the intracellular levels of these
antioxidants. This balance is essential for the
survival of organisms and their health
(Cadenas, 1997). Non enzymatic
antioxidants such as vitamin C and vitamin
E play an excellent role in protecting the
cells from oxidative damage (Farombiet al.,
2000). Previous findings showed that
vitamin C level is lower in hypertensive
patients compared to general population
(Bates et al., 1998). Khawetal. (2001)
reported a significant association between
plasma vitamin C levels and long term
sequels of hypertension. The finding of this
study showed that vitamin C level
significantly decreased in hypertensive
patients and probably can be implicated in
elevated blood pressure in these patients. It
has been demonstrated that vitamin C
supplementation showed a significant
decline in both systolic and diastolic blood
pressure which may persist for prolonged
period (Khawetal.,2001). In addition,
vitamin C has been suggested to act more
than an antioxidant and its effects on
neurotransmitters lead to its antihypertensive
activity (Hernandez-Guerra, 2006).
Vitamin E level in hypertensive patients was
significantly lower than that of normotensive
volunteers. Vitamin E is a component of the
total peroxylradical-trapping antioxidant
system, reacts directly with peroxyl and
superoxide radicals and singlet oxygen and
protects membranes from lipid peroxidation
(Bisht and Sosodia, 2010). A study by
Bernado Rodriguez-Iturbeetal. (2003)
demonstrated that an antioxidant-enriched
diet that included vitamin E, vitamin C,
selenium and zinc reduces the renal
interstitial inflammation, decreases renal
tissue content of malondialdehyde and
improves management of hypertension.
Furthermore, the cardio-protective potential
of vitamin E has been attributed to its potent
antioxidant action (Choi, 2008). This
contention is supported by the fact that α-
tocopherol shows antioxidant potential by
donating hydrogen radical to remove the free
radicals reacting with it to form non-radical
products or trapping of lipid radicals (Choi,
2008). Vitamin E supplementation will
produce a beneficial effect in hypertensive
patients.
Theobserved increase in Thiobarbituric acid-
Malondialdehyde (TBARS-MDA) level in
hypertensive patients suggest that active
lipid peroxidation is occurring in these
patients. This is in agreement with various
findings of Dhananjayet al. (2013);
Nwanjoet al. (2007) and Ahmad et al.
(2013). Malondialdehyde is a highly toxic
by-product, produced in part by oxidation;
derived from free radicals.
Hypertension is a multi-factorial disorder in
which various physiological mechanisms
participate to elevate BP (Giasuddinet al.,
2001).
 Babandi et al., (2017) BJMLS, 2(1):
Many hypotheses were proposed about the
possible mechanisms underlying essential
hypertension including derangements in
serum electrolytes and water balance. One of
the physiologically important ions in the
serum is calcium. The present study showed
significant decrease in serum calcium in
hypertensive patientscompared with
normotensive volunteers, which is consistent
with the findings of Koschetal (2011).
Calcium is known to play a significant role
in muscle contraction, when calcium is low
then the contraction and relaxation of the
heart is impaired resulting of increase in
peripheral vascular resistance which is
associated with hypertension. Also the
results are consistent with that of Fuet al.
(2001) and Touyzet al. (2000), who reported
a significant decrease in serum calcium in
patients’ with essential hypertension
compared with normotensive volunteers.
Reichelet al. (2001) also reported reduced
calcium in patients with elevated diastolic
blood pressure (DBP). A reduction in
calcium in the diet may cause calcium
depletion from all membrane storage sites,
resulting in less stability of the vascular
smooth muscle cell membrane (Resnick,
1991). When present in optimal
concentrations, calcium stabilizes vascular
cell membranes, inhibits its own entry into
cells, and reduces vasoconstriction (Undurti,
2001). Calcium can combine with some
irons such as magnesium, sodium and
potassium creating ironic balance for
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