J. Afr. Ass. Physiol. Sci.

8 (2): 111-124, December 2020

Journal of African Association of Physiological Sciences

Official Publication of the African Association of Physiological Sciences
http://www.jaaps.aapsnet.org

Research Article
Electrolyte and oxidative stress profile of healthy adult population
in Zaria, Nigeria, and their relationship with experimental pain
response

A.H. Umar1, A. Mohammed1, J.O. Ayo2, N.M. Danjuma3, A.S. Isa1, I. Suleiman1, M.S.
Muhammad4, U.A. Muhammad1, A. Muhammad1, and Y. Yusha’u1.
1
Departments of Human Physiology, Faculty of Basic Medical Sciences, Ahmadu Bello University, Zaria
2
Veterinary Physiology, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria, 3Pharmacology and
Therapeutics, Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Nigeria and 4Human
Physiology, Faculty of Basic Medical Sciences, Gombe State University, Gombe, Nigeria
Keywords: ABSTRACT
Electrolytes, oxidative Background: Electrolyte imbalance and oxidative stress (OS) are known to impair
stress, pressure pain, sex, physiological functions, which can alter health and wellbeing. The reactive species produced
age, ethnicity due to OS are detoxified by endogenous antioxidants to maintain homeostasis. This study
investigated the electrolyte and oxidative stress profile of a healthy adult population in Zaria,
Nigeria and their relationship with experimental pain outcome. Method: Participants were
apparently healthy adult volunteers between the ages of 20 to 65 years and drawn from the city
of Zaria and its environs. Experimental pain was induced using pressure algometry. About 5 ml
of blood was collected for determination of serum electrolytes, malondialdehyde (MDA),
reduced glutathione (GSH) and superoxide dismutase (SOD). Result: The results showed that
serum concentrations of sodium, potassium and chloride as well as oxidative stress profile did
not vary with sex, age and ethnicity among the studied population. There was a significant
negative correlation between pressure pain threshold and serum concentration of potassium (r =
0.2330, p = 0.003) and chloride (r = 0.2126, p = 0.007), while serum sodium correlated positively
(r = 0.3439, p = 0.000). Serum MDA, SOD and GSH did not show statistically significant
correlation with pressure pain threshold (p > 0.05). Conclusion: Serum electrolytes, but not
oxidative stress markers, correlate significantly with experimental pressure pain threshold
among healthy adult population in Zaria, Nigeria.
© Copyright 2020 African Association of Physiological Sciences -ISSN: 2315-9987. All rights reserved

INTRODUCTION biologically important elements play a significant role in

Sodium, potassium, calcium and magnesium are important myriads of physiological functions (Soetan et al., 2010).
elements responsible for electrolyte balance, and their optimal Thus, altered levels of these elements may induce series
concentration for proper biochemical and physiological of events such as slow movement, postural abnormality,
activities of the cell is maintained by ion channels. Potassium,
impaired balance, extensive membrane damage and
sodium, and calcium play integral roles in the
electrophysiology of pain and myriad cellular functions peripheral vascular resistance (Dutta et al., 2015). The
(Kianifard and Chopra, 2018). The optimal concentration of well-being of living cells depends on salinity of
elements responsible for electrolyte balance and proper extracellular fluids. The osmoregulatory system, by
biochemical and physiological activities of living cells is controlling water intake, normally keeps the plasma
maintained by ion channels (Dutta et al., 2015), inactivation sodium concentration within its normal range (135 to
of which lead to altered homeostasis. These 142 mmol per liter), failure of which exposes cells to
hypotonic or hypertonic stress (Sterns, 2015).
Sodium, the most abundant cation in the body, plays a
significant role in fluid balance, osmotic regulation, and
*Address for correspondence: maintenance of membrane potential. Changes in
Email: [email protected] glomerular filtration rate (GFR) can affect normal body
Tel, +2347039271815 sodium concentration. Renal blood flow, prostaglandins,
 Electrolyte and oxidative stress profile in experimental pain response

and natriuretic peptides also play a role in sodium Increase in oxidative stress causes disturbance of
regulation, with the Na+-K+-ATPase pump maintaining homeostasis, leading to denaturation of intracellular
cellular membrane potential. Sodium intake, under molecules like nucleic acids, proteins, and lipids by ROS
normal physiologic condition, matches sodium losses. (Chang et al., 2013; Rubolini et al., 2012). Indeed, ROS
Abnormally high sodium concentration (hypernatremia) can cause different degrees of oxidative damage to
presents with increased thirst, fatigue, restlessness, and biological macromolecules and cells/ tissues. Although
muscle irritability, and in severe conditions, cerebral some ROS play a key role in cell signalling, oxidative
cellular dehydration can occur, which can progress to stress is known to impair physiological functions, may
hemorrhage, seizures, coma, and death (Blackmer, accelerate ageing, and cause higher susceptibility to
2018). Hyponatremia, on the other hand, is associated environmental stress or pathogens, thereby resulting in
with headache, nausea, myopathy, lethargy, and reduced fitness (Rubolini et al., 2012). Oxidative stress
restlessness and seizures. Potassium, the primary causes destruction of cell membrane and cartilage,
intracellular cation, also play essential roles in cellular breakage of DNA strand, rise in intracellular free Ca2+
metabolism and maintains membrane potential as well and damage to membrane ion transporters, thus, leading
as promotes neuromuscular and cardiac function to cell death and disease development (Dutta et al.,
(Blackmer, 2018). 2015). Decrease in CAT and/or SOD and reduced levels
of free radical scavengers such as glutathione, and
Oxidative stress is the imbalance between the production vitamins C and E also contribute to oxidative stress
of pro-oxidant reactive species and the ability of the (Baradaran et al., 2014).
living system to cope and prevent their adverse effects
via antioxidant defence. It is the persistent imbalance Oxidative stress has been implicated in perception of
between the production of Reactive oxygen species pain. For example, NO is involved in perception and
(ROS) and antioxidant defences, and it often results in reduction of pain caused by osteoarthritis (OA) via
irreversible cellular damage (Prasad et al., 2017). ROS normalization of blood flow pathway (which may help
are oxygen containing molecules including free radicals to decrease ischemic pain), the nerve transmission
(hydroxyl radical, superoxide anion, hypochlorite ion), pathway (which decreases the irritation of the nerves in
hydrogen peroxide, and nitric oxide. Due to the presence the synovium, bone, and soft tissues), the opioid receptor
of unpaired electrons in the valence shell, free radicals pathway (which might stimulate the body's normal pain
are short-lived, unstable and highly reactive. ROS are reduction pathways), and the anti-inflammation pathway
generated by the mitochondria (through oxidative (Lepetsos and Papavassiliou, 2016). Both H2O2 and
phosphorylation), non-mitochondrial membrane-bound ONOO− are involved in inflammatory pain, mainly
nicotinamide adenine dinucleotide phosphate (NADPH) through COX2/PGE2 pathway.
oxidase, and xanthine oxidase (XO) (Lepetsos and
Papavassiliou, 2016). ROS are formed as byproduct of The ROS generated during normal cellular metabolism
normal metabolism. These molecules play an important can be scavenged by enzymatic antioxidants (SOD, GPx,
role in cell signalling pathways and likely act as key CAT), or non-enzymatic antioxidants (mainly vitamin E
players in many disorders (e.g. metabolic syndromes) and reduced form of Glutathione). However, oxidative
and homeostasis. They include oxygen radicals and stress results either due to increased ROS production or
reactive non-radicals (Banerjee et al., 2016). Nitric oxide decreased antioxidant defense systems or both (Prasad et
(NO), a signaling molecule in the Central Nervous al., 2017). The brain contains high concentration of poly-
System (CNS), reacts with excess ROS to generate unsaturated fatty acids that are highly susceptible to lipid
Reactive Nitrogen Species (RNS) like peroxynitrite, peroxidation. Thus, assessment of MDA levels in
which is also a powerful oxidizing agent that can cause biological materials can be used as an important
lipid peroxidation, thus contributing to free radical- indicator of lipid peroxidation in vitro and in vivo for
mediated damage (Prasad et al., 2017). The ROS are various diseases
detoxified by endogenous anti-oxidant system of
enzymatic and non-enzymatic antioxidants such as Free radicals attack polyunsaturated fatty acid of the
superoxide dismutase (SOD), catalase (CAT), and membrane, leading to lipid peroxidation and excessive
glutathione peroxidase (GPx), glutathione (GSH), production of malondialdehyde (MDA), the most
NADPH ubiquinone oxidoreductase (NQO1), abundant product of lipid peroxidation. MDA reacts
paraoxonases (PON), ascorbic acid (vitamin C), α- with membrane proteins and ion channels, affecting their
tocopherol (vitamin E) and carotinoids, which help to normal function and causing electrolyte imbalance
keep the system in a state of normal homeostasis. (Dutta et al., 2015). This study aimed to investigate the

112 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Umar et al.

 Electrolyte and oxidative stress profile in experimental pain response

hypothesis that serum electrolytes and oxidative stress preparation reagent (sodium RI), 10 µL of standard and
does not correlate with experimental pain outcome 10 µL of serum were vigorously mixed and incubated at
among healthy adult population in Zaria, Nigeria. room temperature for 5 minutes, followed by
centrifugation at 1000 g to obtain a clear supernatant.
MATERIALS AND METHODS Supernatant was transferred for standard and test, and
Subject recruitment the assay and absorbance was measured at 530 nm.
This research was carried out in nine randomly selected Serum potassium was determined by colourimetric
secondary schools under the Zaria Zonal Education method of Tietz (1976). A volume of 1000 µL of
Authority. Observational cross-sectional study was used reagent, 25 µL of standard and 25 µL of serum were
for this research. One hundred and sixty one (161) mixed and incubated at room temperature for 5 minutes.
apparently healthy volunteers of ≥ 20 years were The mixture was then centrifuged at 1000 g and the clear
recruited for the study. The subjects included mostly supernatant transferred for standard and test assay.
secondary schools staff population around Zaria and Absorbance was measured at 530 nm against distilled
environs in Kaduna State, Northern Nigeria. Sample water. Determination of serum cloride concentration
recruitment was based on convenience, agreeing with was carried out by colourimetry method of Schonfeld
the study protocol, qualified and willing to participate. and Lowellen (1964). Volume of 1000 µL of reagent, 10
Selected individuals were taught and given basic µL of standard and 10 µL of serum were thoroughly
understanding about Physiology of pain and were mixed and allowed to incubate at room temperature for
informed about the experimental procedures (verbally 1 minute. Absorbance of standard and test samples was
and in writing), as well as risks and contraindications of measured at 530 nm against distilled water.
all procedures. Informed consent was obtained from the
subjects, as well as ethical approval from the Ahmadu
Bello University Teaching Hospital Health Research Determination of oxidative stress
Ethics Committee (ABUTH/HREC/N22/2015) prior to Serum MDA was measured using the method of Buege
the study. Subjects were excluded if they reported and Aust, (1978), as described by Dsouza et al. (2012).
significant psychiatric co-morbidity, prior or present 100 μL of serum was diluted to 500 μL with distilled
alcohol abuse, use daily analgesics or have any water, and the samples were kept in boiling water bath
neurological or inflammatory disease that could interfere for 15 min. 1 mL of Trichloroacetic acid (TCA)-2-
with pain perception and pain report, such as diabetes, thiobarbituric acid (TBA)–HCl reagent was added and
peripheral or central neuropathy, a chronic pain disorder, the reaction mixture was cooled to room temperature,
or current pain condition. Patients with a diagnosis of and centrifuged at 1000 g for 5 min. The supernatant was
dementia were also excluded. taken and the optical density of the pink colour formed
was read in a spectrophotometer at 535 nm. Serum SOD
Experimental pressure pain assessment activity was determined by its ability to inhibit auto-
Pressure pain was assessed by applying pressure at the oxidation of epinephrine which was estimated by the
first dorsal interosseous muscle (Chesterton et al., 2007), increase in absorbance at 480 nm, as described by Sun
using pressure algometer, at the rate of 1Kg/sec. and Zigma (1978). Enzyme activity was calculated by
Subjects were told to press a button at the first perception measuring the change in absorbance at 480 nm for 5
of pain. The pressure at which pain was first reported minutes. Serum GSH concentration was estimated
was recorded as the pressure pain threshold (in Kg). according to the method described by Sedlak and
Retest was not carried out as study has demonstrated Lindsay (1968) and Awoyemi et al. (2014). To 0.5ml of
good test–retest reliability for repeated measurements serum, 2.5ml of 0.3M phosphate buffer was added,
(Lacourt et al., 2012). Also, pressure pain was recorded followed by 0.5 ml of Ellman’s reagent. The mixture was
on the dominant hand only, as reports indicated no incubated for 10 minutes and the absorbance was read at
significant difference between dominant and non- 412 nm using spectrophotometer.
dominant sides (Fischer, 1987; Park et al., 2011).
Hogeweg et al. (1992) and Rui et al. (2015) also reported
no significant differences in pressure pain thresholds Statistical Analysis
between the same points on either side of the body. All analyses were carried out using SPSS version 23
software for windows (SPSS Inc, Chicago). Results are
Determination of serum electrolytes: presented as mean ± SEM. Sex differences were
Serum sodium was determined by colourimetry method analysed by Independent-Samples t test, while age and
of Maruna and Trinder (1958). A volume of 1000 µL of ethnic differences were analysed by analysis of variance

113 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Umar et al.

 Electrolyte and oxidative stress profile in experimental pain response

(ANOVA), followed by Tukey’s post hoc tests to statistically significant [p=0.527; T(155)=-0.634].
indicate significance (if any). Associations were Difference in serum potassium concentration also
determined using Pearson’s correlation. Values of p < showed no statistical sex significance [p=0.099;
0.05 were statistically significant. T(155)=-1.660], but male participants have lower
concentration (5.09±2.08), than females (5.77±3.09).
RESULTS Analysis of serum chloride concentration shows that
Sociodemographic characteristics male participants had slightly lower serum chloride
Out of the 161 participants included in the study, 91 are (93.42±11.89) than females (94.10±10.13), but the
males (56.5%), while 70 are females (43.5%). Age difference was not statistically significant [p=0.706;
distribution showed that 36% are between 20 to 30 years, T(155)=-0.378] (table 3). The result of age differences
31.1% between 31 to 40 years, 16.8% are between 41 to in serum electrolyte concentration showed that serum
50 years, while 14.9% are above 50 years. By ethnicity, sodium concentration was significantly higher among
60.9% belong to Hausa ethnic group, 15.5% Yorubas, subjects in the younger age group of 20 – 30 years
8.1% Fulanis, and the other minority groups, classified (165.06±50.11), than those in the 41 – 50 years group
as Others, constituted 13.7% (119.54±44.61) (p=0.001) and those in the above 50
years group (129.76±11.73) (p=0.018). Age difference
Pressure pain in serum potassium concentration shows no statistically
This study was carried out on 161 apparently healthy significant variation among the studied subjects
volunteers, made of 70 (43.5%) females (table 1). (p=0.076; f=2.342), though there was a decrease with
Perception of pressure pain was found to be significantly increase in age, as participants in the youngest age group
lower in males than in females. Mean pressure pain (20 – 30 years) had the highest concentration
threshold was significantly higher in males (6.90±0.14) (6.03±2.91), and those in the oldest age group (above 50
compared to females (6.00±0.17) (p = 0.00; T = 4.095) years) had the lowest concentration (4.45±2.13). Serum
(figure 1). Perception of pressure pain in the present chloride concentration analysis shows no statistically
study showed that subjects in the 41 – 50 age group had significant age variation among the studied participants
significantly lower pressure pain threshold (5.85±0.24) (p=0.361; f=1.076), but the result shows that there is an
when compared to all the other groups (ANOVA; p < age dependent increase, with participants in the youngest
0.05; F = 2.221). The result also showed that participants age group having the lowest concentration
that fall in the youngest age group (20 to 30 years) had (92.25±10.29), and those in the oldest age group having
the highest pressure pain threshold (6.67±0.19), while the highest (96.04±13.32) (table 4)
those in the oldest age group (>50) had a threshold of
6.61(0.29), which is lower than the youngest age group.
Table 1: Socio-Demographic Characteristics of Study
Participants in the 31 to 40 years group have a threshold
Volunteers, Showing Frequency and Percentage
of 6.63(0.21) (figure 2). Ethnicity was found to affect
Distribution for Sex, Age and Ethnicity in an Adult
perception of pressure pain among healthy adult
Population of Zaria, Nigeria.
population in Zaria, Nigeria, in the present study, with
Yorubas having statistically significant higher pressure
pain threshold (7.32±0.30) compared to Hausas Characteristic Description Frequency Percentage
(6.28±0.12) (p = 0.008; f = 4.143). Participants Sex Male 91 56.5
belonging to Fulani ethnic group had a threshold of Female 70 43.5
6.17(0.43), showing that they have a higher pressure Total 161 100
pain perception than the Hausas and Yorubas, though the
Age (years) 20 – 30 58 36.0
differences were not statistically significant. Others
ethnic group showed a mean threshold of 6.83(0.40), 31 – 40 50 31.1
which is higher than Hausas and Fulanis, but lower than 41 – 50 27 16.8
the Yorubas, though all the differences showed no >50 24 14.9
statistical significance (figure 3). Total 159 98.8
Ethnicity Hausa 98 60.9
Serum electrolytes
The result of sex differences in serum electrolytes Yoruba 25 15.5
showed that the male subjects had lower serum sodium Fulani 13 8.1
concentration (143.65±52.95) than the females Others 22 13.7
(148.83±48.71), though the difference was not Total 158 98.1
114 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Umar et al.
 Electrolyte and oxidative stress profile in experimental pain response

Table 2: Age differences in mean anthropometric characteristics in a healthy adult Nigerian population in Zaria

Characteristic/ 20 – 30 31 – 40 41 – 50 Above 50
Age (yrs) (Mean±SD) (Mean±SD) (Mean±SD) (Mean±SD)
b
Weight (Kg) 61.29±10.42 71.02±14.06 73.16±10.19 76.68±15.39
Height (m) 1.65±0.09 1.67±0.08 1.66±0.09 1.64±0.08
BMI (Kg/m2) 22.46±3.47a 25.44±4.64b 26.67±3.98b,c 28.40±5.38c
SBP (mmHg) 107.58±14.55a 114.79±9.84a,b 122.96±26.28b,c 133.54±15.07c
DBP (mmHg) 72.15±10.47a 76.26±7.61a,b 79.63±16.46b 88.12±11.87c
Mean difference of values with different superscript letters ( a,b,c) is statistically significant (p < 0.05) across the table (ANOVA,
df = 157). SBP (systolic blood pressure); DBP (diastolic blood pressure).

Table 3: Sex Differences in Serum Electrolytes and Oxidative Stress Markers Among Healthy Adult Population in
Zaria, Nigeria
Parameters Male Female
[Na] (mmol/L) 142.15±5.75 148.83±5.95
[K] (mmol/L) 5.19±0.24 5.99±0.40
[Cl] (mmol/L) 93.42±1.25 94.10±1.23
MDA (nmol/ml) 23.68±1.53 23.15±1.31
GSH (µg/ml) 30.68±0.68 21.79±0.78
SOD (µg/ml) 13.51±1.01 14.07±0.98

Fig. 2: Age Variation in Experimental Pressure Pain

Fig. 1: Sex Differences in Experimental Pressure Pain Threshold in a Healthy Adult Population in Zaria, Nigeria,
Threshold in a Healthy Adult Population of Zaria, Nigeria. Showing Age Differences in Pressure Pain Threshold. a,bMean
a,b
mean difference is statistically significant between males difference is statistically significant between groups
and females (independent t-test; p = 0.00); (n). (ANOVA; p < 0.05); (n).

115 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Umar et al.

 Electrolyte and oxidative stress profile in experimental pain response

Nigeria. *Mean difference between the groups is statistically

significant (ANOVA; p < 0.01) ; (n).
Result of ethnic differences in serum concentration of
sodium, potassium and chloride showed that subjects
that belong to the Hausa tribe had statistically significant
lower serum sodium (135.21±47.37) than the Yorubas
(187.94±35.00) (p=0.000; f=11.054) and those in the
others group (167.27±48.83) (p=0.023). The Yorubas
had significantly higher serum sodium than the Fulanis
(123.22±51.15) (p=0.001), and the difference between
the Fulanis and Others was also statistically significant
(p=0.045). Ethnic difference in serum potassium
concentration showed no statistically significant
variation among the studied subjects (p=0.063; f=2.486),
though the Fulanis had the highest concentration
(6.26±3.25), while those in the Others group had the
least (4.20±2.03). Serum chloride concentration analysis
Fig. 3: Comparison of Pressure Pain Threshold between showed no statistically significant ethnic differences
Ethnic Groups among a Healthy Adult Population in Zaria, among the studied population (p=0.193; f=3.148), but
the result showed that subjects in the Fulani group

Table 4: Age Differences in Serum Electrolytes and Oxidative Stress Markers Among Healthy Adult Population in Zaria, Nigeria
Age (years)
Parameters 20-30 31-40 41-50 > 50
[Na] (mmol/L) 165.06±6.69a 143.79±6.59a,b 119.54±8.75b 129.76±11.73b
[K] (mmol/L) 6.04±0.39 5.28±0.33 5.09±0.49 4.45±0.44
[Cl] (mmol/L) 92.25±1.37 93.08±1.71 96.04±1.79 96.04±2.78
MDA (nmol/ml) 24.47±2.19 22.33±1.02 22.48±1.30 23.59±1.39
GSH (µg/ml) 29.88±0.84 31.50±0.85 28.04±1.43 31.33±1.27
SOD (µg/ml) 15.68±1.54 12.76±0.52 11.24±0.69 12.83±0.63
a,b,
Mean differences of values with different superscript letters are statistically significant along the column
(ANOVA, p < 0.05)

Table 5: Ethnic Differences in Serum Electrolytes and Oxidative Stress Markers Among Healthy Adult Population
in Zaria, Nigeria
Ethnicity
Parameters Hausa Yoruba Fulani Others
[Na] (mmol/L) 133.89±4.99a 187.94±7.00b 123.22±14.77a 167.27±10.65b
[K] (mmol/L) 5.63±0.27 4.96±0.41 6.26±0.94 4.20±0.44
[Cl] (mmol/L) 95.22±1.05 89.40±2.63 95.92±2.87 89.52±2.32
MDA (nmol/ml) 22.76±0.85 26.25±3.49 22.84±1.78 21.66±1.27
GSH (µg/ml) 30.58±0.77 30.12±1.07 28.74±2.13 30.44±1.05
SOD (µg/ml) 13.05±0.53 16.30±2.44 10.71±0.76 13.73±0.96
Mean differences of values with different superscript letters a,b, are statistically significant along the column (ANOVA, p < 0.05)

had the highest (95.92±9.94), and the Yorubas had the (23.68±1.53) than the females (23.16±1.31), though the
lowest (89.40±13.14) (table 5) difference was not statistically significant [p=0.762;
T(155)=0.259]. Difference in serum GSH concentration
Oxidative stress does not also statistically vary by sex [p=0.391;
Sex differences in serum concentration of MDA, GSH T(155)=0.865], but the males had higher GSH
and SOD in the present study showed that the male concentration (30.69±0.68) than the females
subjects had slightly higher serum MDA concentration (29.79±0.78). Analysis of serum SOD
(23.68±1.53) than the females (23.16±1.31), though the
116 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Umar et al.
 Electrolyte and oxidative stress profile in experimental pain response

117 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Umar et al.

 Electrolyte and oxidative stress profile in experimental pain response

concentration showed that male participants had lower DISCUSSION

serum SOD (13.51±1.00) than the females (14.07±0.98), Biologically important elements have significant role in
but the difference was also not statistically significant maintenance of homeostasis and participate in various
[p=0.740; T(155)=-0.402] (table 3). physiological activities such as neuromuscular
irritability, nerve conduction, prevention of
The result of age differences in serum concentration of development of age-elated complications, among others.
MDA, GSH and SOD showed that serum MDA Alteration in the levels of these elements may induce
concentration was highest among subjects in the series of events such as slow movement, postural
youngest age group of 20 – 30 years (24.47±2.19), and abnormality, impaired balance, extensive membrane
lowest among the 31 – 40 years group (22.33±1.02), damage and peripheral vascular resistance (Dutta et al.,
though the differences were not statistically significant 2015). Relationship betwen electrolyte profile and
(p=0.829; f=0.296). Age difference in serum GSH clinical pain has not been well documented and
concentration showed no statistically significant measuring electrolytes in sickle cell aneamia patients
variation among the studied population (p=0.159; presenting with painful vaso-occlusive crises has been
f=1.764). Participants belonging to the 41 – 50 years shown to be un-necessary (Desai, 2014).
group had the lowest serum GSH (28.04±1.43), while
those in the 31 – 40 years group had the highest Although serum electrolytes concentrations did not
(31.50±0.85). Analysis of serum SOD concentration statistically vary by sex in the present study, serum
also showed no statistically significant age variation sodium concentration was found to decrease with age, as
among the studied population (p=0.117; f=2.017), but subjects in the younger age group of 21 – 30 years have
there was a somewhat age dependent decrease, with significantly higher serum sodium than those in the 41 –
participants in the youngest age group (20 – 30) having 50 years group and those in the above 50 years group.
the highest concentration (15.68±1.54), and those in the Our result is in agreement with that of McLean et al.
41 – 50 years group having the lowest concentration (2003), who found no sex difference in serum sodium
(11.24±0.69) (table 4). and potassium concentrations. Singh et al. (2016) also
reported no significant sex differences in serum sodium,
Ethnic differences in serum concentration of MDA, potassium and chloride concentrations. Our result also
GSH and SOD showed that subjects that belong to the agree with a previous work by Prabhunath et al. (2016)
Yoruba ethnic group had the highest serum MDA that reported a significantly lower serum sodium
concentration (26.25±3.49), while those in the Others concentration in healthy elderly subjects than their
group had the least (21.66±1.27), but there were no younger counterparts, but they also reported a
statistically significant variations among all the ethnic significantly higher serum potassium concentration,
groups (p=0.426; f=0.938). Ethnic difference in serum contrary to our finding. Ibrahim et al. (2012) reported no
GSH concentration also showed no statistically- significant sex difference in serum sodium, but
significant variation among the studied population significantly higher serum potassium and chloride in
(p=0.792; f=0.346), though the Fulanis had the lowest female than male Turkeys, as well as higher serum
GSH concentration (28.74±2.13), while the Hausas had sodium, potassium and chloride in adults. Decreased
the highest (30.58±0.77). Serum SOD concentration serum sodium may be as a result of hormonal
analysis showed no statistically significant ethnic imbalances, such as the syndrome of inappropriate ADH
differences (p=0.136; f=1.894). The Fulani ethnic group secretion, a common occurrence in elderly (Prabhunath
had the least serum SOD concentration (10.71±0.76), et al., 2016).
while the Yorubas had the highest (16.30±2.44) (table Potassium and sodium are known to play integral roles
5). in the electrophysiology of cellular functions, including
pain transmission (Kianifard and Chopra, 2018).
There was a significant positive correlation between Lidocaine, for example, is an amide-type local
serum sodium and experimental pressure pain in the anaesthetic that exerts its pharmacological action
present study (p = 0.000) (figure 4), while serum through the block of sodium channels in neural tissues,
potassium and chloride correlated negetively and thereby interrupting neuronal transmission (Eipe et al.,
significantly with experimental pressure pain (p = 0.003; 2016), including blockage of pain transmission. The lack
p = 0.007) (figures 5 and 6). Serum MDA, GSH and of significant effect of sex, age and ethnicity on serum
SOD, on the other hand, showed no signifcant potassium in the present study may explain the outcome
correlation with experimental pressure pain in this study. of cold pressor pain responses, as potassium ion has been
118 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Umar et al.
 Electrolyte and oxidative stress profile in experimental pain response

implicated to play a role in abnormal pain signalling. Sex ageing. Elderly patients are more likely to develop
and age variations in experimental pressure pain extracellular fluid volume depletion due to delay in the
threshold in the present study is in tandem with the ability of the ageing kidneys to lower sodium excretion
observed differences in serum potassium concentration to minimal value (Luckey and Parsa, 2003).
(higher pain threshold is associated with higher serum The somewhat age-dependent decrease in serum sodium
potassium, as seen in the results). The Kv7 (KCNQ or M concentration in this study may be associated with an age
channels) family is widely expressed in the nervous related alteration in blood electrolyte balance. Kengne et
system, including nociceptors. Injection of M channel al. (2008) reported hyponatraemia as a risk of fracture in
blocker in the hind paw of rats induced moderate pain, ambulatory elderly. Older subject’s kidneys have been
while injection of M channel enhancer produced an reported to possess an inability to conserve urinary
analgesic effect. Reduced Kv7 function was also sodium. Decrease in serum aldosterone may account for
involved in inflammatory pain (Kianifard and Chopra, an increase in excretion of sodium in the urine and an
2018). Diclofenac which is popularly used to treat pain inability to decrease urinary sodium excretion in a
and inflammation was demonstrated to activate hypovolemic state. Age induced decrease in glomerular
Kv7.2/Kv7.3 channels. filtration rate (GFR) is associated with a decline in the
Though not significant, the female subjects were found number of functioning glomeruli, and a corresponding
to have higher sodium concentration than the males, reduction in number of active kidney tubules. This leads
which may be due to differences in sex hormone level, to a decrease in the ability to dilute urine and an
as plasma rennin activity has been reported to correlate increased prevalence of hyponatremia in the elderly
positively with serum oestradiol level (Philips et al., (Schlanger et al., 2010). The aged kidney is also
1995). The lack of significant sex differences in serum associated with abnormal distal tubular activity, which
electrolytes in this study may be due to inability to assess leads to a decreased ability to reabsorb sodium. This can
the menstrual status of the female subjects, as both be a consequence of reduced GFR or decreased
serum sodium, potassium and chloride concentrations responsiveness of the tubules to ADH. There is also an
were reported to vary across the various stages of the age related decrease in renin activity with aldosterone
menstrual cycle, with highest concentrations found secretion (Prabhunath et al., 2016).
during the ovulatory phase (Dadlani et al., 1981). Age related changes in serum electrolyte concentrations
Pregnancy and lactation, which were also not assessed may be due to age-related reduction in the serum
in this study, may also influence serum sodium concentrations of renin and aldosterone consequent to
concentration, as Faith et al. (2017) reported increased ANP activity, usually released in response to
significantly higher serum sodium in lactating ewes increased blood pressure and right atrial filling (El-
when compared to pregnant ewes, though Obembe and Sharkawy et al., 2014). The elderly are commonly
Antai (2008) reported no significant difference in known to be prescribed many medications that interfere
concentration of serum electrolytes between pregnant with urinary excretion of electrolytes/potassium
and non-pregnant women. (Schlanger et al., 2010).
Studies indicate an age-related decrease in plasma The significant ethnic difference in serum sodium
sodium level by about 1 mEq/L per decade (Luckey and concentration observed in this study is likely to be
Parsa, 2003). Though subjects in the oldest age group attributed to dietary intake. Yousafzai et al. (2011)
have higher blood pressure (Table 2), they also have reported ethnic differences in serum sodium and chloride
lower serum sodium. This may be associated with between Pathan, Baloch and Punjab patients, a
increase in plasma volume due to decrease in diluting difference they associated to diet. Chun et al. (2008)
capacity of the kidneys, accounted for as a result of reported no significant difference in baseline sodium
increase in solute load delivery to the remaining excretion between blacks and whites, as well as after
functioning glomeruli. The Na-K-2Cl co-transporter, furosemide injection, but there was statistically
which plays dual roles in the kidney’s capacity to significant decrease in potassium excretion in blacks
concentrate and dilute the urine, has been shown to be than in whites. A study by Kim et al. (2017) concluded
down-regulated in aged rodents. Decrease in the that Hispanic haemodialysis patients have higher serum
reabsorption of NaCl increases solute delivery to the potassium levels than African-Americans and whites,
collecting tubule and decreases solute free water while higher death risk is associated with higher
excretion (Schlanger et al., 2010). Alterations in the potassium levels in African-Americans and whites, but
level of aldosterone, antidiuretic hormone (ADH) and not in Hispanics. This was postulated to be associated
atrial natriuretic peptide (ANP) in the elderly are partly with socio-cultural factors such as high potassium
responsible for changes in fluid balance associated with tolerance due to very high potassium diet, or

119 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Umar et al.

 Electrolyte and oxidative stress profile in experimental pain response

racial/ethnic genetic differences in potassium glutathione synthesis depends on the availability of

adaptation. Rate of disposal of intravenous load and cysteines, the rate-limiting precursor, and diurnal
urinary excretion of potassium has been reported to be variation in GSH and cysteine has been reported, and
less in African-Americans when compared with GSH levels and GSH/GSSG ratio has been related to
Caucasian-American subjects, a difference associated pathological conditions (Marocco et al., 2017).
with potassium uptake in skeletal muscle by Na/K
ATPase (Suh et al., 2004). African-Americans were Glutathione (GSH) is the most abundant thiol
reported to have lower serum potassium values than antioxidant present in the blood and tissues, together
non–African Americans independent of demographic with its associated biosynthetic, redox and detoxification
characteristics, comorbid conditions, and potassium- pathways represent the key defense system against
altering medications. African ancestry was associated oxidative stress and free radical damage in the cell
with lower serum potassium levels, showing a genetic (Thomas et al., 2015). Though glutamate is significantly
component to potassium homeostasis (Chen et al., lower in males than females in the present study, GSH
2017). The African ancestry of the subjects in the present was found to be opposite, though not significant. The
study could explain the lack of statistically significant lack of significant difference in serum GSH may be
age and ethnic differences in the measured serum similarity in diet among the studied population.
electrolytes. The present study agreed with the finding of Sobocanec
et al. (2009) that reported no significant sex or age
The present study showed no statistically significant sex differences in brain SOD activity between male and
variation in serum MDA, GSH and SOD levels. This female mice, but disagree with Pepe et al. (2009) that
agrees with previous findings that reported no significant reported significantly higher resting SOD activities in
sex difference in serum MDA level (Beg et al., 2005; El- men than women. SOD is an enzyme that is known to
Badry 2006; Moreto et al., 2014). Schmitt et al. (2016) provide protection against free radical induced damage
also reported no significant sex difference in serum by converting superoxide radicals (O2−) generated in
MDA in normal control rats. However, this study peroxisomes and mitochondria to hydrogen peroxides.
disagrees with the work of Tothova et al. (2012), who The hydrogen peroxide is then converted to water and
reported significantly lower MDA in males than molecular oxygen by catalase (CAT). Inhibition of SOD
females, while Bhutia et al. (2011) reported higher leads to increased oxidative stress as a result of the
serum MDA levels in male than female diabetics. Lack damageing activities of the superoxide free radicals, and
of sex difference in serum MDA in the present study also reduction in the activity of CAT (Awoyemi et al.,
may be associated with menstrual status of the female 2014). Sex differences in antioxidant enzymes may be
subjects, as MDA level has been reported to be attributed to factors such as age, sex hormones,
significantly higher during the luteal phase when cardiovascular health, genetics and lifestyle (Pepe et al.,
compared to the follicular and ovulatory phases (Akande 2009).
and Akinyinka, 2005). Age difference in serum markers of oxidative stress
shows no statistically significant variation in the present
Sex differences in oxidative stress markers and study. Though ageing has been associated with increased
enzymatic antioxidant defence mechanisms are oxidative stress supposedly due to increase free radical,
attributed to higher levels of sex hormones; oestradiol the lack of significant age difference in the present study
and other oestrogens in fertile females (Tothova et al., may be due to small number of aged participants, as only
2012). Trevisan et al. (2001) reported lower glutathione 24 subjects (14.9%) were above 50 years. The present
level in men than in women, while Schmitt et al. (2016) study agrees with that of Bogdanska et al. (2003), but
reported no sex difference in erythrocyte GSH in control disagrees with the finding of Mecocci et al. (2000) that
rats. GSH, a tripeptide representing the most abundant reported increase of both plasma and red blood cell SOD
nonprotein thiol present in the cell, acts as an antioxidant activity with age. Guemeun et al. (1991) reported no
defence system by its ability to scavenge reactive statistically significant variation of plasma and
oxygen species (ROS) through the reversible oxidation erythrocyte SOD activities related to sex, but plasma and
to oxidized glutathione (GSSG) (Frijhoff et al., 2015). erythrocyte SOD correlate negatively with age. Ho et al.
Lack of significant difference in serum GSH in the (2005) reported a significant increase in SOD activities
present study may be associated with the fact that with age, but no significant sex variation.
glutathione is minimally concentrated in serum as it is The present study showed no significant effect of
mainly stored within the cytosol. Diet, diurnal variation ethnicity on oxidative stress biomarkers. A study by
and health status affect GSH concentration, as Bhutia et al. (2011) showed that serum MDA levels of

120 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Umar et al.

 Electrolyte and oxidative stress profile in experimental pain response

type 2 diabetics in different ethnic populations of Sikkim injuring ion channels, including Na+-K+ ATPase, Na–
showed no significant difference. Lipid peroxidation Ka–2Cl co-transporter and K+ channels, as well as
impairs nerve tissue structure and function by decreasing production of protein radical in lipid membranes that
the activity of Na/K ATPase. Painful procedures are well effects normal ion transport (Dutta et al., 2015). This is
recognized conditions leading to free radicals clearly demonstrated by low level of trace elements and
generation, and oxidative stress plays a role in pain electrolytes and high oxidative stress index in camel-
mechanisms, especially in neuropathic and calves with pasteurellosis (Shoieb et al., 2016).
inflammatory pain. Antioxidants have also been used as Limitations of the present study include inability to
anti-nociceptive agents. Reactive oxygen species play an determine the pregnancy and menstrual status of the
important role in allodynia and hyperalgesia mainly female volunteers, as well as the diet of the studied
through central sensitization, as well as in the population.
pathogenesis of rheumatoid arthritis (Szallasi et al.,
2011). CONCLUSION
The lack of significant difference in oxidative stress The present study was able to establish that sex
markers in this study may be due to the fact that healthy differences does not determine the concentration of
subjects with no apparent medical condition were used, serum electrolytes among the studied population, but
and oxidative stress is a pathophysiologic imbalance serum sodium decreases with age and is also affected by
between oxidants and antioxidants system (Chang et al., ethnicity, most likely due to difference in dietary intake.
2014), which result from increased production of free Serum MDA, SOD and GSH showed no variation by
radicals and/or decreased antioxidant defence sex, age or ethnicity. Pressure pain threshold correlates
mechanisms, and it plays a significant role in several positively with serum sodium concentration, but
pathological conditions and their complications, such as negatively with serum potassium and chloride
cardiovascular diseases, cancer, inflammation, and concentrations.
chronic stress. During pain processing, there is
involvement of reactive species including superoxide, ACKNOWLEDGEMENT
NO, and peroxynitrite, and increase in SOD activity is We acknowledge the contributions of Mal. Ya’u Bello
associated with decreased availability of superoxide and Mal. Bala Muhammad of the Human Physiology
(Kolberg et al., 2015). laboratory, for helping with logistics and transport
Lipid peroxidation has a wide range and depends upon during the period of the research.
several factors like diet and lifestyle of the individual
(Beg et al., 2005). Lipid peroxidation caused by ROS CONFLICT OF INTEREST
causes disarrangement and disruption of cell The authors hereby declare that there is no conflict of
membranes, leading to necrotic cell death. It is a free interest regarding the publication of this paper.
radical–related process that is associated mostly with
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