40 Journal of Jazan University - Applied Sciences Branch
Vol.3 No.1 Jan. 2014 (Safar 1435 H)
Serum Protein Electrophoretic Pattern Among Khat Chewers
Serum Protein Electrophoretic Pattern
Among Khat Chewers
M.M. Halawi1, H. I. AbuTaweel1, R. B. M. Al-Sanousi2 and S. I. Abdelwahab2*
1
Medical Research Centre, Jazan University, KSA
Substance Abuse Research Centre, Jazan University, KSA
2
Abstract
The leaves of khat (Catha edulis) are found to have stimulating and pleasurable
effects and are chewed habitually by people of East Africa and some Arabian
Peninsula, Yemen and Southern kingdom of Saudi Arabia. Due to the habitual
widespread use of khat in Jazan area the present study was undertaken to evaluate the effects of khat on the protein electrophoresis pattern in khat chewing
males. A total of 70 subjects in which 50 were khat users and 20 were non-khat
users were subjected to this study. Serum total proteins,albumin and α1, α 2, β
and γ globulins were analysed. The results of the present study showed that serum total proteins, serum albumin and α1 were elevated in 16.66%, 33.7% and
31.5% of all cases respectively, while γ globulin was decreased in 35% of studied cases. Disturbances in the protein levels and electrophoretic pattern are new
addition side effects due to khat chewing as far as we know this is the first paper
in this field.
Keywords: Khat chewing; protein electrophoresis; electrophoretic pattern
1.INTRODUCTION
Khat (Catha edulis, Celestraceae) is a
flowering plant, indigenous to tropical
East Africa and the Arabian Peninsula.
The origins of the plant are often argued.
Many believe its origins are Ethiopian,
others state that khat originated in Yemen
before spreading to Ethiopia and the nearby countries (Balint et al., 2009; Vekariya,
2012). Khat grows in habitats varying
from evergreen submontane forest to deciduous woodland at 800–2000 m altitude
and is now indigenous in Ethiopia, Kenya,
Uganda and Tanzania, and from East
Congo (formerly Zaire) southward to
South Africa (Balint et al., 2009; Balint
and Balint, 1994). Chewing the leaves of
the plant for their pleasurable stimulant effect is a habit that is widespread in the
aforementioned geographical areas. It is
estimated that about 5–10 million people
ISSN 1658-6050
chew it every day (Feyissa and Kelly,
2008; Numan, 2012). In Yemen for example, 60% of the males and about 35% of
the females were found to be khat users
and had chewed daily for long periods of
their lives (Kennedy et al., 1980; Kennedy
et al., 1983). The chewing of khat leaves
has a stimulating effect and causes a certain degree of euphoria (Alsanusy and ElSetouhy, 2013; Gaffar et al., 2013; Kalix,
2008). Since only fresh leaves have the
desired effect, the chewing habit until the
present time has remained in those areas
where the plant is indigenous. The active
ingredient of Khat is cathinone (Patel,
2000; Wood et al., 2010). Cathinone is
unstable and undergoes decomposition reactions after harvesting and during drying
or extraction of the plant material. Decomposition leads to a dimer (3,6dimethyl-2,5-diphenylpyrazine) and posE-mail: siddigroa@yahoo.com
Journal of Jazan University - Applied Sciences Branch
Vol.3 No.1 Jan. 2014 (Safar 1435 H) 41
M.M. Halawi et al.,
sibly to smaller fragments. Both the dimer
and phenylpropanedione have been isolated from khat extracts (Wabe, 2012). As
cathinone is presumably the main psychoactive component of khat, this explains
why fresh leaves are preferred by the users.The phenylalkylamine content of khat
leaves varies within quite wide limits. On
average, 100 g fresh khat leaves contain
36–114 mg cathinone, 83–120 mg cathine
and 8–47 mg norephedrine (Balint et al.,
2009; El-Sayed and Amin, 2012; Vekariya, 2012). The effects on the nervous system resemble those of amphetamine, with
differences being quantitative rather than
qualitative (Brenneisen et al., 1990; Kalix,
1980). A previous study has found that the
main (psychoactive) compounds in khat
leaves are cathine and cathinone, which
are some to 2- to 10-fold less active than
amphetamine (Balint et al., 2009; Balint
and Balint, 1994).
Khat use affects also almost the whole
human organism. The main toxic effects
include increased blood pressure, tachycardia, insomnia, anorexia, constipation,
general malaise, irritability, migraine and
impaired sexual potency in men (AlBadri
et al., 2012; Alsalahi et al., 2012; Gezon,
2012; Nyachieo et al., 2012). It has been
reported that about 50% of khat chewers
develop oral mucosal keratosis. This pathologic change is considered a precancerous lesion that may develop into
oral cancer. The prevalence of this lesion
and its severity increased with frequency
and duration of khat use. In a survey in
Saudi Arabia about 50% of the patients
with head and neck cancer presented with
a history of khat chewing and all of them
had used khat over a period of 25 years or
more . In some cases the malignant lesion
occurred at exactly the same site as where
the khat bolus was held. The authors concluded that a strong correlation existed between khat chewing and oral cancer (Hal-
boub et al., 2012; Helal et al., 2012; Kassim and Croucher, 2012).
Serum proteins are proteins found in
the blood plasma, the clear, protein-rich
fluid which is left behind when platelets,
red blood cells, and white blood cells are
removed from the blood. These proteins
play a number of important roles in the
human body, and levels of Serum proteins
are sometimes evaluated in a laboratory
analysis to gather information about a patient's general health and specific health
issues which a patient may be experiencing (Anderson, 2010). There was no previous studies on the effect of khat on serum protein electrophoresis. Due to the
habitual widespread use of khat in Jazan
area, the present study was undertaken to
evaluate the effects of khat on the protein
electrophoresis pattern in khat chewing
males.
2. SUBJECTS AND METHODS
2.1.Subjects
After obtaining ethical approval from
the Medical Research Centre, Jazan
Univeristy Committee, recruits were enrolled. The ages were ranging between 2050 years and all were males. Venous
blood samples were drawn from the antecubital vein at the end of the khat chewing
session. A total of 70 blood samples were
collected from different localities from
Jazan. Fifty blood samples were taken
from subjects chewing khat and 20 samples from those who had no experience of
khat chewing and serve as a control group.
Samples were centrifuged (3500g, 10
minutes), and serum was separated and
freshly analyzed.
2.2. Serum Protein Electrophoresis
(Helena):
Helena SPIFE 3000 (serum proteinimmunofixation electrophoresis system
3000) Spilt Beta SPE System employedagarose gel in a tris-barbital MOPS buffer
42 Journal of Jazan University - Applied Sciences Branch
Vol.3 No.1 Jan. 2014 (Safar 1435 H)
Serum Protein Electrophoretic Pattern Among Khat Chewers
with calcium lactate for the separation of
serum protein upon electrophoresis into
six fractions. The semi-automated SPIFE
3000 system employed for electrophoresis
also performed the tasks of staining, destaining, and drying of the gels. The six
fractions after SPE were quantified us-
ingthe Helena scanner (QuickScan 2000).
Typical normal pattern for serum protein
electrophoresis is shown in Fig 1. Helena
SPIFE 3000 was gratefully provided by
King Fahd Central Hospital, Jazan, Saudi
Arabia.
Fig.1. Typical normal pattern for serum protein electrophoresis. Albumin, the largest
peak, lies closest to the positive electrode. The next five components (globulins) are labeled
alpha1, alpha2, beta1, beta2, and gamma. The peaks for these components lie toward the
negative electrode, with the gamma peak being closest to that electrode. Fig. 1 shows a typical normal pattern for the distribution of proteins as determined by serum protein electrophoresis.
2.3.Statistical analysis
Comparative study of the previous results of both groups were analysed using
the student t test and p value < 0.05 was
taken as significant.
3. RESULTS
The age of the subjects enrolled in
both groups (non-khat and khat users) was
ranging between 20-50 years and the subjects were males. The results are sum-
marized in Table 1 and Fig.2,3 & 4. Among the 50 khat chewing peoples 15 cases
(30%) showed serum albumin level above
the normal range whereas total protein
level was found to be elevated in
(16.66%) of cases. The levels of α-2 and
β-globulin in sera were found to be in
normal range. For, α-1 globulin, it is increased in 31.53% of khat users,while γ globulin showed 35.53 % decrease in khat
subjects.
Journal of Jazan University - Applied Sciences Branch
Vol.3 No.1 Jan. 2014 (Safar 1435 H) 43
M.M. Halawi et al.,
Table 1: Serum level and percent changes of serum proteins in both groups
1.
2.
3.
4.
5.
6.
Parameter
Non-khat users
khat users
% change
Total Protein
Albumin
α1-Globulin
α2- Globulin
β- Globulin
γ- globulin
7.72 ± 0.15
3.341 ± 0.39
2.199 ± 0.144
7.884 ± 0.256
8.748 ± 0.284
11.842 ± 0.644
8.86 ± 0.316
4.705 ± 0.41
2.365 ± 0.139
8.096 ± 0.298
8.917 ± 0.307
10.028 ± 0.596
16.66%
33.71%
31.53%
7.69%
5.12%
-35.53%
Results are expressed as Mean ± S.D. P< 0.05 was taken as significant.
Fig. 2. Perecentage of change in control and khat users in total protein an albumin
_
Fig. 3. Perecentage of change in control and khat users in globulin alpha1 and alpha2
44 Journal of Jazan University - Applied Sciences Branch
Vol.3 No.1 Jan. 2014 (Safar 1435 H)
Serum Protein Electrophoretic Pattern Among Khat Chewers
Fig. 4. Perecentage of change in control and khat users in beta and gamma globulin
4.DISCUSSION
Serum Protein electrophoresis involves the systematic separation, quantitation
and identification of the protein present in
the biological samples. By comparing the
proteins present in a diseased samples
with those in control samples, it is possible to identify the differences in the protein expression. Those differences may be
related to the response to organ toxicity or
to risk factors pre-existing before the acquisition of the disease (Cray et al., 2007;
Matt et al., 2008). With this aim, we have
carried out a prospective study on the differences of protein expression in sera between a control and khat chewing subjects. Our study suggested that serum albumin and α-1 globulin showed enhancement in subjects chewing khat than the
non-chewers, while γ-globulin was decreased due to khat chewing.
With the increasing evidence of the
deleterious effects of khat on human health and the social problems associated
with its use (Al-Habori, 2005), the levels
of serum proteins of khat users and nonusers were considered to find out the associated risk factor. Chewing of khat
leaves releases cathinone, a stimulant that
produces the feeling of euphoria. The release of cathinone in plasma showed that
Khat chewing induce significant increase
in blood pressure (Alkadi et al., 2002).
Long term use of khat or abuse can cause
insomnia, anorexia, gastric disorders, depression, liver damage and cardiac complications, including myocardial infarction
(Balint et al., 2009). On the basis of the
scientific data it seems clear that khat use
has negative consequences on the economic development of a country and on
the health of the society (Elmi et al., 1987;
Hansen, 2013). As far as we know, this is
the first paper that estimates the protein
fractions by electrophoresis. That is why
there is no previous data dealing with protein electrophoresis in khat chewing people.
Nincini et al., 1982 and 1984, discovered that consumption of Catha edulis and
cathinone increases plasma adrenocorticotrophic hormone (ACTH) in humans
(Ashenef and Engidawork, 2013; ElSayed and Amin, 2012). The stimulatory
effect of ACTH is believed to be mediated
by the activiation of adenyl cyclase through a membrane-bound receptor, with a
subsequent increase in cyclic AMP concentrations. The latter may then stimulate
various enzyme systems involved in enhancing the levels of serum proteins.
Moreover, Hyperalbuminemia is a relative
Journal of Jazan University - Applied Sciences Branch
Vol.3 No.1 Jan. 2014 (Safar 1435 H) 45
M.M. Halawi et al.,
change seen with sever dehydration condition and/OR poor protein utilization. It is
possible that a retinol (vitamin A) deficiency alone could cause albumin levels to
become raised. Hyperalbuminemia is also
associated with high protein diets. Increased α-1 globulin may be a sign of
acute or chronic inflammation (Bhonsle et
al., 2012). Gammaglobulins are decreased
in a variety of genetic immune disordes
and in secondary immune deficiency related to steroid use, leukemia or severe infection (Dasanu, 2012).
5.CONCLUSION
This study demonstrates that chewing
of khat, induced some differences in serum albumin, α-1 and γ- globulins. Extension of this study is needed on a large
scale to get more information about the effect of khat chewing on the protein status
and immune system in khat users.
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‫‪48 Journal of Jazan University - Applied Sciences Branch‬‬
‫)‪Vol.3 No.1 Jan. 2014 (Safar 1435 H‬‬
‫‪Serum Protein Electrophoretic Pattern Among Khat Chewers‬‬
‫نمط الترحيل الكهربائي لبروتينات دم مخزني القات‬
‫محمود حسين حلوي‪ ،1‬هشام إبراهيم أبو طويل‪،1‬‬
‫‪2‬‬
‫رشاد بن محمد السنوسي‪ ،2‬صديق إبراهيم عبد الوهاب‬
‫‪1‬‬
‫مركز األبحاث الطبية ‪ -‬جامعة جازان ‪ -‬المملكة العربية السعودية‬
‫مركز أبحاث المؤثرات العقلية ‪ -‬جامعة جازان ‪ -‬المملكة العربية السعودية‬
‫‪2‬‬
‫‪Q‬‬
‫تعرف أوراق القات بأن لها آثار محفزة وتعطي اإلحساس بالمتعة لمستخدميها‬
‫وتمضغ عادة من قبل الناس في شرق أفريقيا وبعض شبه الجزيرة العربية‬
‫واليمن وجنوب المملكة العربية السعودية‪ .‬نظرا ً النتشار استخدام القات في‬
‫منطقة جازان فقد أجريت هذه الدراسة لتقييم تأثير القات على نمط الكهربائي‬
‫للبروتين (ألفا) المعزول من دم الذكور الذين عندهم عادة مضغ القات‪ .‬تمت‬
‫هذه الدراسة علي مجموعة مكونة من ‪ 70‬شخص و التي كان مستخدمو القات‬
‫‪ 50‬و‪ 20‬من غير المستخدمين‪ .‬تم تحليل إجمالي بروتين الدم والزالل‬
‫والجلبويولين ألفا (‪ 1‬و‪ )2‬وبيتا وقاما‪ .‬وقد أظهرت النتائج أن نسبة إجمالي‬
‫البروتين قد ارتفعت بنسبة ‪ %16.66‬والزالل بنسبة ‪ %33.7‬والجلبويولين‬
‫بنسبة ‪ %31.5‬من بين إجمالي العينات‪ .‬فيما انخفضت نسبة القاما جلبويولين‬
‫بنسبة ‪ .%35‬اظهرت هذه الدراسة والتي تعتبر االولي من مثيالتها أن نمط‬
‫الترحيل الكهربائي لبروتينات الدم ونسبها قد تأثرت كأحد اآلثار الجانبية‬
‫السالبة المصاحبة لمضغ القات‪.‬‬
‫الكلمات المفتاحية‪ :‬مضغ القات‪ ،‬آلية الترحيل الكهربائي‪ ،‬نمط بروتينات الدم‬
‫باستخدام الترحيل الكهربائي‪.‬‬
‫‪E-mail: siddigroa@yahoo.com‬‬
‫‪ISSN 1658-6050‬‬