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Ameliorating Potentials of Lemon Zest on Pons Neurons in Canabis sativa-

Induced Wistar Rat

Article  in  Asian Journal of Advanced Research and Reports · April 2023

DOI: 10.9734/ajarr/2023/v17i6485

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 Asian Journal of Advanced Research and Reports

Volume 17, Issue 6, Page 17-23, 2023; Article no.AJARR.98386

ISSN: 2582-3248

Ameliorating Potentials of Lemon Zest

on Pons Neurons in Canabis sativa-
Induced Wistar Rat
Leyira Kenneth Yorkum a
and Ibinabo Fubara Bob-Manuel a*
a
Department of Anatomy, Faculty of Basic Medical Sciences, University of Port Harcourt, Choba,
Port Harcourt, Rivers State, Nigeria.

Authors’ contributions

This work was carried out in collaboration between both authors. Both authors read and approved the
final manuscript.

Article Information
DOI: 10.9734/AJARR/2023/v17i6485

Open Peer Review History:

This journal follows the Advanced Open Peer Review policy. Identity of the Reviewers, Editor(s) and additional Reviewers,
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Received: 15/02/2023
Accepted: 17/04/2023
Original Research Article
Published: 22/04/2023

ABSTRACT

Cannabis sativa, commonly known as marijuana has been indicated for many organs malfunction. It
is a common plant ingested or taken by people or individuals for body enhancement. The purpose
of this study is to determine the effects of lemon zest on the pons of cannabis sativa induced Wistar
rats. The plant was collected from National Drug Law Enforcement Agency (NDLEA), Rivers State,
Nigeria and 40 healthy Wistar rats were used for the experiment. The rats were divided into five (5)
groups. Group 1 served as control, groups 2,3,4 and 5 served as 250 mg/kg lemon zest only,
750mg/kg cannabis sativa only, 500 mg/kg cannabis sativa + 250 mg/kg lemon zest and 900mg/kg
cannabis sativa +250 mg/kg lemon zest respectively. The plant extract was administered for 21
days, and thereafter, the organs were harvested for routine histological tissue processing. Findings
-1
from the study revealed that when 500mgkg of Cannabis sativa was used to induce the pons,
observation showed neuronal hypertrophy and degeneration with mobilization of several supporting
cells. The connective tissue indicated an edematous appearance. It was also observed that when a
_____________________________________________________________________________________________________

*Corresponding author: Email: [email protected];

Asian J. Adv. Res. Rep., vol. 17, no. 6, pp. 17-23, 2023
 Yorkum and Bob-Manuel; Asian J. Adv. Res. Rep., vol. 17, no. 6, pp. 17-23, 2023; Article no.AJARR.98386

low dosage (500 mgkg-1) of the cannabis sativa was administered alongside 250 mgkg-1 extract of
lemon zest, well delineated neuronal cells with well-defined nuclei and nucleolus were observed in
-1
the tissue. When the dosage of the Cannabis sativa was increased to 900 mgkg alongside
250mgkg-1 extract of lemon zest, pockets of edematous connective tissues and blood vessel
congestion were still noticeable in the tissue. There was a general recovery (proliferation) of lost
cells in groups 4 and 5 respectively, as a consequence of the administration of lemon zest with
extract (Cannabis sativa) on Wistar rat. The improved tissue and cellular appearance are
indications of the potency of lemon zest in the amelioration of adverse tissue effects introduced by
the extract of Cannabis sativa.

Keywords: Cannabis sativa; pon; lemon zest; Wistar Rats.

1. INTRODUCTION Lemon zest contains high amounts of nutrients

and provides a wider range of health benefits
In the health of individuals and communities, when consumed as juice [5]. Mainly, the
medicinal plants and fruits have several vital cultivation of lemon is done for the alkaloids
roles. The medicinal values of these plants and which possess anticancer activities and the
fruits are possible due to the numerous chemical antibacterial activities in crude lemon-parts (such
substances contained in them which include as flowers, roots, stems, and leaves) extract and
phenolic compounds, oleoresins, resins, alkaloids also exhibit clinically noteworthy in
sesquiterpene, flavonoids, saponins glycosides, bacterial-strains. The notable part of lemon peel
alkaloids, fats, and oils, which on the body is the lemon zest. Lemon zest is the outermost
produce physiological action [1]. Over a long layer of the lemon peel known as the flavedo and
period, naturally occurring constituents used contains loads of natural oils that are used for
which has the abundant potential for yielding new various purposes including an influence on the
drugs, and the constituents contain a biologically nervous system as well as influence on Cannabis
active substance that demonstrates the biological Sativa [6-9].
activity for human health. The naturally occurring
The study is aimed to determine the effects of
antimicrobial substance has greater significant
lemon zest on the pons of cannabis sativa
values and has more importance in plant
induced Wistar rats.
chemotherapy and these substances are found
in plants and fruits such as lemon [2]. 2. MATERIALS AND METHODS
Cannabis sativa, commonly termed marijuana, Experimental Design: This study will adopt the
weed, pot, and ganja, is one of the earliest Completely Randomized Design (CRD). This
known cultivated plants. Its cultivation started involves two principles which are the principles of
with the birth of agricultural farming around replication and the principle of randomization of
10,000 years ago [3]. It is a multi-purpose crop experimental design.
plant with diverse agricultural and industrial
applications which range from the production of Collection and Accumulation of Cannabis
paper, wood, and fibre to pharmaceutical uses in Sativa: Dried leaves, twigs and seeds of
industries. Tajammul, et al. [3] claimed that the cannabis sativa plant were obtained from the
first prospect of cannabis sativa as a medical National Drug Law Enforcement Agency
plant was published in 1843 where its use was (NDLEA), Rivers State command, Nigeria. The
described for treating patients suffering characterization and validation of the plant will be
from tetanus, hydrophobia, and cholera. done by evidence specialists at NDLEA, Rivers
Consequently, the plant was removed from the State Command in Port Harcourt.
medicinal category and recategorized exclusively Animal Collection and Acclimatization: Forty
to the category of drug-type plants. (40) Wistar rats of both sexes were used in this
study. The rats were obtained from the animal
Niloy, et al. [4] focus on the effects of cannabis house in the Faculty of Basic Medical Sciences,
on the different cognitive domains, including University of Port Harcourt, and kept in clean
learning, consolidation, and retrieval. Their study cages under standard laboratory conditions of 12
is the first attempt in which significant focus has hours light/dark cycle. They had access to vital
been imparted on all three aspects of cognition, pellet feed and distilled water as they were
thus linking to its usage. allowed to acclimatize for one week. The rats

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 Yorkum and Bob-Manuel; Asian J. Adv. Res. Rep., vol. 17, no. 6, pp. 17-23, 2023; Article no.AJARR.98386

were allowed to roam in an enclosed space Tissue Harvesting: The harvesting of tissues
having proper ventilation and temperature was done to extract the organ of choice from the
throughout the experiment. All the rats were rat which is the pons. The rats were first made
allowed to feed ad libitium for 14 days after which unconscious by being exposed to chloroform.
different behavioural tests will be conducted. While unconscious, the rats were then pinned to
a wooden board to keep them still during
Experimental Procedure: The 40 Wistar rats dissection. The brains were harvested from their
with an average weight of 110g were randomly skull and the harvested parts were fixed in Bonin
divided into 5 groups (8 rats each). A low dose solution.
and high dose of 250mg/kg and 900mg/kg
respectively and a normal dose of 750mg/kg of 3. RESULTS
aqueous marijuana were used according to
research conducted by Danladi, et al. [10] to The Plate 1 below shows the photomicrograph of
induce the rat. The dosage of lemon zest used the pons used for the control experiment at lower
-1
includes 250mgkg following the findings of magnification showing large pyramidal neuronal
Juan, et al. [11]. cell bodies at the anteromedial area of the
pons. Medium size neuronal cell bodies are
Group 1: Control group- This group was given
arranged at the posterior medial area of the
animal feed and distilled water only with no dose
pons. Tissue show normal cytoarchitecture. H &
of lemon zest or marijuana.
E, X100.
Group 2: Low dose group- This group was
served with a low dose (250 g) of lemon zest Plate 2 below displays the photomicrograph of
only and no dose of marijuana. the pons showing well delineated neuronal
cell bodies and few supporting cells (glial). The
Group 3: Marijuana Induced group: This group connective tissue fibres are well distributed. H &
was administered a 500 mg/kg dose of Marijuana E, X400.
and no dose of lemon zest.
Group 4: This group was induced with The Plate 3 below shows the photomicrograph
-1 of the pons on few degenerating neuronal cell
500mgkg of marijuana and treated with low
-1 bodies and numerous supporting cells (glial)
dose (250 mgkg ) of lemon zest. -1
after 250mgkg of lemon zest was administered.
Group 5: This group was served with a high Pockets of edematous connective tissue cells
dose (900g) of marijuana but with low dose (250 are observed. Pons tissue show mild distortion.
-1
mgkg ) of lemon zest Lemon. H & E, X400.

Plate 1. Normal control showing large pyramidal neuronal cell bodies

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 Yorkum and Bob-Manuel; Asian J. Adv. Res. Rep., vol. 17, no. 6, pp. 17-23, 2023; Article no.AJARR.98386

Plate 2. Normal control showing delineated neuronal and supporting cells

-1
Plate 3. Pons of the Wistar rats after 250mgkg of lemon zest induction

Plate 4 below is the photomicrograph of the pons observed. Pons shows a recovery state of tissue
showing several hypertrophied and degenerating cytoarchitecture. H & E, X400.
neuronal cell bodies alongside numerous
supporting cells (glial-arrowed) when a low dose 3.1 Effects of Lemon Zest on the
-1
of 250mgkg of cannabis sativa was Histology of Pons of High Dose
administered to the rat. Diffused edematous Cannabis Sativa Induced Wistar Rat
connective tissue cells are observed. Pons
shows severe distortion of tissue Plate 6 below displays the photomicrograph of
cytoarchitecture. H & E, X400. the pons showing proliferation and degenerating
pyramidal neuronal cell bodies with pockets of
The Plate 5 below displays the photomicrograph supporting cells (glial). Some well delineated
of the pons showing proliferation of pyramidal neuronal cell bodies with clear nucleolus are
neuronal cell bodies with pockets of supporting outlined in the tissue. The presence of congested
cells (glial). Pockets of edematous (vacuolation) blood vessels containing lymphocytic cells is also
connective tissue cells (arrowed) and the observed. Pons shows a recovery state of tissue
presence of congested blood vessel are also cytoarchitecture. H & E, X400.

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 Yorkum and Bob-Manuel; Asian J. Adv. Res. Rep., vol. 17, no. 6, pp. 17-23, 2023; Article no.AJARR.98386

-1
Plate 4. Pon state after the medium dose of Cannabis Sativa (500 mgkg ) induction only

-1 1
Plate 5. Low dose of Cannabis Sativa (250 mgkg ) and a low dose of lemon zest (250 mgkg )
application

-1 1
Plate 6. High dose of Cannabis Sativa (900 mgkg ) and a low dose of lemon zest (250 mgkg )
application

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 Yorkum and Bob-Manuel; Asian J. Adv. Res. Rep., vol. 17, no. 6, pp. 17-23, 2023; Article no.AJARR.98386

4. DISCUSSION lemon zest has neuroprotective effects in

individuals since the Wistar rats’ brain is similar
This study aims at determining the effects of to that of the humans. The neuroprotective
lemon zest on the pons of cannabis sativa effects observed in this present study agrees
induced Wistar rats. The study thus determined with the findings of Shafeeqa et al. [17].
the effects of lemon zest on the cytoarchitecture
of pons of cannabis sativa non-induced Wistar 5. CONCLUSION
rats, determined the effects of a low dose of
lemon zest on the histology of pons of cannabis Conclusively, this study has demonstrated the
sativa induced Wistar rats and examined the negative effect of cannabis sativa on the
impacts of high dosage of lemon zest on neuronal cells of the pons upon administration of
histology pons of cannabis sativa induced Wistar different doses of the extract, as demonstrated in
rats. group 3. However, this effect was reversed upon
administration of lemon zest as established in
Results in this study displayed the groups 4 and 5. It is therefore important to note
photomicrograph of the pons before marijuana that lemons zest has neuroprotective qualities or
and/or lemon zest were administered. Findings the potential to ameliorate adverse effects
-1
thus revealed that when 250 mgkg of lemon caused by the extract of Cannabis sativa.
zest was administered, pons showed few
degenerating neuronal cell bodies as well as
COMPETING INTERESTS
numerous supporting celles (glaila) as against
the normal control which displayed well
delineated neuronal cell bodies and few Authors have declared that no competing
supporting cells since the connecting tissues interests exist.
-1
were properly distributed. When 500 mgkg of
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