Isoniazid is an antibacterial that works by
inhibiting bacterial growth. It is most commonly used for the treatment of
tuberculosis. The inflammation process is a complex defence
mechanism of the body which occurs as a response to trauma, heat, chemicals, bacteria
and/or virus. Annonamuricata is
a medicinal plant widely used in traditional medicine for its antibacterial,
antioxidant and anti-inflammatory properties. This study investigated the
effects of Annonamuricata, a
medicinal plant, on inflammation and toxicity following Isoniazid
administration. Forty four male wistar rats were
used in the study. Two hundred and fifty grams of dried Annonamuricata leaf was macerated in 1000 mls of 95% absolute ethanol for 48-hours and was
administered orally at a dose of 100 mg/kg, 500 mg/kg and 1000 mg/kg.
Inflammation was induced by injecting 0.5 ml of egg white in the hind paws
and was observed at an interval of thirty minutes. 150 mg/kg of Isoniazid was
administered intraperitoneally thirty minutes after
extract administration and then observed for two hours for convulsion and
death. Data was analysed using ANOVA followed by
post hoc Fisher’s LSD and values were considered significant at p<0.05.
SEM. The findings of the study revealed a significant decrease in AST levels,
liver weight, seizure and itching levels. It also demonstrated a significant
decrease in paw sizes at 60-120 minutes. However, further experiments should
be carried out to confirm the findings of this study and the mechanism behind
the decrease in seizure and itching levels.
Accepted:20/06/2024
Published: 03/07/2024
*Corresponding Author
Dr Emmanuel NonsoEzeokafor
E-mail: e.ezeokafor@ unizik.edu.ng
Tel: +234 803 098 7212
Keywords: Isoniazid, Annonamuricata, Wistar Rats
INTRODUCTION
Inflammation is a complex process. It occurs as a
response to trauma, heat, chemicals, bacteria or other phenomena and is
mediated by a variety of electrically charged signal molecules produced locally
by mast cells, nerve endings and platelets (Hardset al.,1994). Characteristics include;
heat, redness, swelling, pain and loss of function. Inflammation is a part of
the defence mechanism of the body however, it has the
potential to become harmful and uncontrolled at times like in rheumatoid
arthritis and many other autoimmune inflammatory conditions (Chen et al., 2017).
A convulsion occurs when nerve cell activity in the
brain is disrupted, causing muscles to involuntarily contract and spasm,
resulting in sudden, violent, and irregular movements of the body (Moawad, 2022). It may be associated with any number of
medical conditions, including epilepsy, a head injury, severe fever, an
inflammatory brain infection, exposure to toxins, and certain medications.
Diagnosis by a doctor such as a neurologist or infectious disease specialist is
required to determine the reason it has occurred and to ascertain the
appropriate treatment plan (Stafstrom and Carmant, 2015).
Isoniazid is a drug of choice for the treatment of
pulmonary and extrapulmonary tuberculosis, and is
used for the chemoprophylaxis of tuberculosis. Typically it is used in
combination with other antituberculosis agents to
prevent or delay the appearance of resistance. It is one of the few drugs that
is bactericidal for all populations of the tuberculosis organism. Isoniazid is
a prodrug. It acts by inhibiting the formation of the
mycobacterial cell wall. Isoniazid must be activated by KatG,
a bacterial catalase-peroxidase enzyme in Mycobacterium tuberculosis (Suarez et al., 2009).
A plant is said to be medicinal when its biologically
active compounds have shown considerable pharmacological activities such as
antioxidant, antimicrobial, anti inflammatory,
anticancer, antiviral, anti-allergic and vasodilatory
properties (Rustaiyanet al., 2011). An example is AnnonaMuricata. The plant derived herbal compounds have a long
history of clinical use, better patient tolerance and acceptance. Their high
ligand binding affinity to the target introduces the prospect of their use in
chemo preventive applications; in addition they are freely available natural
compounds that can be safely used to prevent various ailments. Plants became
the basis of the traditional medicine system throughout the world for thousands
of years and continue to provide mankind with new remedies.
MATERIALS
AND METHOD
Location of
the study: This study was carried out
in the animal house, department of Human Physiology, Faculty of Basic Medical
Sciences, College of Health Sciences, Nnewi campus, NnamdiAzikiwe University.
Materials: 44 male wistar rats,
laboratory coat and gloves, rice dusts, centrifuge (search tech instruments, british standard) model 80-2, thermostat oven dhg-90 23a, pec medical, USA, rotary evaporator (digital) tt-52 techmel and techmelusa, thermostatic water bath, beakers and measuring
cylinders, 2ml hypodermic syringes, plain bottles, electronic weighing balance,
m-methlar model m311l, China, watt-man number1 filter
paper, refrigerator (haierthermocool),
cages, oral cannula, rat feeds (vital pelleted finisher), egg white, AISY
Absorbent Cotton Wool(Hydrophilic), 95% Absolute Ethanol AR JHD: (GuangdangGuanghua Chemical factory,China), Chloroform AR JHD: (GuangdangGuanghua Chemical factory, China).
Plant Sample
Collection and Identification: The
leaves of soursop were harvested from a local farm in
Okofia, Nnewi LGA of Anambra state. It was sent to the Botany Department Awka where it was identified by a botanist and the soursop leaves were registered as NAUH-04B. Shade dried
leaves were pulverised into fine powder using an
electric blender and stored in an air-tight container for further use.
Preparation
of Ethanolic Extract of AnnonaMuricata: The
Annonamuricata
leaves were harvested from a local farm in Okofia, Nnewi Local Government Area, Anambra State. The Annonamuricata leaves were dried at an ambient
temperature. The dried Annonamuricata
leaves were milled into a coarse form using a local grinder. Two hundred and
fifty-(250) grams of the Annonamuricata leaf
was macerated in 1000mls of 95% absolute ethanol (JHD Chemicals, Guangdong, China) for 48-hours. It was then filtered using a clean
porcelain cloth and further filtered using Filter paper (Whatman
Qualitative Filter paper, No. 1, Sigma Aldrich; WHA1001042, USA). The filtrate
obtained was concentrated using a rotary evaporator (Digital TT-52; Techmel&Techmel, USA), which was further dried using a
laboratory oven (DGH-9023A, PEC MEDICAL, USA) at 45şC into a gel-like form. The
extract was preserved in a refrigerator (Haier Thermocool)
for further usage. The extract procedure was done according to the method described
by Attar and Abu-Zeid (2013).
Preparation of Isoniazid Solution and
Induction of Convulsion
The effect of ethanolicAnnonamuricata
leaves extract on Isoniazid -induced convulsion was assessed as described by (Cordaet al.,
1982). The animals were divided into four groups and were given (100, 500 and
1000 mg/kg) of ethanolicAnnonamuricata leaves extracted orally
respectively. Thirty minutes later, the rats in all groups were given Isoniazid
(150 mg/kg) intra peritoneally and were observed for
two hours for convulsion and death.
Induction of Inflammation Using Egg
White
Inflammation was induced by injection of 0.2ml of egg
white in hind paw 30 minutes before administration of ethanolic
extract of Annonamuricata
leaves; the size of individual hind paw was determined using verniercalliper, which was
observed at 30 minutes interval for a period of 2 hours. The hind paw of the
rats in group A, B, C, D and E were measured using a verniercalliper and recorded before the egg white was
injected into the hind paw. This was done according to the method described by
(Barunget al.,
2021).
Experimental
Animals
Experimental rats were purchased from the animal house
in the department of Veterinary Medicine in the University of Nigeria (UNN) Nsukka. The animals weighed between 160-300g, and were
randomly grouped and housed in steel cages and kept at room temperature. The
rats had no history of drug consumption, that is; they had not been used for
any investigation. The rats were put on standard rat's pellet (feed) and pure
drinking water and allowed to get acclimated for 21 days before the start of
the experiment. The study was done in accordance with the guidelines for animal
use of the Faculty of Basic Medical Sciences, NnamdiAzikiwe University.
Experimental
Design
Forty four male wistar rats
were divided into two different research subgroups; anti-inflammatory subgroup
and anti-convulsant subgroup. To check for
anti-inflammatory properties of Annonamuricata, twenty five male wistar
rats were used while to check for the anticonvulsant properties of Annonamuricata,
twenty male wistar rats were used.
Anti-inflammatory
effect of ethanolic extract of annonamuricata leaves.
Twenty four male wistar rats
were divided into six groups of four, the anti-inflammatory subgroups were
designated as groups A, B, C, D, E and F.
·Group A
(negative control): (0.2ml) egg white
·Group B (Positive control): Feed
+ distilled water (10 ml/kg).
·Group C (Low dose): (0.2ml) egg
white + extract 100ml.
·Group D (Medium dose):(0.2ml) egg white + extract 200ml.
·Group E (High dose):(0.2ml) egg white + extract 400ml.
·Group F (Very high dose):(0.2ml) egg white+ extract 800 ml.
The
hind paw of the rats in group B, C, D and E were measured using a verniercalliper and recorded
before the egg white was injected.
Anticonvulsant effect of ethanolic extract of annonamuricata leaves.
Twenty
male wistar rats were divided into four groups of
five, the anticonvulsant subgroup were designated as groups A, B, C, and D.
·Group A
(negative control): (150mg) Isoniazid
·Group B (low dose): (150mg)
Isoniazid + (extract 100ml for seven days).
·Group C (medium dose): (150mg)
Isoniazid + (extract 500ml for seven days).
·Group D (high dose): (150mg)
Isoniazid + (extract 1000ml for seven days).
RESULTS
Table 1: effect of ethanolic
leaf extract of Annonamuricata on
liver enzymes (AST, ALT, and ALP) following Isoniazid induced hepatotoxicity
Aspartate
Transaminase (IU/L)
Alanine Transaminase
(IU/L)
Alkaline Phosphatase
(IU/L)
MEAN±SEM
MEAN±SEM
MEAN±SEM
Group A (150 mg/kg of Isoniazid)
49.00±1.00
36.33±4.33
78.53±1.88
Group B (150 mg/kg of ISA + 100 mg/kg of
EAML)
37.67±0.67*
29.67±1.33 a
114.33±10.88 *
Group C (150 mg/kg of ISA + 500 mg/kg of
EAML)
38.33±2.96 *
35.33±0.88 a
114.23±13.77*
Group D (150 mg/kg of ISA + 1000 mg/kg of
EAML)
37.33±1.67*
38.00±5.29a
84.76±5.55 a
F-value
9.74
1.06
4.22
Data was analysed using
ANOVA followed by post hoc Fisher’s LSD and values were considered significant
at p<0.05. SEM: Standard error of
mean, significant (*) and
not significant (a),
EAML: ethanolic leaf extract of Annonamuricata. ISA: Isoniazid
Table 1 results showed a significant decrease in
groups B, C, and D (p=0.002, p=0.003,
p=0.002) in AST level when compared to group A. The ALT level showed a
non-significant decrease in groups B and C (p=0.216,
p=0.845) and group D had a non-significant increase (p=0.746) compared to group A. The ALP result showed a significant
increase in groups B and C (p=0.026,
p=0.026) while group D had a non-significant increase (p=0.647) compared to group A.
Table 2
effect of ethanolic leaf extract of Annonamuricata on
seizure, itching, and relative brain weight following Isoniazid induced
neurotoxicity
Seizure (Frequency)
Itching (Frequency)
Relative brain weight
(g)
MEAN±SEM
MEAN±SEM
MEAN±SEM
Group A (150 mg/kg of Isoniazid)
45.00±2.88
95.00±2.88
0.70±0.01
Group B (150 mg/kg of ISA + 100 mg/kg of
EAML)
26.67±1.67*
78.00±1.53 *
0.82±0.01 a
Group C (150 mg/kg of ISA + 500 mg/kg of
EAML)
14.00±0.57*
47.33±1.45 *
0.79±0.01 a
Group D (150 mg/kg of ISA + 1000 mg/kg of
EAML)
7.00±1.15*
25.00±2.88*
0.76±0.03a
F-value
87.06
184.88
6.59
Data was analysed using
ANOVA followed by post hoc Fisher’s LSD and values were considered significant
at p<0.05. SEM: Standard error of
mean, significant (*) and
not significant (a),
EAML: ethanolic leaf extract of Annonamuricata. ISA: Isoniazid
Table 2 results revealed a significant decrease in
groups B, C, and D (p=0.001, p=0.010,
p=0.000) in the seizure level when compared to groups A. The result of
itching showed a significant decrease in groups B, C, and D (p=0.001, p=0.002, p=0.001) compared to
group A. The relative brain weight showed a significant increase in groups B
and C (p=0.003, p=0.014), and group D
had a non-significant increase (p=0.059)
compared to group A.
Table 3
effect of ethanolic leaf extract of Annonamuricata on
relative liver weight following Isoniazid toxicity
Relative liver weight
(g)
MEAN±SEM
Group A (150 mg/kg of Isoniazid)
3.98±0.11
Group B (150 mg/kg of ISA + 100 mg/kg of
EAML)
4.19±0.09a
Group C (150 mg/kg of ISA + 500 mg/kg of
EAML)
3.76±0.01a
Group D (150 mg/kg of ISA + 1000 mg/kg of
EAML)
3.43±0.15*
F-value
10.04
Data was analysed using
ANOVA followed by post hoc Fisher’s LSD and values were considered significant
at p<0.05. SEM: Standard error of
mean, significant (*) and
not significant (a),
EAML: ethanolic leaf extract of Annonamuricata. ISA: Isoniazid
Table 3 results showed the relative liver weight had a
non-significant increase in group B (p=0.173),
group C had a non-significant decrease (p=0.173)
and group D indicated a significant decrease (p=0.001) compared to group A.
Table 4
effect of ethanolic leaf extract of Annonamuricata on
paw-size at 0 and 30 minutes following egg white induced inflammation
0-minutes (cm)
30-minutes (cm)
MEAN±SEM
MEAN±SEM
Group A (0.2 ml of
Egg white)
2.40±0.17
4.00±0.05
Group B (positive
control)
0.98±0.01*
1.08±0.01*
Group C (0.2ml of Egg
white + 100 mg/kg of EAML)
2.53±0.06*
3.93±0.12a
Group D (0.2ml of Egg
white + 200 mg/kg of EAML)
2.30±0.05 *
3.83±0.08a
Group E (0.2ml of Egg
white + 400 mg/kg of EAML)
2.53±0.08*
3.93±0.12 a
Group F (0.2ml of Egg
white + 800 mg/kg of EAML)
2.43±0.07*
3.76±0.37a
F-value
0.96
0.26
Data was analysed using
ANOVA followed by post hoc Fisher’s LSD and values were considered significant
at p<0.05. SEM: Standard error of
mean, significant (*) and
not significant (a),
EAML: ethanolic leaf extract of Annonamuricata.
Table 4 revealed a significant increase in the paw
size in groups A, C, D, E, and F (p=0.010,
p=0.001, p=0.012, p=0.010, p=0.002) compared to group B at zero minutes. At
30 minutes, a non-significant decrease in the paw-size was revealed in groups
C, D, E, and F (p=0.802, p=0.534,
p=0.802, p=0.389) compared to group A, while group B had a significant
increase compared to group A (p=0.010).
Table 5 effect of ethanolic
leaf extract of Annonamuricata on
paw-size at 60, 90 and 120 minutes following egg white induced inflammation
60-minutes (cm)
90-minutes (cm)
120-minutes (cm)
MEAN±SEM
MEAN±SEM
MEAN±SEM
Group A (0.2 ml of
Egg white)
3.76±0.07
3.63±0.08
3.60±0.15
Group B (positive
control)
1.18±0.01*
1.18±0.01*
1.18±0.01*
Group C (0.2ml of Egg
white + 100 mg/kg of EAML)
3.00±0.05*
2.63±0.27*
2.70±0.25*
Group D (0.2ml of Egg
white + 200 mg/kg of EAML)
2.90±0.21*
2.50±0.15*
2.33±0.18*
Group E (0.2ml of Egg
white + 400 mg/kg of EAML)
2.73±0.17*
2.20±0.05*
2.16±0.6*
Group F (0.2ml of Egg
white + 800 mg/kg of EAML)
2.80±0.30*
2.60±0.30*
2.36±0.26*
F-value
5.08
7.38
8.52
Data was analysed using
ANOVA followed by post hoc Fisher’s LSD and values were considered significant
at p<0.05. SEM: Standard error of
mean, significant (*) and
not significant (a),
EAML: ethanolic leaf extract of Annonamuricata.
Table 5 results showed a significant decrease in the
paw size at 60-minutes in groups C, D, E, and F (p=0.015, p=0.008, p=0.003, p=0.004) compared to group A. However,
group A showed a significant increase in the paw size compared to group B (p=0.020). At 90-minutes, a significant
decrease in the paw size in groups B, C, D, and E (p=0.005, p=0.002, p=0.000, p=0.004) compared to group A. However,
group A showed a significant increase in the paw size compared to group B (p=0.004). At 120 minutes, a significant
decrease in the paw size in groups B, C, D, and E (p=0.009, p=0.001, p=0.000, p=0.001) compared to group A. However,
group A showed a significant increase in the paw size compared to group B (p=0.012).
DISCUSSION
Inflammation is a complex biological response of the
body to various pathological stimuli like pathogens and toxic compounds (Chen et al., 2017). Egg-white has been proven
to induce inflammation in rat paws by causing edema formation however the
references to explain the mechanism behind it remains inadequate. Researchers
suspect that the proteins; ovalbumin (54%), ovotransferrin
(12%), ovomucoid (11%), ovomucin
(3.5%), and lysozyme (3.5%; Abeyrathne, Lee and Ahn, 2013) contained in egg white are the major allergens
and serve as the trigger for an inflammatory response (Kim et al., 2015).
Annonamuricatais a
tropical plant widely characterised for its
biological activities (antibacterial, anti-inflammatory, antioxidant, etc). Analysis of egg-white induced inflammation between
0-30 minutes in all groups when compared to the control group showed a
significant increase in paw size. This supports the study by Barung, Dumanauw, Duri and Kalonio, 2021 which
states the potency of egg-white as an edema inductor. No significant decrease
was observed in the groups treated with A.
muricata at 0-30 minutes however, a significant
decrease was observed in all treated groups at 60-120 minutes. This decrease
suggests the efficacy of A. muricata as an anti-inflammatory agent. The effect
could be attributed to the flavonoids and terpenoids
present in the fruit (Okeke-Nwolisa, Enweani-Nwokelo, Unekwe and Egbuonu 2023).
A seizure occurs due to an uncontrolled, abnormal
electrical activity of the brain that may cause changes in the level of
consciousness, behaviour, memory, or feelings (Huff
and Murr, 2023). A.
muricata has been used traditionally as an
anticonvulsant and a sedative. Analysis of seizure and itching levels of groups
treated with A. muricata
after Isoniazid administration showed a significant decrease suggesting the
effectiveness of the extract as an anticonvulsant. This agrees with the study
by Souza et al., 2018 which indicates a possible interaction between the
extract and the GABAergic and monoaminergic
systems of the brain possibly as a result of the flavonoids and phenolic acids
present.
AST, ALT and ALP are enzymes found in the liver and
are markers for hepatocellular damage. AST serves as a more sensitive marker
for liver damage because of its abundance when compared to ALT while ALT serves
as a more specific marker due to its restricted tissue expression (Jorge,
2013). Analysis of liver enzyme levels of treated groups following Isoniazid
induced toxicity showed a significant decrease in AST levels of all groups
compared to the untreated group. This suggests the possibility of A. muricata as
an ameliorative agent for hepatocellular damage. A significant increase was
also observed in ALP levels of groups B and C when compared to group A however,
the reason for this increase is not understood.
CONCLUSION
The study demonstrated that ethanolic
extract of A. muricata significantly decreased paw
size following egg white induced inflammation at 60-120 minutes. The mechanism
of this decrease is linked to the flavonoids and terpenoids
contained in the extract. A decrease in AST levels, liver weight, seizure and
itching levels were also noticed suggesting the possibility of an
anticonvulsant activity. The decrease in AST levels suggests that A. muricata could be hepatoprotective
which can be attributed to the annonaceousacetogenins it contains. Further experiments should be
carried out to confirm the findings of this study and the mechanism behind the
decrease in seizure and itching levels.
Acknowledgement
The
authors wish thank God for his goodness and appreciate everyone who contributed
in one way or the other to ensure this project was a success.
Conflict of Interest
The authors declare no conflict of interests.
REFERENCES
Abeyrathne, E. D., Lee, H. Y., and Ahn,
D. U. (2013). Egg white proteins and their potential use in food processing or
as nutraceutical and pharmaceutical agents--a review.
Poultry science, 92(12), 3292–3299.
https://doi.org/10.3382/ps.2013-03391
Al-Attar, A.M. and
Abu Zeid, I.M., (2013). Effect of tea (Camellia sinensis) and olive (Oleaeuropaea L). leaves extracts on
male mice exposed to diazinon. BioMed research international.
Barung, E.N., Dumanauw, J.M., Duri, M.F. and Kalonio, D.E.,
(2021). Egg white-induced inflammation models: A study of edema profile and
histological change of rat's paw. Journal
of Advanced Pharmaceutical Technology & Research, 12(2), p.109.
Corda M. G., Costa E., Guidotti
A., (1982). Specific proconvulsant action of an imidazobenzodiazepine (Ro 15-1788) on isoniazid
convulsions. Neuropharmacology,
21(1), 91–4. DOI: 10.1016/0028-3908(82)90217-9
Chen, L., Deng, H.,
Cui, H., Fang, J., Zuo, Z., Deng, J., Li, Y., Wang,
X., and Zhao, L. (2017). Inflammatory responses and inflammation-associated
diseases in organs. Oncotarget, 9(6), 7204–7218.
https://doi.org/10.18632/oncotarget.23208
Hards, A., Sekido, N., Akahosi, T., Mukaida, N., and
Matsushima, K., (1994). Essential involvement of interleukin-8 (IL-8) in acute
inflammation. Journal of leukocyte
biology, 56(5), pp.559-564.
Huff J. S, Murr N. I. Seizure. [Updated 2023 Feb 7]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2024 Jan-. Available from:
https://www.ncbi.nlm.nih.gov/books/NBK430765/
Jorge Sepulveda
(2013). Chapter 9 - Challenges in Routine Clinical Chemistry Analysis: Proteins
and Enzymes. Editor(s): AmitavaDasgupta,
Jorge L. Sepulveda. Accurate Results in the Clinical Laboratory, Elsevier,
Pages 131-148, ISBN 9780124157835.
https://doi.org/10.1016/B978-0-12-415783-5.00009-8.
Kim, J. H., Song,
H., Kim, H. W., & Lee, W. Y. (2015). Effects of Egg White Consumption on
Immune Modulation in a Mouse Model of Trimellitic
Anhydride-induced Allergy. Korean journal
for food science of animal resources, 35(3), 398–405. https://doi.org/10.5851/kosfa.2015.35.3.398
Okeke-Nwolisa, B. C., Enweani-Nwokelo, I.
B., Unekwe, P. C., and Egbuonu,
I. (2023). The anti-diarrheal and anti-inflammatory effects of hydroethanol extracts of ripe Annonamuricata fruit pulp in Wistar
rats using curative method. International
journal of health sciences, 17(3), 26–32.
Rustaiyan, A., Javidnia, K., Farjam, M.H., Aboee-Mehrizi, F.
and Ezzatzadeh, E., (2011). Antimicrobial and
antioxidant activity of the Ephedra sarcocarpa
growing in Iran. Journal of Medicinal
Plants Research, 5(17), pp.4251-4255.
Souza, D. O., Dos
Santos Sales, V., de Souza Rodrigues, C. K., de Oliveira, L. R., Santiago Lemos, I. C., de AraújoDelmondes, G., Monteiro, Á. B.,
do Nascimento, E. P., SobreiraDantasNóbrega de Figuęiredo, F. R., Martins da Costa, J. G., Pinto da Cruz,
G. M., de Barros Viana, G. S., Barbosa, R., Alencar de Menezes, I. R., Bezerra Felipe, C. F., and Kerntopf,
M. R. (2018). Phytochemical Analysis and Central Effects of AnnonaMuricata Linnaeus: Possible Involvement of the Gabaergic and Monoaminergic
Systems. Iranian journal of
pharmaceutical research: IJPR, 17(4), 1306–1317.
Stafstrom C. E., Carmant L., (2015).
Seizures and epilepsy: an overview for neuroscientists. Cold Spring HarbPerspect
Med, 5(6):a022426. doi:10.1101/cshperspect.a022426
Suarez J., Ranguelova K., Jarzecki A. A, Manzerova J., Krymov V., and Zhao
X., (2009). An oxyferrousheme/protein-based
radical intermediate is catalytically competent in the catalase reaction of
Mycobacterium tuberculosis catalase-peroxidase (KatG).
The Journal of Biological Chemistry.
284 (11): 7017-7029. doi:10.1074/jbc.M808106200. PMC 2652337. PMID 19139099.
Cite this Article: Ezeokafor, EN; Uchefuna, RC; Ebuka, IE; Kasarachi, OD; Ebisintei, P; Enendu, AC; Aralu, OC; Okoye, KP; Okafor, CE (2024).
Effect of Ethanolic Extract of Annonamuricata Leaves on Inflammation,
Seizure and Liver Enzymes Following Isoniazid Toxicity in Male Wistar Rats. Greener
Journal of Medical Sciences, 14(2): 94-100.
