Download Effects of Leaf and Root Extracts of Newbouldia laevis on Hepatic

367
Journal of Pharmaceutical, Chemical and Biological
Sciences
ISSN: 2348 -7658
Impact Factor (GIF ): 0.615
Impact Factor (SJIF): 2.092
September -November 2015; 3(3): 367 -372
Original Research Article
Effects of Leaf and Root Extracts of Newbouldia laevis on Hepatic
and Renal Systems in Albino Rats
Agbafor K. N*, Ezeali C.
Biochemistry Department, Ebonyi State University, Abakaliki, Nigeria
*Corresponding Author: Agbafor K. N, Biochemistry Department, Ebonyi State University, Abakaliki,
Nigeria
Received:11September 2015
Revised:19 September 2015
Accepted:23 September 2015
ABSTRACT
Traditional medicine practitioners in Eastern Nigeria use various parts of Newbouldia laevis in
treatment and management of several disorders. This research investigated possible toxicity of leaf
and root extracts of the plant to liver and kidney in albino rats. Extractions were performed with
deionized water and ethylacetate to produce deionized water leaf (DWL), deionized water root (DWR),
ethylacetate leaf (EAL) and ethylacetate root (EAR) extracts. A total of 85 adult male albino rats, used
in the study, were placed in 16 test and one control groups of five rats in each group. The test groups
were given oral administration of 200, 400, 600 and 800mg/kg body weight of the extracts, the control
received normal saline for 21 consecutive days. The activities of alanine aminotransferase (ALT),
aspartate aminotransferase (AST), alkaline phosphatase (ALP) and gamma glutamyltransferase (GGT)
in the serum were used to assess hepatobilliary toxicity, while serum creatinine, urea and uric acid
were used as renal toxicity indicators. The activities of ALT, AST, ALP and GGT decreased insignificantly
(P>0.05) in the groups given 200 and 400mg/kg of DWL extract, while the enzymes activity groups
given 200 and 400mg/kg of other extracts increased insignificantly (P>0.05). The increases obtained at
600 and 800mg/kg of deionized leaf extracts were not significant (P>0.05), while those of ethylacetate
extracts were significant (P<0.05).The concentration of creatinine, urea and uric acid obtained from
serum of the animals treated with DWL extract was insignificantly higher (P>0.05), while the values
recorded with other extracts were significantly higher (P<0.05) than in control. These results suggest
that deionized water extracts of Newbouldia laevis, as used, may not be toxic to the liver and kidney,
while those of ethylacetate may be. Doses of 200 and 400mg/kg DWL extract may be
hepatoprotective.
Keyword: Liver enzymes; Newbouldia laevis; hepatobilliary toxicity; renal toxicity
INTRODUCTION
The use of plants in the management and
treatment of diseases started with life. In more
recent years with considerable research, it has
been found that many plants do indeed have
J Pharm Chem Biol Sci, September-November 2015; 3(3):367-372
Agbafor & Ezeali
medicinal value and these plants have been
used to make modern medicines which are now
presented by physicians and available for
purchase at the drug stores. Today, traditional
medicine has been brought into focus for
meeting the goals of a wider coverage of
primary healthcare delivery not only in Africa,
but also, to various extents, in all countries of
the world and is the first choice healthcare
treatment for at least 80% of Africans who
suffer from high fever and other common
ailments [1].
In Nigeria many diseases were treated and are
still being treated with medicinal plants with
success. These diseases include malaria,
epilepsy, infantile convulsion diarrhoea,
dysentery, bacterial and fungal infections,
mental illness, asthma, diabetes, worm
infestation, pains and ulcers. Some of the
medicinal plants used in Nigeria include Garcina
kola used for the treatment of asthma; Carica
papaya used as a remedy for hypertension;
Ocinum basilicum as cure for typhoid fevers and
Cola nitida for treatment of piles, etc [2][3][4].
The pharmacological properties of a medicinal
plant stem from the chemical constituents of
the plant. These compounds (phytochemicals,
vitamins and minerals) have the ability to
change physiological states [5].
Newbouldia laevis is a medium sized
angiosperm in the Bignoniaceae family. It is
found tropical Africa and grows to a height of
about 10 meters with a cauliferous habit. It is
ever green, though its leaves turn somewhat
dark purple during the cold seasons. It is
popularly known as the tree of life or fertility
tree in Nigeria. Its local Nigerian names include
Akoko (Yoruba), Aduruku (Hausa), and Ogirisi
(Igbo). The root and leaves are used in the
treatment of diseases such as fever, headache,
convulsion, epilepsy, and manic disorders [6].
Detailed documented information on the
applications of extracts of various parts of
Newbouldia laevis are scarce. Further, like
many medicinal plants, many of the uses and
368
possible adverse effects of the plant by
traditional medicine practitioners have not
been investigated. Hence, this research
investigated possible toxicity of leaf and root
extracts of the plant on hepatic and renal
systems.
MATERIALS AND METHODS
Collection of Leaves and Roots of Newbouldia
laevis
Fresh leaves and roots of Newbouldia laevis
were collected in the month of March, 2015
from Izzi in Abakaliki Local Government of
Ebonyi State. The samples were identified by
Prof. S. C. Onyekwelu of Applied Biology
Department, Ebonyi State University, Abakaliki.
Preparation of Extracts
The samples were washed with distilled water,
air-dried and ground into powder for
extractions.
The methods of extraction used by Agbafor [3]
were adopted, utilizing deionized water and
ethylacetate as solvents. The extracts were
concentrated using rotor evaporator to get gellike dark brown extracts.
Experimental Animals and Handling
Ethical approval for use of animals in research
was given by Ebonyi State University Research
and Ethics Committee.
A total of eighty-five (85) adult male albino rats,
placed in seventeen (17) groups (1-17) of five in
each, were used. Groups 1, 2, 3, and 4 received
oral administration of 200, 400, 600 and
800mg/kg body weight of DWL extract
respectively for twenty one consecutive days,
while group 17 was given normal saline. The
other groups were treated the same way as
summarized below.
Groups 1 - 4 = DWL extract.
Groups 5 – 8 = DWR extract.
Groups 9 – 12 = EAL extract.
Groups 13 – 16 = EAR extract.
Group 17 = normal saline.
J Pharm Chem Biol Sci, September-November 2015; 3(3):367-372
Agbafor & Ezeali
Levels of alanine aminotransferase (ALT),
aspartate aminotransferase (AST), alkaline
phosphatase (ALP) and γ-glutamyl transferase
(GGT) in serum were be used to monitor the
state of hepatic system. Renal toxicity was
evaluated using serum levels of creatinine, urea
and uric acid.
Serum activities of ALT and AST will be
determined according to the method of
Reitman and Frankel [7], as described by
Akubugwo and Agbafor [8]. A manual two-point
kit procedure using p-nitrophenylphosphate
was employed in the determination of serum
alkaline phosphatase activity [9]. The method of
Rosalki and Tarlow [10], described by Tietz [9]
was used to measure the serum activity of GGT.
The method of Jaffe’s reaction described by
Tietz [9] was used to measured serum
creatinine. Serum urea was measured according
to the method described by Fawcett and Scotte
[11]. The method of Brown and Freier [12] was
used for the determination of serum uric acid
concentrations.
STATISTICAL ANALYSIS
Data generated were expressed as mean ± SD.
Statistical significance of difference was
determined using the program SPSS 12 (SPSS,
USA) by performing one-way ANOVA with posthoc comparisons between the control group
and each of the treated groups by Ducan’s
multiple comparison test. A p-value less than
0.05 was considered statistical significant.
RESULTS AND DISCUSSION
Figures 1, 2, 3 and 4 show serum Levels of ALT,
AST, ALP and GGT of the rats after twenty one
days of extracts administration. The activity of
the enzymes decreased insignificantly (P>0.05)
in the groups given 200 and 400mg/kg of DWL
extract (1 and 2). From the figures, the activity
of the enzymes recorded in 600 and 800mg/kg
of DWL extract increased insignificantly
(P>0.05). The increase in activity of these
enzymes all the groups administered DWR
369
extract was not significant (P>0.05).
Administration of EAL extract produced an
insignificant increase (P>0.05) in the enzymes
activity at the dose of 200mg/kg, while the
increase was significant (P<0.05) at the other
doses. The extract doses of 200 and 400mg/kg
of EAR extract recorded an insignificant
increase (P>0.05) in the levels of the enzymes,
while 600 and 800mg/kg gave a significant
increase (P<0.05).
The decrease in activity of ALT, AST, ALP and
GGT produced 200 and 400mg/kg DWL leaf
extract suggests hepatoprotective potential.
This may be due to the reported antioxidant
property of the extracts [13]. Antioxidants
scavenge free radicals and prevent lipid
peroxidation of biomembranes thereby
reducing the leakage of intracellular enzymes
[14].
However, at higher doses of DWL extract and
all doses of other extracts the serum levels of
the enzymes increased linearly in a dosedependent manner. This increase is an
indication of toxicity. Clinical observations and
experimental studies have shown that subtle
changes in the membranes of hepatocytes are
sufficient to allow passage of intracellular
enzymes into the extracellular space. Cell
damage increases membrane permeability,
causing cytosolic enzymes to spill into
circulation [14]. For instance, in liver disease
associated with hepatic necrosis, serum ALT
and AST levels are elevated even before the
clinical signs and symptoms of the disease
appear [14].
Several phytochemicals have been reported to
exhibit hepatotoxicity. Pyrrolizidine alkaloids
are metabolized in the liver to pyrroles which
are very toxic to hepatocytes, causing
hepatocellular death and fibrosis [15]. Nwogu
et al [16] reported the hepatotoxicity of
pyrogallol, a hydrolysable tannin derived from
simple phenolic acids like gallic acid. The levels
of the enzymes obtained in the groups
administered EAL extract were correspondingly
J Pharm Chem Biol Sci, September-November 2015; 3(3):367-372
Agbafor & Ezeali
higher than other groups, suggesting higher
toxicity. This may be due to higher
concentrations of toxic phytochemicals.
The results on examination of kidney function
are presented in figures 5, 6 and 7. The
concentration of creatinine, urea and uric acid
obtained from serum of the animals treated
with DWL extract was insignificantly higher
(P>0.05) than in control. The increase in their
concentration was also insignificant (P>0.05) at
doses of 200, 400 and 600mg/kg of DWR
extract, but significant (P<0.05) at 800mg/kg of
the extract. The groups treated with EAL extract
produced a significant increase (P<0.05) at all
doses except 200mg/kg which was insignificant
(P>0.05). In contrast, the values of these
parameters obtained in the groups given EAR
extract increased insignificantly (P> 0.05) at
doses of 200 and 400mg/kg, and significantly at
600 and 800mg/kg.
The elevations in creatinine, urea and uric acid
concentrations after administration of the
extracts suggest possible renal toxicity.
Measurement of blood urea is presently the
most widely used screening test (with
Fig. 1: ALT activity in the animals after twentyone days of extract administration
370
creatinine) for the evaluation of kidney
function. Renal disorders that lead to high
blood urea levels include chronic nephritis,
polycystic kidney, tubular necrosis and
obstruction of urinary tract [17]. Elevated
serum creatinine levels are observed in
impaired renal function caused by chronic
nephritis, urinary tract obstruction, etc. Uric
acid levels in the serum are elevated in
conditions associated with decreased renal
function [9].
The actual cause of this possible renal toxicity is
still being investigated. However, some of the
chemical constituents of the extracts may have
contributed. Several studies [18][19] have
indicated the possibility that the use of plant
extracts in high doses could lead to toxic injury
to the kidneys, which interfere with renal
tubular functions and induce renal failure. Some
pyrrolizidine alkaloids have been reported to
contribute to renal damage due to
megalocytosis of renal tubular epithelium and
glomerulosclerosis [15]. The EAL extract
showed highest toxicity.
Fig. 2: AST activity in the animals after twentyone days of extract administration.
J Pharm Chem Biol Sci, September-November 2015; 3(3):367-372
Agbafor & Ezeali
Fig. 3: ALP activity in the animals after twentyone days of extract administration
Fig. 4: GGT activity in the animals after twentyone days of extract administration
371
Fig. 6: Serum urea of the animals after twentyone days of extract administration
Fig. 7: Serum uric acid of the animals after
twenty-one days of extract administration
CONCLUSION
The leaf and root extracts may not be as toxic
as many medicinal plants, especially when used
at doses not greater than 400mg/kg. The DWL
extract may be hepatoprotective at doses not
greater than 400mg/kg body weight. Efforts are
in progress in our laboratory to identify the
compounds responsible for these observations.
Fig. 5: Serum creatinine of the animals after
twenty-one days of extract administration
CONFLICT OF INTEREST STATEMENT
The authors declare that they have no
competing interests.
REFERENCES
1. Treben N. Health through God’s Pharmacy,
2nd ed. Australia: Punsthaler Steyr; 1998, p
260 – 264.
2. Sofowora A. The state of Medicinal plants in
Nigeria. 3rd
edition. Nigeria: UI
Press; 1993, p 206 -210.
J Pharm Chem Biol Sci, September-November 2015; 3(3):367-372
Agbafor & Ezeali
3. Agbafor K N. The effect of aqueous and
organic extracts of fresh leaves of Baphia
nitida on tissues acetycholinesterase in
guinea pigs. J Sci Technol 2004; 10: 1-7.
4. Agbafor K N, Akubugwo E I, Ukpabi C F,
Elom S O, Ugwuja E, Nwachukwu N. Wound
healing and antioxidant properties of leaf
extracts of Zapoteca portoricensis. Int J
Bioassay 2014; 3(5): 2066-2069.
5. Oloyede OI. Chemical profile of unripe pulp
of Carica papaya. Pakistan J Nutri 2005; 6:
379 – 381.
6. Ainooson GK, Woode E, Obiri DD, Koffou GA
Antinociceptive Effects of Newbouldia
laevis extract in a rat Model. African J Trad
Complement Alt Med 2009; 5 70: 49-58.
7. Reitman SN, Frankel SA. Colorimetric
method for the determination of glutamate
pyruvate transaminase and glutamate
oxaloacetate transaminase. Am J Clin Pathol
1957; 28(1):56-63.
8. Akubugwo IE, Agbafor KN. Hepatotoxic
evaluation of water and salt from Uburu
and Okposi salt lakes. Estud Biol 2007; 29
(66): 99 - 104.
9. Tietz N. Fundamentals of Clinical Chemistry,
4th ed. Philadelphia: W. B. Saunders; 2000,
p 984 – 1002.
10. Rosalki SB, Tarlow D. Optimized
determination of Y-glutamyltransferase by
reaction-rate analysis. Clin Chem 1974; 20:
1121 – 1124.
372
11. Fawcett JK, Scotte JE. A rapid and precise
method for determination of urea. J Clin
Path 1960; 13:156-159.
12. Brown RA, Freier EF. Serum uric acid levels.
Clinical Biochem 1967’; 1:154-155.
13. Agbafor KN. Ezeali C, Akubugwo EI, Obiudu
IK., Uraku AJ, Ogbanshi ME, Edwin N and
Ugwu OPC. Cardioprotective effect of leaf
and root extracts of Newbouldia laevis
against carbon tetrachloride inducedcardiotoxicity in albino rats. European
Journal of Medicinal Plants 2015; 9(3): 1-7.
14. Burtis CA, Ashwood ER. Tietz Fundamentals
of Clinical Chemistry, 5th ed. India: Elsevier
Pub; 2003.
15. Telcott P. Pyrrolizidine alkaloid poisoning,
in: current therapy in equine medicine. 5th
ed, New Delhi: W.B. Saunders, p 788 – 790.
16. Nwogu LA, Igwe CU, Emejulu AA. Effects of
Landolphia owariensis leaf extracts on the
liver and haemoglobin of albino rats. Afr J
Biotechnol 2008; 2(12): 240 – 242.
17. Renee S. Urea and its blood levels. J
Medical Asso 2001; 3: 23 – 26.
18. Bwititi P, Musabayane CT, Nnachi CF.
effects of Opuntia magagantha on blood
glueose and kidney function in diabetic rats.
J Ethnopharmacol 2000; 63 (3): 247 – 252.
19. Ijeh H, Agbo CA. Body organ weight changes
following administration of aqueous
extracts of Ficus exasperate. Vahl J Anim
Vet Adv 2006; 5: 277 – 279.
Cite this article as:
Agbafor KN, Ezeali C. Effects of Leaf and Root Extracts of Newbouldia laevis on Hepatic and
Renal Systems in Albino Rats. J Pharm Chem Biol Sci 2015; 3(3):367-372
J Pharm Chem Biol Sci, September-November 2015; 3(3):367-372