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NEPHROPROTECTIVE POTENTIALS OF ETHANOL LEAF EXTRACT
OF BALAKAT TREE (
AGAINST GENTAMICIN-INDUCED NEPHROTOXICITY IN MALE
ICR MICE (
Oliver Y. Galang1
, Angelico G. Reyes1 and Lourdes Fatima S. David1
1 Department of Natural Science and Mathematics, Institute of Arts and Sciences, Mabalacat City College, Mabalacat City, Pampanga,
Nephrotoxicity is one of the major causes of renal dysfunction when body exposed to a drug or toxin (Porter and Bennett, 1981). It is con-
sidered the ninth most leading cause of death worldwide (WHO, 2016). In the Philippines, it has been reported as the seventh leading
cause of death due to a lethargic lifestyle which triggers diabetes and hypertension (DOH, 2014). Until now there are no applicable semi-
synthetic drugs created to treat kidney disease; the only efficient treatments for kidney diseases are dialysis and renal replacement.
Renal dysfunction develops due to increased production of reactive oxygen species on the kidneys induced lipid peroxidation which leads to
depletion of endogenous antioxidants (Kim et al., 2006)
Gentamicin is an aminoglycoside antibiotic produced by Micronospora purpurea used to treat nosocomial infections specifically gram–
negative bacteria including pseudomonas, proteus and serratia (Silan et al., 2007). However, it can escalate the blood serum creatinine and
blood urea nitrogen which are the reliable base parameters for renal function (Adelman et al., 1981).Furthermore, there are studies that
show that exogenous antioxidants are needed to protect kidneys against various drugs that have an indication of nephrotoxicity.
Balakat tree (Ziziphus talanai)(Blanco)Merill.) is an endemic plant in the Philippines used as a traditional medicine at San Remigio, Antique,
Philippines against kidney problems including UTI (Anas et al., 2009). Recent studies have shown that Ziziphus talanai possessed hepato-
protective and reproprotective potentials (Reyes et al., 2016).
In addition, preliminary phytochemical screening of ethanol leaf extract of Ziziphus talanai exhibited lower concentrations of triterpenes, al-
kaloids and glycosides; moderate concentration of sterols; and higher concentrations of flavonoids, tannins, and saponins (Bañares, 2016).
There is a paucity of data concerning the property and mechanism of gentamicin on kidneys and as well as nephroprotective potentials of Z.
talanai. The present study evaluated the anecdotal claims and nephroprotective potentials of Z. talanai ethanol leaf extract against gen-
tamicin-induced nephrotoxicity using male ICR mice models.
Collection of leaves
Garlic Soft Gel
Administration of Treatments
T0 Treated with 1ml /20g bw distilled
T- Treated with 0.1ml /20g bw genta-
T+ Treated with 0.2ml/20g bw garlic
Soft Gel Supplement.
T1 Treated with 0.3ml/20g bw Bala-
kat extract after 1 hr 0.1ml gen-
tamicin was administered.
Extraction of kidneys and Blood Serum Histological observations
Figure 1. Statistical Variances of
Figure 2. Means of Blood Urea Nitro-
Figure 3. Histological Observations of
Normal Group (RT) normal Renal tu-
bular, (G) intact Glomerulus.
Figure 4. Histological Observations of
intoxicated– group treated with gen-
Figure 5. Histological Observations of
garlic soft gel supplement exhibited
(G) intact glomerulus, and (RT) normal
Figure 6. Histological viewing treated
with Balakat extract and gentamicin .
(G) intact glomerulus, (RT) normal re-
nal Tubular, moderate (IH) intertubular
This research study was conducted to assess the morphological changes, body weight, kidney weight, blood serum creatinine and
BUN level, and histological observations in terms of glomerulus, renal tubular desquamation, mononuclear cell permeation, inter-
tubular hemorrhage and hyaline cast formation among male ICR mice treated with gentamicin, garlic soft gel and Ziziphus talanai
Male ICR mice treated with ethanol leaf extract of Ziziphus talanai mitigated nephrotoxicity caused by the gentamicin on the kid-
neys. ICR male mice treated with Balakat extract were not irritated, showing red-brown kidney color. Based on the statistical analy-
sis of results, the kidney weight increased significantly, body weight escalated significantly and blood serum analysis such as
creatinine and Blood urea nitrogen were decreased significantly.
Furthermore, the effects of Ziziphus talanai ethanol leaf extract based on the longitudinal histopathology of the kidneys exhibited
intact glomerulus, normal renal tubular and moderate intertubular hemorrhage.
In this study, the observations concerning the effects of gentamicin and Ziziphus talanai ethanol leaf extract were supported by the
related studies. Gentamicin can cause nephrotoxicity due to increased production of free radicals which leads to the depletions of
endogenous antioxidants (Apple et al., 1978). The ethanol leaf extract of Ziziphus talanai has been shown to have histoprotective
potentials against tetracycline-induced hepatoxicity and reprotoxicity (Reyes et al., 2016). The present study provides additional
findings on the nephroprotective potentials of the ethanol leaf extract of Ziziphus talanai against drug-induced nephrotoxicity.
These pharmaceutical potentials are attributable to the bioactive compounds contained in Ziziphus species (Abalaka et al., 2011).
Based on the results, Ziziphus talanai ethanol leaf extract has a potential as nephroprotective agent. Thus, more clinical experi-
ments are recommended that may be undertaken by pharmaceutical companies to create semi-synthetic drugs to cure chronic
kidney disease in the near future.
Based upon the statistical results and histopathological observations, it can be concluded that Ziziphus talanai (Blanco) (Merril.) ethanol leaf ex-
tract exhibited nephroprotective potentials against drug-induced nephrotoxicity due to the bioactive compounds possessed by the Balakat tree
(Ziziphus talanai). Ziziphus related species contain flavonoids and polyphenols which have the capability to scavenge reactive oxygen species
produced by the various drugs including gentamicin which induced nephrotoxicity. Further studies are recommended to elucidate the mechanism
of Z. talanai’s nephroprotective activity.
I would like to express my deepest gratitude to my Alma matter (Mabalacat City College) for funding this activity and to the faculty members spe-
cially; To Mam fhat, Mam Due and Mr, Angelico for the unending support, guidance, motivations and encouragement.
To my Family and to all MCC BSBiology Majors thank you all.
1. Abalaka, M. E, Mann, A and Adeyemo S. O (2011). Studies on in-vitro antioxi-
dant and free radicalscavenging potential and phytochemical screening of
leaves of Ziziphus mauritiana L. and Ziziphus spinachristi L. compared
with Ascorbic acid. Journal of Medical Genetics and Genomics Vol. 3(2),
pp. 28 – 34.
2. Anas, Andrea Roxanne J. et al. (2009).Anti-Mycobacterium phlei activity of
the bark of Ziziphus talanai (Blanco) Merrill. Philipp Agric Scientist, 92(4):
3. Appel, G.B. and Neu , H.C.: Gentamicin in 1978. Ann. Intern. Med. 89:528-
4. Porter GA and Bennett WM. Nephrotoxic acute renal failure due to common
drugs. American journal of Physiology, 1981; 241(7): F1-F8.
5. Reyes, Angelico G. Renato A. Dela Peña, Jr , Lourdes Fatima I. Sula, An-
gelo B. Bañares (2016 ). Histoprotective potentials of ethanol leaf extract
of balakat tree (Ziziphus talanai(blanco) merr.) against tertracycline- in-
duced hepatoxicity and reprotoxicity in male mice (Mus muculus). Interna-
tonal Journal of Pharmacology and Toxicology .4(2) 96-104
Air-drying of leaves