Ogbe et al.

Clinical Phytoscience (2020) 6:4

https://doi.org/10.1186/s40816-019-0149-4

ORIGINAL CONTRIBUTION Open Access

Protective effect of aqueous extract of

Lophira lanceolata leaf against cisplatin-
induced hepatorenal injuries and
dyslipidemia in Wistar rats
Raphael John Ogbe, Samuel Peter Agbese and Adakole Hyacinth Abu*

Abstract
Background: Hepatorenal injuries and dyslipidemia are common global health challenges but medicinal plant
extracts may have potential to prevent them. Thus, this study evaluated the protective effect of aqueous extract of
Lophira lanceolata leaf (LLE) against cisplatin-induced hepatorenal injuries and dyslipidemia in albino Wistar rats.
Methods: Thirty rats were randomly divided into 6 groups of 5 rats each. Group I rats received distilled water and
served as control, group II rats were given 5 mg/kg cisplatin (CIS) intraperitoneally, groups III and IV rats were
treated with 200 and 400 mg/kg LLE respectively for 26 days by oral gavages while groups V and VI rats were
treated with 200 and 400 mg/kg LLE respectively, followed by CIS on the 21st day as in group II. About 24 h after
treatment, blood was collected from the rats; then serum was separated and used for estimations of biochemical
parameters. The kidney and liver of rats were removed, rinsed in normal saline, stored in 10% formalin and used for
histological analyses.
Results: The biomarkers of hepatic (Aminotransferases, Alkaline phosphatase and Bilirubin) and renal (urea and
creatinine) injuries, and dyslipidemia (Total cholesterol, triglycerides and LDL-cholesterol) significantly (p < 0.05)
increased in the rats exclusively exposed to cisplatin when compared with normal control. However, treatment of
cisplatin-exposed rats with 200 and 400 mg/kg LLE significantly (p < 0.05) reduced the levels of these biomarkers of
hepatorenal injuries and dyslipidemia when compared with cisplatin control. Photomicrographs showed
pathological signs in the liver and kidney of rats exclusively exposed to cisplatin, but there was moderate
protection of these tissues in the rats treated with LLE and cisplatin.
Conclusion: The current findings have shown that Lophira lanceolata leaf extract may provide moderate protection
against cisplatin-induced hepatorenal injuries and dyslipidemia in albino Wistar rats.
Keywords: Hepatotoxicity, Histopathology, Hyperlipidemia, Nephrotoxicity

Introduction homeostasis and excretion of drugs, so it is exposed to

The liver plays a central role in transforming and detoxi- drug-induced injuries [3]. The incidence of drug-
fying chemicals, so it is susceptible to their toxic effects induced kidney injury has increased due to increasing
[1]. Drug-induced liver injury is a cause of acute and use of drugs, as they are readily available as over-the-
chronic liver diseases in animals and humans. Recent counter medicines, especially the non-steroidal anti-
epidemiological data suggests that approximately 20 new inflammatory drugs (NSAIDs) and antibiotics [4]. About
cases of drug-induced liver injury (DILI) per 100,000 20% of nephrotoxicity is induced by drugs but medica-
persons occur annually [2]. The Kidney plays a role in tion of the elderly increases this incidence up to 60% as
the average lifespan increases [5]. Dyslipidemia is a con-
* Correspondence: adakoleabu1@gmail.com dition characterized by alteration in the plasma concen-
Department of Veterinary Physiology and Biochemistry, College of Veterinary trations of lipids (triglycerides and total cholesterols)
Medicine, Federal University of Agriculture, Makurdi P. M. B. 2373, Nigeria

© The Author(s). 2020 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made.
Ogbe et al. Clinical Phytoscience (2020) 6:4 Page 2 of 11

and the blood transporting lipoproteins (LDL-choles- h, with frequent agitation after every 2 h. The liquid
terol, HDL-cholesterol and VLDL-cholesterol). Hyperlip- mixture was filtered with Whatman no.1 filter papers.
idemia is a common form of dyslipidemia in animals [6]. The filtrate was concentrated and dried by evaporation
Cisplatin, chemically known as cis-diamine dichloro- at 40 °C using a water-bath. The dried extract was
platinum II (CDDP), with chemical formula cis-[Pt weighed to determine the yield and stored in refrigerator
(NH3)2(Cl)2], is a platinum-based chemotherapeutic until needed.
agent used for treatment of various types of cancers [7].
It is associated with several toxic side effects including Experimental animals and management
nephrotoxicity, hepatotoxicity and oxidative stress injur- Adult male albino Wistar rats (Rattus norvegicus) weigh-
ies. The adverse effects of cisplatin occur by increased ing 150–230 g, were purchased from the Animal house
production of reactive oxygen species (ROS) and de- of College of Health Sciences, Benue State University,
creased production of antioxidants, though the forma- Makurdi, Nigeria. The rats were allowed to stabilize and
tion of ROS depends on the concentration and duration acclimatize for at least 2 weeks before being used for the
of exposure to the drug [8]. experiments. They were kept in plastic cages in well ven-
Herbal medicine is increasingly getting attention as a tilated room, fed with standard animal feeds, produced
useful tool in the management of several disorders. by Grand Cereals and Oil Mills Ltd., Jos, and water ad
Lophira lanceolata known as “Iron wood” (English) is a libitum. The animals were handled with care, according
tree that grows up to 12 m high, with a narrow crown, to the principles and standard protocols for the use of
deep root, less scaly bark and contains no latex [9]. It laboratory animals for experiments.
belongs to the Family: Ochnaceae, widely distributed in
the West and Central African countries including Animal grouping and treatments
Nigeria and Cote d’Ivoire [10]. In Nigeria, it is called The rats were divided into six groups of five animals per
“Okopia” (Igbo), “Ikponhon” (Yoruba), “Okopi” (Idoma) group. Group I rats received only distilled water for 26
and “Namijin kadanya” (Hausa) [11]. It is used for the days by oral gavages and served as normal control. Group
treatment of dermatosis, toothache and muscular tired- II rats were given distilled water for 26 days by oral ga-
ness [9]. It is used by many communities in northern vages and administered 5 mg/kg body weight cisplatin by
Nigeria for the treatment of epilepsy and erectile dys- intraperitoneal (i.p.) route on the 21st day. Groups III and
function [10]. Previous scientific studies on the leaf ex- IV rats received 200 and 400 mg/kg b. wt. Lophira lanceo-
tract of this plant revealed antiplasmodial [12] and lata extract (LLE) respectively for 26 days by oral gavages.
antibacterial [13] activities. However, there may be pau- Group V and VI rats received 200 and 400 mg/kg b. wt.
city of scientific report on the effect of L. lanceolata leaf LLE respectively for 26 days while cisplatin was adminis-
extract on drug-induced hepatorenal disorders and dys- tered on the 21st day as in group II. Body weights of the
lipidemia in animals. Thus, this study was initiated to in- rats were recorded at the initial week, then second, third
vestigate the protective effect of aqueous extract of L. and last weeks of the experiment.
lanceolata leaf against cisplatin-induced hepatorenal in-
juries and dyslipidemia in albino Wistar rats. Collection and preparation of blood and tissues
About 24 h after treatments, rats were sacrificed under
Materials and methods chloroform anaesthesia. Blood was collected from rats
Drug used for induction of hepatorenal injuries and by intra-cardiac puncture and dispensed into plain steril-
dyslipidemia ized tubes. The blood was allowed to stand on the bench
Cisplatin was the drug of choice for induction of dyslip- for at least 2 h, spun with a centrifuge at 3000 rpm for
idemia, liver and kidney injuries in rats. It is an inject- 10 min, then serum was separated with clean Pasteur
able liquid manufactured by Kwality Pharmaceuticals pipette and stored frozen until used for biochemical ana-
Pvt. Ltd., Nagkalan, Majitha Road, Amritsar, India. lyses. The liver and kidney of rats were excised, rinsed in
normal saline, weighed and preserved in 10% formalin
Plant collection and preparation for histological analyses.
The fresh leaves of L. lanceolata were collected from
North Bank - Gbajimba road, in Makurdi, Benue State, Biochemical assays
Nigeria, and identified by a plant taxonomist. The plant The biochemical parameters, which are markers of hepa-
leaves were air-dried at room temperature for 2 weeks, torenal injuries and dyslipidemia, were estimated using
pulverized using a grinding machine, and sieved to ob- assay kits according to the manufacturer’s procedures.
tain a fine powder. A 200 g of pulverized sample was High density lipoprotein (HDL) cholesterol concentra-
macerated in 4000 ml of distilled water in a conical flask tion was determined by the procedure described in the
and covered. This was kept at room temperature for 48 reagent kits instruction manual (AGAPPE Diagnostics
Ogbe et al. Clinical Phytoscience (2020) 6:4 Page 3 of 11

Ltd., Switzerland). Total cholesterol (TC) and triglycer- total bilirubin levels of cisplatin-exposed rats when com-
ide (TRG) concentrations were determined by the pared with the normal control. There was no significant
methods given in the reagent kits instruction manuals (p > 0.05) difference in the levels of total proteins and al-
(TECO Diagnostics Ltd., USA). Alanine aminotransfer- bumin of rats in all the treatment groups compared with
ase (ALT) and aspartate aminotransferase (AST) activ- normal control. However, treatment with aqueous ex-
ities were determined by IFCC methods, using the kits tract of L. lanceolata leaf significantly (p < 0.05) reduced
instruction manual (AGAPPE Diagnostics Ltd). Alkaline the levels of ALT, AST and total bilirubin of cisplatin-
phosphatase activity was determined by the method de- exposed rats when compared with the cisplatin control
scribed in the assay kit instruction manual (TECO diagnos- (Table 1).
tics Ltd., USA). Serum bilirubin levels were determined by
the modified TAB method described in the assay kits man- Effect of Lophira lanceolata leaf extract on lipid profile
ual (AGAPPE Diagnostic Ltd). The creatinine and total and biomarkers of renal injuries in rats
protein concentrations were determined by the methods There was a significant (p < 0.05) increase in the levels of
described in assay kits instruction manuals (Randox T-cholesterol, triglycerides and LDL-cholesterol of
Laboratories Ltd., UK). The urea and albumin concentra- cisplatin-exposed rats when compared with the normal
tions were estimated by the methods described in assay kits control. There was no significant (p > 0.05) difference in
manuals (AGAPPE Diagnostics Ltd). the levels of HDL-cholesterol of rats in all the treatment
The atherogenic/dyslipidemic indices were calculated groups. However, treatment with 200 and 400 mg/kg b.
using the equations, according to previously described wt. aqueous extract of L. lanceolata leaf significantly
methods [14, 15]. (p < 0.05) reduced the levels of T-cholesterol, triglycer-
ides and LDL-cholesterol of cisplatin-exposed rats when
1. Cardiac risk ratio (CRR) = Total cholesterol/HDL compared with the cisplatin control (Table 2). There
cholesterol was a significant (p < 0.05) increase in the levels of urea
2. Atherogenic coefficient (AC) = Total cholesterol – and creatinine of cisplatin-exposed rats when compared
HDL cholesterol/HDL cholesterol with normal control. However, treatment with 200 and
3. Atherogenic index of plasma (AIP) = Log 400 mg/kg b. wt. aqueous extract of L. lanceolata leaf
(Triglycerides/HDL cholesterol) significantly (p < 0.05) reduced the levels of urea and cre-
4. Classical ratio (CR) = LDL cholesterol/HDL atinine of cisplatin-exposed rats when compared with
cholesterol cisplatin control (Table 2).

Histological analyses of liver and kidney tissues Effect of Lophira lanceolata leaf extract on percentage
Histopathological examination was carried out according to organ to body weight ratio of rats
a previously described method [16]. The liver and kidney There was a significant (p < 0.05) reduction in the
tissues were fixed in 10% formalin, dehydrated, embedded weights of liver, kidney, ratio of liver to body weight and
in paraffin, sectioned and stained with hematoxylin and kidney to body weight of cisplatin-exposed rats when
eosin. The stained glass slides were viewed under light compared with normal control. The treatment of
microscope at × 10 and 40 objective lenses. cisplatin-exposed rats with 200 and 400 mg/kg LLE sig-
nificantly (p < 0.05) increased their ratio of liver to body
Statistical analysis weight and kidney to body weight when compared with
The statistical package for the social sciences (SPSS ver- cisplatin control. There was also significant (p < 0.05) in-
sion 21.0) produced by SPSS Inc., Chicago, USA, was crease in the weights of liver and kidney of cisplatin-
used for statistical analysis. Data were presented as exposed rats treated with 200 mg/kg LLE when com-
mean ± standard error of mean (SEM), with n = 5. The pared with cisplatin control. However, there was no sig-
data were analyzed by one-way analysis of variance nificant (p > 0.05) difference in the weights of liver and
(ANOVA), followed by least significant difference kidney of rats given 200 and 400 mg/kg LLE only when
(LSD) as Post Hoc test for multiple comparisons. Mean dif- compared with the normal control (Table 3).
ferences were considered statistically significant at p < 0.05.
Effect of Lophira lanceolata leaf extract on atherogenic/
Results dyslipidemic indices in rats
Effect of Lophira lanceolata leaf extract on biomarkers of There were increases in cardiac risk ratio (CRR), athero-
hepatic injuries in rats genic coefficient (AC), classical ratio (CR) and athero-
There was a significant (p < 0.05) elevation in the activ- genic index of plasma (AIP) of cisplatin-exposed rats
ities of serum alanine aminotransferase (ALT), aspartate when compared with the normal control. The rats
aminotransferase (AST), alkaline phosphatase (ALP) and treated with 200 mg/kg b. wt. showed slight increases in
Ogbe et al. Clinical Phytoscience (2020) 6:4 Page 4 of 11

Table 1 Effect of Lophira lanceolata leaf extract on biomarkers of hepatic injuries in rats administered cisplatin
Treatment groups Serum biomarkers of hepatic injuries
T-Protein (g/dL) Albumin (g/dL) ALT (U/L) AST (U/L) ALP (U/L) T-Bilirubin (mg/dL)
I. Normal control 6.68 ± 0.69 3.00 ± 0.13 30.12 ± 3.46 27.56 ± 5.17 22.32 ± 6.59 0.62 ± 0.12
II. 5 mg/kg Cisplatin 6.42 ± 0.70 3.50 ± 0.32 178.0 ± 38.5a 73.44 ± 2.75a 48.04 ± 3.09a 3.78 ± 1.15a
b b
III. 200 mg/kg LLE 6.48 ± 0.41 3.12 ± 0.26 33.72 ± 5.98 37.14 ± 9.58 35.32 ± 5.87 0.56 ± 0.09b
IV. 400 mg/kg LLE 5.56 ± 0.37 2.98 ± 0.16 31.50 ± 2.83b 30.22 ± 3.26b 39.36 ± 10.1 0.48 ± 0.17b
b b
V. 200 mg/kg LLE + CIS 7.28 ± 0.88 3.12 ± 0.16 44.08 ± 8.18 42.56 ± 4.49 34.60 ± 6.28 0.40 ± 0.14b
VI. 400 mg/kg LLE + CIS 6.24 ± 0.42 2.90 ± 0.20 66.40 ± 6.65b 34.16 ± 2.82b 34.40 ± 5.36 1.86 ± 0.10b
Values are Mean ± SEM, n = 5; LLE - Lophira lanceolata leaf extract, CIS - Cisplatin
a
significantly different from normal control (p < 0.05)
b
significantly different from cisplatin control (p < 0.05)

CRR, AC, and CR while those treated with 400 mg/kg b. wt. The photomicrographs of liver and kidney sections of
showed slight decreases in CRR, AC and AIP when com- rat treated with 200 mg/kg LLE showed mild hepatorenal
pared with normal control. However, the treatment of injuries. There was mild congestion of interstitial spaces
cisplatin-exposed rats with 200 and 400 mg/kg b. wt. LLE and foci of interstitial nephritis. There was infiltration of
decreased their CRR, AC, CR and AIP when compared with hepatic cords by few inflammatory cells and foci of mild
cisplatin control. Though, the decreases in CRR, AC, CR coagulative necrosis (Figs. 3a and b). The photomicro-
and AIP values were greater by treatment of cisplatin- graphs of liver and kidney sections of rat treated with
exposed rats with 200 mg/kg than 400 mg/kg LLE (Table 4). 400 mg/kg LLE showed mild hepatorenal injuries. There
was mild congestion of renal interstitial spaces, and
some glomeruli were shrunken with dilated capsular
Histopathological findings in liver and kidney of rats spaces. There was infiltration of portal triad and hepatic
treated with L. lanceolata extract cords by inflammatory cells, and mild congestion of liver
The photomicrographs of liver and kidney sections of rat sinusoids (Figs. 4a and b).
given only distilled water appeared normal, without signs The photomicrographs of liver and kidney sections of
of degenerative changes. The hepatocytes and liver sinu- rat treated with 200 mg/kg LLE and exposed to 5 mg/kg
soids appeared normal while the glomerulus, Bowman’s CIS showed mild hepatorenal injuries. There were few
capsule and renal tubules also appeared normal (Figs. 1a shrunken glomeruli, infiltration of renal tubules by in-
and b). The photomicrographs of liver and kidney sections flammatory cells and congestion of interstitial spaces,
of a rat exposed to 5 mg/kg b.wt. CIS showed signs of showing signs of mild interstitial nephritis. There was
hepatorenal injuries. There was sinusoidal dilatation, vas- congestion of few sinusoids in the liver, which was mild
cular congestion, infiltration of portal triad and stroma of when compared with the CIS control (Figs. 5a and b).
liver by inflammatory cells, and foci of coagulative necro- The photomicrographs of liver and kidney sections of
sis. The kidney tissues showed capsular dilation, distorted rat treated with 400 mg/kg LLE and exposed to 5 mg/kg
glomeruli, and congestion of interstitial spaces, showing CIS showed mild hepatorenal injuries and signs of re-
signs of interstitial nephritis (Figs. 2a and b). covery. There were pockets of hemorrhage seen in the

Table 2 Effect of Lophira lanceolata leaf extract on lipid profile and biomarkers of renal injuries in rats administered cisplatin
Treatment groups Serum lipid profile and biomarkers of renal injuries
Total Chol. (mg/dL) TRG (mg/dL) HDL-Chol. (mg/dL) LDL-Chol. (mg/dL) Urea (mg/dL) Creatinine (mg/dL)
I. Normal control 116.6 ± 3.1 129.2 ± 5.1 36.82 ± 3.09 46.84 ± 3.15 27.46 ± 4.68 0.44 ± 0.17
II. 5 mg/kg Cisplatin 194.8 ± 24.0a 149.5 ± 2.9a 36.68 ± 3.20 133.5 ± 17.4a 65.14 ± 6.96a 1.92 ± 0.39a
b b b b
III. 200 mg/kg LLE 127.2 ± 11.9 117.7 ± 4.2 37.00 ± 3.84 58.78 ± 8.20 32.34 ± 3.71 1.06 ± 0.24b
IV. 400 mg/kg LLE 94.2 ± 8.01b 136.1 ± 1.9 40.40 ± 1.30 56.22 ± 6.10b 33.08 ± 4.33b 0.48 ± 0.07b
b b b
V. CIS + 200 mg/kg LLE 109.3 ± 5.12 146.6 ± 14.6 41.44 ± 2.24 31.72 ± 5.48 30.28 ± 1.10 0.46 ± 0.09b
VI. CIS + 400 mg/kg LLE 129.8 ± 0.82b 128.8 ± 1.9b 35.54 ± 4.73 80.92 ± 5.27ab 38.08 ± 5.67b 0.42 ± 0.19b
Values are Mean ± SEM, n = 5; LLE - Lophira lanceolata leaf extract, CIS - Cisplatin
a
significantly different from normal control (p < 0.05)
b
significantly different from cisplatin control (p < 0.05)
Ogbe et al. Clinical Phytoscience (2020) 6:4 Page 5 of 11

Table 3 Effect of Lophira lanceolata leaf extract on percentage of organ to body weight ratio of rats administered cisplatin
Treatment groups Body weight and percentage of organ to body weight ratio
Body weights(g) Liver weights(g) Liver/Body weights (%) Kidney weights(g) Kidney/Body weights (%)
I. Normal control 195.0 ± 10.37 5.66 ± 0.28 2.9 ± 0.20 0.65 ± 0.05 0.33 ± 0.01
II. 5 mg CIS 233.8 ± 11.08a 4.25 ± 0.08a 1.8 ± 0.10a 0.49 ± 0.01a 0.21 ± 0.009a
b b b
III. 200 mg LLE 195.4 ± 5.27 5.91 ± 0.09 3.02 ± 0.07 0.58 ± 0.01 0.30 ± 0.006b
IV. 400 mg LLE 198.6 ± 6.21 6.66 ± 0.27b 3.35 ± 0.16b 0.60 ± 0.01b 0.30 ± 0.008b
b b b
V. 200 mg LLE + CIS 209.0 ± 7.38 5.59 ± 0.38 2.67 ± 0.14 0.72 ± 0.01 0.34 ± 0.01b
VI. 400 mg LLE + CIS 197.4 ± 3.78b 5.00 ± 0.27 2.53 ± 0.09b 0.76 ± 0.01ab 0.39 ± 0.02b
Values are Means ± SEM, n = 5; LLE - Lophira lanceolata leaf extract, CIS - Cisplatin
a
significantly different from the normal control (p < 0.05)
b
significantly different from the cisplatin control (p < 0.05)

interstitial spaces and signs of recovery from renal injur- impaired hepatic excretory function, a sign of liver dys-
ies. There was mild hemorrhage and infiltration of portal function, attributed to the adverse effect of cisplatin.
triad of the liver by inflammatory cells, which was less These findings are in agreement with El-Hosseiny et al.
severe when compared with CIS control (Figs. 6a and b). [2] who found significant elevation in the levels of total
bilirubin, after exposure of rats to a drug. Bilirubin is the
Discussion excretory product formed by catabolism of heme in
The alterations of alanine aminotransferase (ALT) and hemoglobin, thus hyperbilirubinemia is usually caused
aspartate aminotransferase (AST) activities are biochem- by excessive destruction of heme and blockage of biliary
ical indicators of hepatocellular injuries [17]. The signifi- tract; which results in severe inhibition of conjugation
cant increase in serum AST and ALT activities, after reaction and release of unconjugated bilirubin from
exposure of rats to cisplatin may indicate liver injuries, damaged and dead hepatocytes [18]. The significant re-
attributed to the adverse effect of this drug. Previous duction of AST, ALT, ALP and bilirubin levels after
studies have shown that increases in serum AST and treatment of cisplatin-exposed rats with LLE may indi-
ALT activities reflect hepatocellular membrane damage cate a protective effect of this plant extract against
and leakage [2, 17]. The significant increase in alkaline cisplatin-induced hepatic injuries. The LLE may have
phosphatase (ALP) activity may also indicate liver im- stabilized hepatocyte membrane integrity against cis-
pairments and cholestasis, due to the adverse effect of platin lethality, thereby preventing elevation of these bio-
cisplatin. ALP activity increases as a result of biliary ob- markers of hepatic injuries in rats. Some plant extracts
structions or cholestasis, thus it is a biomarker of hepa- have earlier been reported to produce protective effects
tobiliary disease [2]. against drug-induced liver injuries in animals by reduc-
The alteration in serum bilirubin levels is a biochem- tion of ALT, AST, ALP and bilirubin levels [2, 19].
ical indicator of the changes in morphological integrity Creatinine and urea levels are used as biochemical
of hepatobiliary tract, as a sign of liver damage [17–19]. markers of kidney injuries, thus elevated levels of these
Elevated levels of bilirubins may be due to excessive markers may indicate kidney dysfunction, though the
haemolysis or biliary tract obstruction [18]. The signifi- test for creatinine is a better indicator than urea [20].
cant increase in serum total bilirubin may indicate Creatinine is a non-protein nitrogenous substance

Table 4 Effect of Lophira lanceolata leaf extract on atherogenic/dyslipidemic indices in rats administered cisplatin
Treatment groups Atherogenic/dyslipidemic indices of rats
Cardiac risk ratio (CRR) Atherogenic coefficient (AC) Atherogenic index (AIP) Classical ratio (CR)
Normal control 3.17 2.17 0.55 1.27
a a a
Cisplatin control 5.31 (67) 4.31 (98) 0.61 (11) 3.64 (186)a
a a a
200 mg/kg LLE only 3.44 (9) 2.44 (12) 0.50 (9) 1.59 (25)a
400 mg/kg LLE only 2.33 (27)a 1.33 (39)a 0.53 (4)a 1.39 (9)a
b b b
200 mg/kg LLE + CIS 2.64 (50) 1.64 (62) 0.55 (10) 0.76 (79)b
400 mg/kg LLE + CIS 3.65 (31)b 2.65 (39)b 0.56 (8)b 2.27 (38)b
Values are means of 5 rats; Values in parenthesis are percentage changes (%), CIS - Cisplatin, LLE - Lophira lanceolata leaf extract
a
percentage changes when compared with normal control
b
percentage decrease when compared with cisplatin control
Ogbe et al. Clinical Phytoscience (2020) 6:4 Page 6 of 11

Fig. 1 Photomicrographs of liver and kidney of rat given distilled water only (× 100, H & E stain); Morphology of liver tissue (a) showing normal
hepatocytes, without any change in the tissue structural architecture, and kidney tissue (b) with normal glomeruli, Bowman’s capsules and
renal tubules

formed from creatine and phosphocreatine during rats with LLE and cisplatin may suggest that this plant
muscle metabolism but excreted by glomerular filtration. extract has protective effect against cisplatin-induced
Just like urea, the rate of excretion of creatinine is influ- kidney injuries in animals. The capacity of this plant ex-
enced by glomerular filtration rate (GFR), so any abnor- tract to decrease urea and creatinine levels in rats may
mality that decreases GFR will result in an increased be attributed to its ability to protect the nephrons and
serum creatinine [21, 22]. The significant increase in increase GFR, as a result of its phytochemicals [21]. Pre-
urea and creatinine levels after exposure of rats to cis- vious studies have shown that certain plant extracts have
platin may be an indication of kidney impairments, due protective effects against cisplatin-induced kidney injur-
to the adverse effect of this drug. These findings suggest ies in animals by reduction of urea and creatinine levels
that there may be reduction in GFR of the animals and [1, 20, 22].
are in agreement with the findings of Kim et al. [1] and The liver plays a major role in controlling plasma
Rajakrishnan et al. [20] who reported that the alterations levels of cholesterol, thus when there is drug-induced
induced in kidneys by cisplatin were characterized by el- liver impairments, there will be elevated levels of serum
evated levels of their injury markers. The significant re- total cholesterol (TC) and LDL-cholesterol [17]. The sig-
duction of urea and creatinine levels after treatment of nificant increase in serum levels of total cholesterol

Fig. 2 Photomicrographs of liver and kidney of rat administered 5 mg/kg cisplatin (× 100, H & E stain); Morphology of liver tissue (a) shows
damaged hepatocytes, sinusoidal dilatation, hemorrhage in portal triad, vascular congestion, infiltration of portal triad and stroma of liver by
inflammatory cells, and foci of coagulative necrosis. The kidney tissue (b) shows congestion of interstitial spaces with presence of casts, capsular
dilation, shrunken and distorted glomeruli
Ogbe et al. Clinical Phytoscience (2020) 6:4 Page 7 of 11

Fig. 3 Photomicrographs of liver and kidney of rat treated with 200 mg/kg L. lanceolata extract (× 100, H & E stain); Morphology of liver tissue (a)
shows infiltration of hepatic cords by few inflammatory cells and small foci of coagulative necrosis. The kidney tissue (b) shows mild congestion
of interstitial spaces, shrunken glomeruli and foci of interstitial nephritis

(TC), triglycerides (TG) and LDL-cholesterol after expos- and useful for assessing response to treatment instead of
ure of rats to cisplatin may be attributed to the adverse ef- the classical ratio [26, 27]. It was then found that athero-
fect of this drug, leading to hepatocellular dysfunction and genic index of plasma (AIP), as a marker of lipoprotein
impaired lipid metabolism, which are in agreement with particle size, has predictive value for CVD risk than indi-
the findings of Akindele et al. [23]. It has been shown that vidual lipids and/or TC/HDL-C ratio [26, 28]. Thus,
elevated levels of TC, LDL-C and reduced level of HDL-C current scientific evidence indicates that AIP is a strong
are risk factors for cardiovascular diseases [24]. The liver marker which can predict the risk of atherosclerosis and
synthesizes triglycerides and incorporates them into very CVD [26]. The marked increase in the values of cardiac
low density lipoprotein (VLDL) cholesterol for transport risk ratio (CRR), atherogenic coefficient (AC), and clas-
into peripheral tissues, thus when there is injury to the sical ratio (CR) but minimal increase in AIP of cisplatin-
liver its capacity to synthesize VLDL-C is impaired, lead- exposed rats may indicate a moderate risk of athero-
ing to elevated levels of TG [25]. sclerosis and CVD. These findings are in agreement with
Atherogenic dyslipidemia, characterized by high LDL- previous studies, which showed that dyslipidemic and
C/HDL-C ratio and elevated TG, is associated with high atherogenic indices increase with increasing risk of CVD
cardiovascular risk [26]. Several clinical studies tried to and vice versa [26–29].
find a better marker of atherogenic dyslipidemia, which The significant reduction in the levels of TC, LDL-C
can predict the risk of cardiovascular diseases (CVD) and TG by treatment of cisplatin-exposed rats with 200

Fig. 4 Photomicrographs of liver and kidney of rat treated with 400 mg/kg L. lanceolata extract (× 100, H & E stain); Morphology of liver tissue (a)
shows infiltration of portal triad and hepatic cords by inflammatory cells, and vascular congestion. The kidney tissue (b) shows congestion of
interstitial spaces, shrunken and distorted glomeruli
Ogbe et al. Clinical Phytoscience (2020) 6:4 Page 8 of 11

Fig. 5 Photomicrograph of liver and kidney of rat treated with 200 mg/kg L. lanceolata leaf extract and 5 mg/kg Cisplatin (× 100, H & E stain);
Morphology of liver tissue (a) shows hemorrhage in the portal triad, few inflammatory cells and pyknotic hepatocytes. The kidney tissue (b)
shows shrunken glomeruli, congestion of interstitial spaces and infiltration by inflammatory cells, showing signs of mild interstitial nephritis

and 400 mg/kg LLE may indicate that the plant extract The weight of animal’s organ in relation to body
has protective effect against cisplatin-induced hyper- weight is used to assess the level of tissue damage in ani-
lipidemia in rats, when given at the appropriate dose. mals [30]. It is widely accepted that alteration in the
These findings are in agreement with Akindele et al. organ to body weight ratio of diseased animal is a useful
[23] who found that plant extract has ameliorative ef- indicator of tissue damage [31]. The significant reduc-
fect on ethanol-induced elevated levels of cholesterol, tion in the values of liver and kidney to body weight ra-
TG and LDL-C in rats. The decrease in the values of tios after exposure of rats to cisplatin may indicate
CRR, AC, AIP and CR by treatment of cisplatin- tissues degeneration and necrosis, attributed to the ad-
exposed rats with 200 and 400 mg/kg LLE may sug- verse effects of this drug. These findings are in agree-
gest a reduction in the risk of atherosclerosis and ment with Adebayo et al. [30] who found significant
CVD, though the effect of low dose of extract was reduction in kidney to body weight ratio after exposure
greater than the high dose. These findings are in of rats to a toxin, and Olaniyan et al. [31] who reported
agreement with Akindele et al. [23] who found that that the alteration in organ to body weight ratio may be
the values of CRR, AC and AIP were considerably re- as a result of organ damage.
duced in ethanol-exposed rats by treatment with a The significant increase in the values of liver and kid-
medicinal plant extract. ney to body weight ratios by treatment of cisplatin-

Fig. 6 Photomicrographs of liver and kidney of rat treated with 400 mg/kg L. lanceolata leaf extract and 5 mg/kg Cisplatin (× 100, H & E stain).
Morphology of liver tissue (a) shows mild congestion of the portal triad and infiltration of liver sinusoids by inflammatory cells and several
pyknotic hepatocytes; The kidney tissue (b) shows shrunken and distorted glomeruli, mild hemorrhage and congestion in the interstitial spaces,
showing signs of mild interstitial nephritis
Ogbe et al. Clinical Phytoscience (2020) 6:4 Page 9 of 11

exposed rats with 200 and 400 mg/kg LLE, may suggest apoptotic cell death. The apoptotic bodies once released are
that this plant extract has protective effect against rapidly recognized by neighboring cells or professional
cisplatin-induced liver and kidney injuries in rats. These phagocytes and are generally disposed without induction of
findings are in agreement with Adejor et al. [32] who inflammation [36, 37].
found that the protective effect of plant extract pre- The marked decrease in number of inflammatory cells,
vented significant alteration in organ to body weight ra- reduced degenerative changes in kidney and liver tissues,
tio of animals. Thus, the prevention of marked decrease by treatment of cisplatin-exposed rats with LLE, sup-
in liver and kidney to body weight ratios in the current ported the biochemical findings that this plant extract
study, may suggest that LLE protected the liver and kid- may have protective effect against cisplatin-induced hep-
ney of rats against cisplatin-induced injuries and atic and renal injuries in rats. Previous studies have
necrosis. shown that plant extracts could prevent pathological
The liver and kidney photomicrographs are commonly changes in liver and kidney tissues of animals and some-
used to evaluate the extent of tissue damage, by the times may reverse pathological changes already induced
microscopic study of their structural architectures. Thus, by drugs that are toxic to animal tissues [19, 32, 33].
histopathological findings in tissue sections of animals Though, no functional data was determined to investi-
are commonly used to validate the biochemical findings gate the mechanism of protection against drug-induced
of a study [33]. The histopathological changes observed tissue damage, the protective effect of LLE in the current
in the photomicrographs of cisplatin-exposed rats agreed study may be attributed to the presence of phytochemi-
with the biochemical findings of this study. The presence cals which can scavenge ROS and free radicals generated
of inflammatory cells and alterations in structural archi- by the drug, thereby preventing the oxidative damage of
tectures of liver and kidney tissues of cisplatin-exposed hepatorenal tissues in rats [23]. The changes in struc-
rats are signs of pathology; indicating degenerative tural architectures of liver and kidney tissues of rats after
changes in hepatorenal tissues, by apoptotic and necrotic treatment with LLE only, may suggest that this plant ex-
processes leading to cell death. These findings are in tract may have adverse effect on animal tissues, when
agreement with previous studies by Alqasoumi [19] and given at higher doses and for long duration. These find-
Al-Attar et al. [33] who found pathological changes in ings are in agreement with Etuk and Muhammad [11]
liver and kidney structures after exposure of animals to who found signs of injuries in tissues of male rats after
drugs; with features such as sinusoidal dilation, paren- their treatment with similar plant extract.
chyma inflammation and vascular congestion in liver,
glomerular and tubular modifications in nephrons. Apop- Conclusion
totic and necrotic cell death are important in the patho- The findings of this study have shown that aqueous ex-
genesis of acute liver damage [34]. Cisplatin-induced tract of L. lanceolata leaf may have protective effect
tissue toxicity may generate reactive oxygen species against cisplatin-induced hepatorenal injuries and dyslip-
(ROS), leading to induction of hepatocyte apoptosis, the idemia in rats, which may be attributed to the phyto-
activation of inflammatory cells and their infiltration of chemicals present in it. This gives credence to the use of
tissues, as well as tissue degenerative changes [35]. this plant extract in traditional medicine for the treat-
There are two major molecular pathways of caspase- ment of diseases in humans. Further studies may be con-
dependent apoptosis which are intrinsic and extrinsic ducted in order to isolate and characterize the bioactive
[36], however the authors propose that the mechanism of compounds responsible for the pharmacological activ-
cell death in this study may be by the intrinsic pathway. ities of this plant extract, and to investigate its mechan-
This pathway is activated in response to DNA damage, oxi- ism of protection against drug-induced tissue damage in
dative stress and other stress-inducing conditions. The animals.
multiple forms of stress converge on the mitochondria and
Abbreviations
cause mitochondrial outer membrane permeabilization AC: Atherogenic coefficient; AIP: Atherogenic index of plasma; B. wt.: Body
(MOMP), which leads to dissipation of the mitochondrial weight; CIS: Cisplatin; CR: Classical ratio; CRR: Cardiac risk ratio;
membrane potential and cessation of ATP production, as CVD: Cardiovascular diseases; CYTC: Cytochrome c; GFR: Glomerular filtration
rate; LLE: Lophira lanceolata leaf extract; LSD: Least significant difference; Mg/
well as release of a number of proteins that contribute to kg: Milligramme per kilogramme; MOMP: Mitochondrial outer membrane
caspase activation [36]. In other words, intracellular stress permeabilization; NSAID: Non steroidal anti-inflammatory drug; ROS: Reactive
induces the release of proteins (Bcl-2, Bak, Bax, tBid etc), oxygen species; SEM: Standard error of mean
which promote MOMP and release a number of proteins Acknowledgements
including cytochrome c (CYTC). The release of CYTC Not applicable.
from the mitochondria leads to formation of apoptosome
Authors’ contributions
and activation of caspase 9. Then caspases 8 and 9 activate AHA and RJO designed the experiment and supervised it. SPA and RJO
downstream caspases like the caspase 3 resulting in conducted the biochemical, histological and statistical analyses, and
Ogbe et al. Clinical Phytoscience (2020) 6:4 Page 10 of 11

interpreted the results. RJO wrote the first draft of this manuscript while AHA 16. Choji TPP, Ngokere AA, Ogenyi SI, Kumbish PR. Histoarchitectural evaluation
and SPA revised the manuscript for intellectual content. All authors read and of conventional versus two rapid microwave processing techniques. Br
approved the final draft of the manuscript. Biotech J. 2015;8(3):1–19.
17. Atawodi SE, Yakubu OE, Liman ML, Iliemene DU. Effect of methanolic
Authors’ information extract of Tetrapleura tetraptera (Schum and Thonn) Taub leaves on
Not applicable. hyperglycemia and indices of diabetic complications in alloxan-induced
diabetic rats. Asian Pac J Trop Biomed. 2014;4(4):272–8.
Funding 18. Sasidharan S, Aravindran S, Latha LY, Vijenthi R, Saravanan D, Amutha S. In
The research was fully funded by the authors. vitro antioxidant activity and hepatoprotective effect of Lentinula edodes
against paracetamol-induced hepatotoxicity. Molecules. 2010;15(6):4478–89.
Availability of data and materials 19. Alqasoumi SI. Evaluation of the hepatoprotective and nephroprotective
The data sets used and/or analyzed during the current study are available activities of Scrophularia hypericifolia growing in Saudi Arabia. Saudi Pharm
from the corresponding author on reasonable request. J. 2014;22:258–63.
20. Rajakrishnan R, Lekshmi R, Benil PB, Thomas J, Alfarhan AH, Rakesh V, Khalaf
Ethics approval and consent to participate S. Phytochemical evaluation of roots of Plumbago zeylanica L. and
The protocols of this study were approved by the Animal Welfare and Ethics assessment of its potential as a nephroprotective agent. Saudi J Biol Sci.
Committee of the College of Veterinary Medicine, Federal University of 2017;24:760–6.
Agriculture, Makurdi, Nigeria. 21. Konda RV, Arunachalam R, Eerike M, Ramesh RK, Radhakrishnan KA,
Raghuraman PL, et al. Nephroprotective effect of ethanolic extract of Azima
Consent for publication tetracantha root in glycerol induced acute renal failure in Wistar albino rats.
Not applicable. J Tradit Complement Med. 2016;6:347–54.
22. el SM E-S, Abdel-Raouf OM, Fawzy HM, Manie MF. Comparative study of the
Competing interests possible protective effects of cinnamic acid and cinnamaldehyde on
The authors declare that they have no competing interests. cisplatin-induced nephrotoxicity in rats. J Biochem Mol Toxicol. 2013;27(12):
508–14. https://doi.org/10.1002/jbt.2 1515.
Received: 23 February 2019 Accepted: 29 December 2019 23. Akindele JA, Iyamu AE, Dutt P, Satti KN, Adeyemi OO. Ameliorative effect of
hydroethanolic leaf extract of Byrsocarpus coccineus in alcohol- and sucrose-
induced hypertension in rats. J Tradit Complement Med. 2014;4(3):177–88.
https://doi.org/10.4103/2225-4110.129562.
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