Free Radicals Scavenging Potential of Buccholzia Coriacea Extract and Ameliorative Effect in Paracetamol-Induced Nephrotoxicity and ...
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International Journal of Research in Pharmacy and Biosciences
Volume 6, Issue 3, 2019, PP 1-11
ISSN 2394-5885 (Print) & ISSN 2394-5893 (Online)
Free Radicals Scavenging Potential of Buccholzia Coriacea
Extract and Ameliorative Effect in Paracetamol- Induced
Nephrotoxicity and Hepatotoxicity in Rats
Fakoya, A., Olusola, A. O*
Department of Biochemistry, Faculty of Sciences, Adekunle Ajasin University,
Akungba Akoko, Ondo State. Nigeria
*Corresponding Author: Olusola, A. O, Department of Biochemistry, Faculty of Sciences, Adekunle
Ajasin University, Akungba Akoko, Ondo State. Nigeria, Email: austinolusola@gmail.com
ABSTRACT
The present study sought to evaluate the free radical scavenging activities of ethanolic extract of
Buccholzia coriacea (EBCS) by measuring its ability to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH*)
radical, nitric oxide (NOˉ) radical, 2,2 azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS*), chelate
Fe2+ radicals, hydroxyl radical (OHˉ) and inhibit lipid peroxidation (LPO). Total flavonoids and phenolics
were also determined. Antioxidant activity of EBCS was also determined in the plasma of the rats fed with
the extract by assaying for antiradical activity against DPPH and NOˉ radicals in vitro. In vivo antioxidant
effect of EBCS was also evaluated in paracetamol treated rats. Fifteen rats were randomly divided into
three groups for this study. Group 1 received normal feed as control, group 2 received 14.30 mg/kg b.w. of
paracetamol by gavage, group 3 received 400 mg/kg b.w. of EBCS for 7 days plus paracetamol on the 8 th
day. Catalase (CAT) and superoxide dismutase (SOD) activity, and malondialdehyde (MDA) levels were
assayed in the kidney, liver and serum. Histopathological examinations of liver and kidney were also
carried out. The results showed that EBCS exhibited free radical scavenging ability in dose dependent
manner towards DPPH*, NOˉ, ABTS*, OHˉ and Fe 2+ radicals as well as inhibition of LPO. The results of
evaluation of the antioxidant potentials of the extracts in the plasma showed that they were associated with
free radical scavenging activity in vivo. Paracetamol treatment caused significant (pFree Radicals Scavenging Potential of Buccholzia Coriacea Extract and Ameliorative Effect in
Paracetamol- Induced Nephrotoxicity and Hepatotoxicity in Rats
capparaceae, locally known as ‘Uke’ (Ibo), Biochemical In-Vitro Assays
‘Uworo’ (Yoruba), Owi (Edo), (Ezekiel, 2009)
Dpph Radical Scavenging Assay
is traditionally macerated in water or local gin
as a cure for diabetes. The kola is also The in-vitro antioxidant activity of the sample
recommended in the treatment of migraine. The was determined according to the method
leaves and seed have been reported to have anti- described by Mensor et al. (2001). To 1ml of
helminthic activity (Karmeswararao, 2003) as plant extract, 1ml of ethanolic solution of 2, 2-
well as anti-microbial properties, (Nweze, 2006) diphenyl-1-picrylhydrazyl (DPPH) (0.3 mM)
an attempt has been made to investigate the was added. The mixture was incubated in the
blood glucose lowering activity of methanolic dark for 30 min. The assay was standardized
extract of seeds of B. coriacea on type 2 with Tannic acid. The absorbance of the yellow
diabetes model to ascertain the folkloric claims colour solution was read at 517 nm on a
of local healers. spectrophotometer using methanol as blank.
In the present study, an attempt has been made DPPH scavenged (%) = (A control – A sample)
to evaluate the in-vitro antioxidant activities, /A control x 100.
and in vivo antioxidant of ethanolic extract of Where A control = the absorbance of the
B,coriacea in paracetamol induced toxicity in ethanol, A sample = the absorbance of the
order to justify the folkoric claims of traditional reaction mixture.
healers..
Nitric Oxide Radical Scavenging Assay
MATERIALS AND METHODS The in-vitro nitric oxide scavenging activity was
Reagents and Chemicals estimated according to the method of Marcocci
et al., 1994). To 1ml sample, 1ml of sodium
The 2, 2-diphenyl-1-picrylhydraxyl (DPPH) and nitroprusside (10 mM, aqueous) and 1 ml buffer
Hydrogen peroxide, Potassium ferricyanide, (sodium phosphate buffer, 0.2 M) were added.
sodium carbonate, butanol, methanol, acetic The mixture was incubated at room temperature
acid, thiobarbituric acid, sodium dodecyl for 150 min. this is followed by the addition of
acrylamide sulphate, Iron ii tetraoxosulphate 0.1 ml Griess reagent. The absorbance of the
(vi) used were a product of Sigma- Aldrich, pink colour solution was read at 540 nm on a
USA. 1, 10-phnanthroline used were products of spectrophotometer. The reaction was
Merck, Germany. standardized with ascorbic acid. The pink
Collection of Plant Materials chromophore generated during diazotization of
nitrite ions with sulphanilamide and subsequent
The seeds of B. coriacea were gotten from Oja coupling with N-naphthyl ethylene diamine
Oba Market at Ikare, Ondo State. The plant was dihydrochloride (NED) was measured
identified in the Department of Plant Science spectrophotometrically at 540 nm. The in-vitro
and Biotechnology, Adekunle Ajasin NOˉ scavenging activity of the sample was
University, Akungba Akoko, Ondo State, calculated by using the following formula:
Nigeria.
Nitric oxide scavenging activity (%) = (A
Extraction Procedure control - A sample)/ A control x 100.
Cold extraction method was employed. 500 g of Where A control = the absorbance of the
the clean, air dried and pulverized plants reaction mixture in the absence of sample.
samples were weighed differently into
extraction jars respectively and 1400 ml of A sample = the absorbance of the reaction
analytical grade ethanol was added to the jars mixture.
containing B. coriacea. The extraction mixture ABTS* Radical Scavenging Activity Assay
was given constant agitation and left for 72
hours. The supernatant was decanted separately The in-vitro ABTS* scavenging activity of the
and concentrated using a rotary evaporator at 40 polyphenolics was determined according to the
0
C and the extract was freeze dried. The extract method of Re et al., 1999. The stock solutions
was packed inside an airtight sample bottle and were of 8mM ABTS and 3mM Potassium
kept at 4 0C inside refrigerator until required for persulphate. The working solution was then
various in-vitro and in vivo antioxidant prepared by mixing the two stock solutions in
assessments. equal quantity and allowing them to react for 12
hours at room temperature in the dark. To 1ml
2 International Journal of Research in Pharmacy and Biosciences V6 ● I3 ● 2019Free Radicals Scavenging Potential of Buccholzia Coriacea Extract and Ameliorative Effect in
Paracetamol- Induced Nephrotoxicity and Hepatotoxicity in Rats
sample (1mg/ml), 1ml of ABTS was added. The Hydroxyl radical scavenging activity (%) =
absorbance was read at 734nm on a [ Acontrol-Asamole/Acontro] x 100.
spectrophotometer. Trolox was used as standard.
Where Acontrol= Absorbance of the control.
The in-vitro ABTS* radical scavenging activity
of the sample was calculated by using the Asample = Absorbance of plant extract sample.
following formula: Determination of Total Phenolic Content
ABTS* scavenging activity (%) = (A control -
Folin Ciocalteu reagent was used for analysis of
A sample)/ A control x 100.
total phenolics content (TPC) according to Chun
Where A control = the absorbance of the et al. (2003). In a 10 ml volumetric flask, a 0.2
reaction mixture in the absence of sample. ml aliquot of the extract in ethanol (1.0 mg/ml)
A sample = the absorbance of the reaction was mixed with 0.4 ml of Folin-Ciocalteu
mixture. reagent. The solution was allowed to stand at
25oC for 5-8 min before adding 0.2 ml of 4.0 ml
Inhibition of Lipid Peroxidation Assay of sodium carbonate solution 7.0 % and made to
In-vitro inhibition of lipid peroxidation was 10 .0 ml with distilled water. The mixture was
estimated according to the method of Ruberto allowed to stand for 2 h before its absorbance
and Baratta, 2000. In this assay, egg yolk was measured at 725 nm. Tannic acid was used
homogenate served as lipid rich medium, and as standard for the calibration curve. TPC was
FeS04 acts as initiator of lipid peroxidation. expressed as mg tannic acid equivalents (TAE)
Briefly, 40 μl of plant extract was mixed with per gram of sample (mg/g).
0.25 ml 10% egg yolk. This was followed by the Determination of Total Flavonoid Content
addition of 10 ul FeS04 (0.07 M, aqueous). The
The total flavonoid contents were measured by a
mixture was incubated at room temperature for
colorimetric assay (Zhishen et al. 1999; Zou et
30 min. This was followed by the addition of
al., 2004). A-100.0 μl aliquot of extract in
0.75 ml of glacial acetic acid (5%,v/v aqueous)
ethanol was added to a 10 ml volumetric flask
and 0.75 ml of thiobarbituric acid 0.6% in 0.2 M
containing 4 ml of distilled water. At zero time,
NaOH .The mixture was incubated in a boiling
0.3 ml 5% sodium nitrite was added to the flask.
water bath (90oC) for 20 min, cooled and
After 5 min, 0.3 ml of 10% aluminium chloride
centrifuged at 3000 rpm. One milliliter (1 ml) of
was added. At 6 min, 2 ml of 1 M sodium
the pink colour supernatant was read at 532 nm
hydroxide was added to the mixture.
on a spectrophotometer. The assay was
Immediately, the mixture was diluted to volume
standardized using quercetin.
with the addition of 2.4 ml distilled water and
In-vitro inhibition of lipid peroxidation = (A thoroughly mixed. Absorbance of the mixture,
contol – A sample)/A control X 100. Where A pink in color, was determined at 510 nm versus
control = absorbance of the control, A sample is a blank containing all reagents except sample of
the absorbance of the sample. extract. Rutin was used as standard for the
calibration curve. Total flavonoid content of the
Determination of Hydroxyl Radical Scavenging
extract was expressed as mg rutin equivalents
Activity
(RE) per gram of sample (mg/g).
The hydroxyl radical scavenging activity of the
Determination of Fe2+ Chelation Activity
extract was determined spectrophotometrically
according to the method describe by Wyu et al., The metal chelation activity of ethanolic extract
(2004). 0.09 ml of 1,10 phenanthroline was of Buccholzia coriacea was carried out
added to 1.5 ml of varying concentrations of according to the method described by Mnnoti
manitol standard (25-400 mg/ml) and the extract and Aust, 1958, modified by Pentel et al., 2005.
(25-400 mg/ml) in their respective test tubes The assay was standardised using EDTA. 900 µl
arranged in triplicate. This was followed by 0.06 of 500 µM FeSO4 was added into 150 µl of
ml of FeSO4, 0.015 ml of H2O2 and 2.4 ml of 0.2 varying concentrations (25 mg/ml – 400 mg/ml)
M phosphate buffer. Allowed to incubate for 5 of the extract. 78 µl of 0.25% w/v 1, 10-
minutes, absorbance readings were taken at 540 phenanthroline was added. Incubate at room
nm against the methanol blank. The percentage temperature for 5 minutes. EDTA was used as
hydroxyl radical scavenging effect of plant standard. The absorbance was read against the
extract was calculated as follows. blank at 510 nm.
International Journal of Research in Pharmacy and Biosciences V6 ● I3 ● 2019 3Free Radicals Scavenging Potential of Buccholzia Coriacea Extract and Ameliorative Effect in
Paracetamol- Induced Nephrotoxicity and Hepatotoxicity in Rats
The metal chelation activity (%) = Ao – As/Ao 6M H2SO4 was added to the sample, blank and
x 100. standard tubes while 0.05 M phosphate buffer
Where, Ao = absorbance of control; As = pH 7.4 (for the standard tube) was added and
absorbance of the sample. thoroughly mixed by inversion and thereafter
7.0 ml 0.01 M KMNO4 was added to all the
Estimation of Malondialdehyde (MDA) Status
tubes. Finally, the content of each tube was
The assay method of Buege and Aust, 1998 was thoroughly mixed by inversion and the
adopted. absorbance read at 480nm after 30-60 seconds
Principle against distilled water as blank. Catalase activity
was calculated.
MDA which is formed from the breakdown of
polyunsaturated fatty acid serves as a Estimation of Superoxide Dismutase (SOD)
convenient index for the determination of the Activity
extent of lipid peroxidation (LPO). MDA reacts This was determined according to the method of
with thiobarbituric acid to give a red product Misra and Fridovich (1972).
absorbing at 535nm.
Principle
Procedure
Adrenaline auto-oxidizes rapidly in aqueous
To 1 ml sample, 2 ml of TBA-TCA-HCL solution to aderenochrome, whose concentration
Reagent (ml) was added, and 3 ml of the reagent can be determined at 420nm.The auto-oxidation
was added to the blank test tube. The contents of of adrenaline depends on the presence of
each test-tube was heated for 15 minutes in
superoxide anions. Superoxide Dismutase
boiling water. After cooling, each tube was
catalyzes the breakdown of superoxide anions
centrifuged at 4000rpm for 10 minutes to
thus inhibiting the auto-oxidation of adrenaline.
remove flocculent precipitates. Absorbance of
The degree of inhibition is thus a reflection of
each supernatant was read at 535nm against the
the activity of SOD and is determined as one
blank.
unit of the enzyme activity.
Estimation of Catalase (CAT) Activity
Procedure
This was determined according to the method of
Cohen et al., 1970. 3 ml of the sample tubes contained 0.2 ml
sample, 2.5 ml of 0.05 M carbonate buffer
Principle pH10.2 and 0.3 ml of 0.03 mM adrenaline. The
Catalase catalyses the breakdown of hydrogen blank tubes contained 3.0 ml of distilled water
peroxide introduced in the presence of while the standard tubes contained 0.2 ml
phosphate buffer to water and oxygen. distilled water, 2.5 ml of 0.05M carbonate
buffer pH10.2 and 0.3 ml of 0.03 mM
2H2O2 2H2O + O2 adrenaline. The content of each tube was mixed,
The reaction is thereafter quenched (stopped) by and absorbance read at 420 nm. SOD activity
the addition of H2SO4. The amount of H2O2 was calculated.
remaining in the reaction mixture after few
Histological Test Preparations
minutes of catalase action will be determined by
titration with potassium permanganate The liver and kidney were carefully removed
(KMNO4), a very strong oxidizing reagent. The and piece of the tissues were cut and kept in
amount of substrate remaining in the mixture is 10% formalin in carefully labeled universal
inversely proportional to the activity of the containers for preservation prior to processing.
enzyme. The slides were then stained using the
Procedure Haematoxylin-Eosin staining technique through
increasing grade of alcohol and xylene before
To 0.5 ml of the sample, 5 ml of 30 mM H2O2 being mounted with a cover strip dried before
was added while the blank tube contains 0.5 ml viewing microscopically for possible malignant
distilled water. The contents of the tubes were
changes.
mixed and left standing for three minutes,1.0 ml
4 International Journal of Research in Pharmacy and Biosciences V6 ● I3 ● 2019Free Radicals Scavenging Potential of Buccholzia Coriacea Extract and Ameliorative Effect in
Paracetamol- Induced Nephrotoxicity and Hepatotoxicity in Rats
RESULTS
Table1. Antioxidant constituents of B. coriacea
Antioxidant Values
Total phenol 18.0 mg/g of GAE
Total flavonoid 37.5 mg/g QE
Vitamin C 300 mg/100 g of Ascorbic acid
Table2. DPPH free radical scavenging effect of ethanolic extract of B. coriacea
Drug Dose (µg/ml) DPPH (measured at 518 nm) % Inhibition
Control - 0.763±0.009 -
B.coreacea 25 0.476±0.004 37.78
50 0.404±0.006 47.23
100 0.314±0.003 58.91
200 0.224±0.008 70.72
400 0.063±0.004 91.76
Tannic acid 25 0.571±0.003 25.32
50 0.488±0.015 36.25
100 0.345±0.011 54.95
200 0.120±0.009 84.27
400 0.046±0.005 93.99
Table3. Nitric oxide scavenging effect of ethanolic extract of B. coreacea.
Drug Dose (µg/ml) Nitric oxide scavenging % Inhibition
Control - 0.547±0.021
B. coriacea 25 0.087±0.013 84.14
50 0.079±0.021 85.71
100 0.072±0.015 86.98
200 0.083±0.008 84.98
400 0.066±0.005 87.93
Vitamin C 25 0.302±0.002 44.79
50 0.250±0.001 54.59
100 0.106±0.005 80.80
200 0.077±0.004 85.95
400 0.035±0.005 93.58
Table4. ABTSˉ free radical scavenging effect of ethanolic extract of B. coriacea
Drug Dose (µg/ml) ABTSˉ (measured at 734nm) % Inhibition
Control - 0.403±0.002 -
B.coriacea 25 0.395±0.003 02.07
50 0.383±0.008 04.88
100 0.377±0.004 06.37
200 0.351±0.007 12.16
400 0.311±0.006 22.91
Trolox 25 0.224±0.004 44.49
50 0.086±0.004 78.66
100 0.034±0.004 91.65
200 0.008±0.001 98.09
400 0.001±0.000 99.75
Table5. Inhibition of lipid peroxide (LPOˉ) formation by the ethanolic extract of B. coriacea
Drug Dose (µg/ml) Lipid peroxide formation % Inhibition
Control - 0.577±0.004 -
B. coriacea 25 0.253±0.004 56.15
50 0.226±0.008 60.83
100 0.154±0.005 73.37
200 0.110±0.001 80.94
400 0.082±0.002 85.73
Quercetin 25 0.357±0.006 38.19
50 0.315±0.003 45.35
100 0.254±0.015 55.92
200 0.142±0.003 75.33
400 0.101±0.001 82.50
International Journal of Research in Pharmacy and Biosciences V6 ● I3 ● 2019 5Free Radicals Scavenging Potential of Buccholzia Coriacea Extract and Ameliorative Effect in
Paracetamol- Induced Nephrotoxicity and Hepatotoxicity in Rats
Table6. Hydroxyl (OHˉ) free radical scavenging effect of ethanolic extract of B. coriacea
Drug Dose OHˉ % Inhibition
(µg/ml) (measured at 510nm)
Control - 1.024±0.020 -
B.coriacea 25 0.038±0.006 96.26
50 0.038±0.003 96.26
100 0.043±0.0005 95.77
200 0.041±0.010 95.99
400 0.049±0.004 95.25
Mannitol 25 0.894±0.004 12.66
50 0.788±0.003 23.01
100 0.605±0.005 40.89
200 0.430±0.002 57.98
400 0.306±0.004 70.38
Table7. Metal chelaing effect of ethanolic extract of B. coriacea
Drug Dose ABTSˉ % Inhibition
(µg/ml) (measured at 734nm)
Control - 0.784±0.012 -
B.coriacea 25 0.739±0.008 09.90
50 0.722±0.004 11.86
100 0.476±0.004 41.88
200 0.475±0.002 41.96
400 0.417±0.007 49.03
EDTA 25 0.629±0.006 23.21
50 0.585±0.004 28.57
100 0.420±0.009 48.75
200 0.368±0.003 55.09
400 0.204±0.004 75.07
Table8. DPPH free radical scavenging activity of plasma from rats treated with ethanolic extracts of B.
coriacea
Drug Dose DPPH % Inhibition
(mg/ml) (measured at 518nm)
Control - 1.649±0.064a -
Normal - 0.836±0.070b 49.30
B.coriacea 400 0.349±0.004c 78.84
Values with different superscript are significant different at p≤0.05, Mean ± SD; n=3
Table9. Nitric oxide free radical scavenging activity in the plasma of rats treated with ethanolic extract of B.
coriacea.
Drug Dose (mg/ml) Nitric oxide scavenging activity % Inhibition
Control - 1.660±0.038a -
Normal - 0.549±0.049b 66.93
B.coriacea 400 0.317±0.003c 80.90
Values with different superscript are significant different at p≤0.05, Mean ± SD; n=3
Table10. Effect of ethanolic extract of Buccholzia coriacea on changes in SOD activity induced by
Paracetamol.
SOD/Treatments SOD in Liver SOD in Kidney SOD in Serum
(Units/mg tissue) (Units/mg tissue) (Units/ml) (n=5)x103
Control 28.802±2.919bc 30.400±0.894c 05.000±0.000a
Paracetamol only 22.668±3.653a 22.668±3.653a 05.000±0.000a
B.C + 31.656±1.775c 26.670±0.000b 05.000±0.000a
Paracetamol
Values with different superscript are significant different at p≤0.05, Mean±SD; n=5
Key: B.C = Buccholzia coriacea
6 International Journal of Research in Pharmacy and Biosciences V6 ● I3 ● 2019Free Radicals Scavenging Potential of Buccholzia Coriacea Extract and Ameliorative Effect in
Paracetamol- Induced Nephrotoxicity and Hepatotoxicity in Rats
Table11. Effect of ethanolic extract of Buccholzia coriacea on changes in MDA status induced by Paracetamol
MDA in liver (Units/mg MDA in kidney (Units/mg MDA in serum (Units/ml
Treatments tissue) Mean±SD (n)x10-5 tissue) Mean±SD (n)x10-5 serum) Mean±SD (n)x10-5
Control 2.400±1.226ab 1.755±0.219a 0.481±0.258a
Paracetamol only 3.650±0.435b 5.183±0.640c 0.853±0.447b
B.C + Paracetamol 2.885±1.933ab 1.794±0.657a 0.506±0.185a
Values with different upper case are significant different at p≤0.05, Mean±SD; n=5.
Key: B.C = Buccholzia coriacea
Table12. Effect of ethanolic extracts of Buccholzia coriacea, Psidium guajava and Morinda lucida on changes
in CAT Activity induced by Paracetamol.
CAT Treatments CAT in Liver (Units in mg tissue) CAT in kidney (Units in mg tissue) (n)x
(n)x 10-1 K/minute 10-1 K/minute
Control 0.433±0.012b 0.442±0.013 ab
Paracetamol only 0.382±0.033a 0.465±0.015b
B.C + Paracetamol 0.418±0.015b 0.452±0.023b
Values with different upper case are significant different at p≤0.05, Mean±SD; n=5
Key: B.C = Buccholzia coriacea,
Effect of Ethanolic Extracts of Buccholzia Coriacea on Liver and Kidney Ultrastucture in
Paracetamol Induced Oxidative Stress
Histological Readings Gotten from the Rat’s Liver Examinations
Results on Liver
Treatments Observations
Untreated control No lesions seen
Paracetamol only There was focal hepatocyte necrosis with heavy infiltrates of polymorphs.
Paracetamol + B. coriacea There was reduction in the area of necrosis and inflammation to a
considerable degree.
Histological Readings Gotten from the Rat’s Kidney Examinations
Results on Kidney
Treatments Observations
Untreated control No lesions seen
Paracetamol only There was an induction of in the patchy areas of acute tubular necrosis.
Paracetamol + B. coriacea Reduction in the tubular necrosis to focal area.
DISCUSSION al.,1997). The toxicity of NO increases greatly
B. coriacea has considerable amounts of phenols when it reacts with superoxide radical, forming
and flavonoids as seen in the table 1 above. the highly reactive peroxynitrite anion (ONOO-)
(Hue and Padjama, 1993). The nitric oxide
Nitric oxide is an essential bioregulatory
generated by sodium nitroprusside reacts with
molecule required for some physiological
oxygen to form nitrite. The extracts inhibit
processes like neural signal transmissions,
nitrite formation by directly competing with
immune response, control vasodilation and
oxygen in the reaction with nitric oxide. Table 1
control of blood pressure (Palmer et al., 1987;
illustrates the % inhibition of nitric oxide
Rees et al., 1989; Bredt and Synder, 1990; Gold
generation by the extract: B. coriacea and
et al., 1990). Nitric oxide has an important role
ascorbic acid in concentrations 25, 50, 100, 200,
in various inflammatory processes. Excessive
and 400 µg/ml which significantly scavenged in
production of this radical is directly toxic to
dose dependent manner with an IC50 of 21
tissues and contribute to the vascular collapse
µg/ml by B. coriace and 45.8 µg/ml by ascorbic
associated with septic shock, whereas chronic
acid. B. coriacea exhibited the better inhibition
expression of nitric oxide radical is associated
of nitric oxide formation in vitro when
with various carcinomas and inflammatory
compared with the ascorbic acid standard
conditions including juvenile diabetis, multiple
pFree Radicals Scavenging Potential of Buccholzia Coriacea Extract and Ameliorative Effect in
Paracetamol- Induced Nephrotoxicity and Hepatotoxicity in Rats
The ABTS- (2, 2’-azinobis-3-ethyl- acid in joint is an important feature of disease.
benzothiozoline-6-sulphonic Acid) radical This plant was able to scavenge OH- radicals at
reactions involve electrons transfer and the lower doses better than the standard mannitol.
process take place at faster rate when compared
Ferrozine can quantitatively form complexes
to DPPH- radicals. Re et al., 1999 reported that
with Fe2+. The complex formation can be
the decolouration of the ABTS-+ radicals also
disrupted by the presence of other complexing
reflects the capacity of an antioxidant species to
agents which cause a decrease in the red colour
donate electrons or hydrogen atoms to inactivate
this radical species. In the ABTS- radical cation intensity of complexes. Substances or samples
scavenging activity, the extracts showed that can reduce its colour intensity can be
concentration dependent scavenging activity. considered as antioxidant through the
The percentage inhibition was observed to be mechanism of inhibition of heavy metal. It was
concentration dependent. The IC50 of the extract reported that chelating agents, which form σ-
is >400 µg/ml B. coriacea and 28.10 µg/ml bonds with a metal, are effective as secondary
Quercetin standard. Buccholzia coriacea antioxidants because they reduce the redox
exhibited a weak inhibition of ABTS* radical potential, thereby stabilizing the oxidized form
when compared with the standard quercetin. of the metal ion (Kumaran and Karunakaran,
2006). Our results showed that the extract is not
1,1-diphenyl-2-picrylhydrazyl (DPPH) has been as potent as the standard EDTA, hence low or
used extensively as a free radical to evaluate weak metal chelating activity.
reducing substances. In this model it was
observed that the tested extracts showed It is known that paracetamol (PCM) induces
scavenging ability of DPPH* radicals in dose liver injury through the action of its toxic
dependent manner with IC50 values of 52.80 metabolite, N-acetyl-benzoquinoneimine,
µg/ml B. coriacea and 90.99 µg/ml Tannic acid produced by the action of Cytochrome P-450.
as standard. The extract showed better This metabolite reacts with reduced glutathione
scavenging activity when compared with the (GSH) to yield non-toxic 3-GS-ylparacetamol.
standard pFree Radicals Scavenging Potential of Buccholzia Coriacea Extract and Ameliorative Effect in
Paracetamol- Induced Nephrotoxicity and Hepatotoxicity in Rats
The decrease values of MDA formed in group enzymes activities. Similar results on PCM
treated with extract plus paracetamol when induced collapse of the antioxidant defense had
compared with paracetamol treated only group earlier been reported (Uma et. al., 2010; Sabina
showed that the extract was able to offer et. al., 2013; Dash et. al., 2007; Sowemimo et.
protection against cytotoxicity presented by the al., 2007). This effect of PCM was well
paracetamol. In the serum, kidney and liver tolerated by experimental animals receiving
tissues of rats treated with paracetamol only, extract of M. lucida hence, preventing the
there were profound injuries to the tissues as collapse of the antioxidant enzymes SOD and
seen from the histopathology results. However, CAT (Fogha et. al., 2015). The observed
the damage was ameliorated in the group treated increase of SOD activity suggests that the
with extract plus paracetamol. This implied that aqueous extract of M. lucida stem bark have an
the extract was able to show protective effect efficient protective mechanism in response to
possibly by the process of antioxidant due to the oxidative stress and may be associated with
presence of phenolic compounds in them. decreased oxidative stress and free radical-
mediated tissue damage (Fogha et. al., 2015).
Catalase is a phase II enzyme involved in the
conversion of hydrogen peroxide to water and Super oxide dismutase (SOD) is associated with
oxygen. In this study, the level of catalase increase in ROS which arises from adverse
activity was low in the liver of rats treated with metabolic activities such as lipid peroxidation.
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Citation: Fakoya, A., Olusola, A. O " Free Radicals Scavenging Potential of Buccholzia Coriacea Extract
and Ameliorative Effect in Paracetamol- Induced Nephrotoxicity and Hepatotoxicity in Rats", International
Journal of Research in Pharmacy and Biosciences, vol. 6, no. 3, pp. 1-11, 2019.
Copyright: © 2019 Olusola, A. O. This is an open-access article distributed under the terms of the
Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in
any medium, provided the original author and source are credited.
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