Elsevier

Biomedicine & Pharmacotherapy

Volume 111, March 2019, Pages 305-314
Biomedicine & Pharmacotherapy

Ameliorative effect of Homalium zeylanicum against carbon tetrachloride-induced oxidative stress and liver injury in rats

https://doi.org/10.1016/j.biopha.2018.12.045Get rights and content
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Highlights

  • Homalium zeylanicum showed hepatoprotective action in CCl4-induced hepatotoxicity.

  • ORAC, and CAP-e assay demonstrated the antioxidant activities of H. zeylanicum.

  • GC–MS and LC–MS analysis reported a total of 44 no. of secondary metabolites.

  • Catechol was identified as a major bioactive principle in H. zeylanicum.

Abstract

Objective is to evaluate the ameliorative effects of Homalium zeylanicum in carbon tetrachloride-induced oxidative stress and liver injury in rats. To establish the nature of antioxidant principles in the bioactive ethyl acetate fractions of bark (HZEB) and leaf (HZEL); oxygen radical absorbance capacity (ORAC) and cell-based antioxidant protection in erythrocytes (CAP-e) assays were performed. From acute toxicity study, HZEB and HZEL at 200 and 300 mg/kg b.w., were relatively safe at their effective doses. The degree of protection was measured by using biochemical parameters such as serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), alkaline phosphatase (ALP), total bilirubin (TB) and total protein (TP) contents. Hepatic markers e.g. thiobarbituric acid reactive species (TBARS), superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) were evaluated along with histopathological observations of liver tissues. Both fractions showed significant improvement in restoring SGOT, SGPT, ALP, TB and TP level. TBARS, SOD, CAT and GSH levels were significantly altered towards normal values. Both fractions at 300 mg/kg showed remarkable improvement in liver markers as compared to silymarin. Histopathological examinations showed reduction in hepatic necrosis and appeared normal hepatocellular architecture in HZEB and HZEL treated groups. In CAP-e assay, IC50 of HZEB (54.66 mg/mL) was higher than HZEL (60.88 mg/mL) and in ORAC assay, AUC of HZEB and HZEL were 33.46, 21.29 respectively and results were comparable with trolox. GC–MS and LC–MS analysis identified a total no. of 44 compounds. Few compounds were identified as bioactive compounds e.g. catechol (7.23%), tetraacetyl-d-xylonic nitrile (3%), oleic acid (0.49%), 2,6-bis(1,1-dimethylethyl)-phenol (3.71%), 3,4,5-trimethoxy-phenol (0.31%), and conifer alcohol (7.41%). The presence of antioxidant principles in both fractions were responsible for hepatoprotective activities, however, the presence of catechol (7.23%) in the bark part imparted better activities in protecting liver than leaf of H. zeylanicum.

Abbreviations

ROS
reactive oxygen species
NAFLD
non-alcoholic fatty liver disease
NASH
non-alcoholic steatohepatitis
APG
angiosperm phylogeny group
RT
retention time
m/z
mass to charge ratio
ORAC
oxygen radical absorbance capacity
CAP-e
cell-based antioxidant protection in erythrocytes
SGOT
serum glutamate oxaloacetate transaminase
SGPT
serum glutamate pyruvate transaminase
ALP
alkaline phosphatase
TB
total bilirubin
TP
total protein
TBARS
thiobarbiturc acid reactive species
AAPH
2,2′azobis-2-amidinopropane hydrochloride
DCF-DA
dichlorofluorescein diacetate
HZEB
ethyl acetate fractions of bark
HZEL
ethyl acetate fractions of leaf
CPCSEA
committee for the purpose of control and supervision of experiments on animals
IAEC
institutional animal ethical committee
LD50
lethal dose
OECD
organisation for economic co-operation and development
AAPH
22´-azobis-2-amidinopropane hydrochloride
SOD
superoxide dismutase
CAT
catalase
GSH
reduced glutathione
IC50
inhibitory concentration
PBS
phosphate buffer saline
FI
fluorescence intensity
AUC
area under curve
MDA
malondoaldehye
ABB
albumin-bound-bilirubin
UGT1A1
UDP-glucuronosyltransferase
LFT
liver function test
PUFA
polyunsaturated fatty acids

Chemical compounds studied in this article

CID: silymarn (CID: 1548994)
22′-azobis-2-amidinopropane hydrochloride (CID: 84924)
2′7′-dichlorofluorescein diacetate (CID:77718)

Keywords

Homalium zeylanicum
ORAC
CAP-e
Antioxidant
serum marker
Hepatic marker

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