Antioxidative and lipid-reducing activities of Ficus deltoidea in palmitic acid-induced WRL68 cells / Noor Nazirahanie Abrahim
Non-alcoholic fatty liver disease (NAFLD) is a series of liver disorder caused by deposition of fat without involving alcohol. This chronic liver disease has become a major concern globally due to its high mortality and morbidity rate. This situation leads to the quest of finding natural remed...
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| Format: | Thesis |
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2021
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| Online Access: | http://studentsrepo.um.edu.my/13249/ http://studentsrepo.um.edu.my/13249/4/nazirahanie.pdf |
| Summary: | Non-alcoholic fatty liver disease (NAFLD) is a series of liver disorder caused
by deposition of fat without involving alcohol. This chronic liver disease has become
a major concern globally due to its high mortality and morbidity rate. This situation
leads to the quest of finding natural remedy to overcome this condition. Ficus
deltoidea or commonly known as mistletoe fig or Mas Cotek, is a plant that is very
well known especially in Malaysia for its blood glucose lowering effect. However,
the potential value of F. deltoidea has not been fully evaluated, and no studies have
been published on the effects of F. deltoidea on NAFLD. Therefore, the aim of this
study is to determine the potential effect of F. deltoidea leaves on the improvement
of NAFLD especially on lipid accumulation using cultured cell model. Leaves from
three varieties of F. deltoidea, colloquially termed small-(FDS), medium-(FDM),
and big-type leaf (FDB), were subjected to water extraction. The crude (C) extracts
were fractionated using water (WF) and ethyl acetate (EAF). The phenolic and
flavonoid content, antioxidant activity, and cytotoxicity of the fractions were
investigated. Due to its high phenolic content, antioxidant activity was highest in the
FDB crude water extract, especially on metal chelating, inhibition of lipid
peroxidation, LDL oxidation and HMG-CoA reductase activities. Through MTT
assay, none of the samples showed cytotoxicity towards WRL68 cell line.
Subsequently, palmitic acid (PA) was used to induce lipid accumulation (steatosis)
in WRL68 cells, and all the samples were tested for their lipid reducing activity by
using two different protocols i.e., pre-, and post-treatments. Only FDB shows lipid
reduction at the highest tested concentration (200 µg/ml) and were used for
subsequent experiment. Pre-treatment with FDB was found to restore redox status by
increasing SOD and GPx level, and less malondialdehyde (MDA) formation. The
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changes in protein expressions were studied by using proteomic approaches via 2DE
separation followed by identification of protein spots of interest using MALDI�MS/MS. Six proteins were successfully identified which are HSPB1, PSMA1,
GSTO1, PRDX1, HIST1H2BD and UCHL3. Through bioinformatics analysis, these
proteins were found to be significantly involved in specific pathways such as
oxidative stress (PRDX1 and GSTO1), protein homeostasis (HSPB1) and
degradation (UCHL3 and PSMA1). Meanwhile, phytochemical analysis of crude
water extract of FDB using ultra high-performance liquid chromatography (UHPLC),
reveals the presence of flavone (vitexin) and phenolic acid (p-coumaric acid). Based
on the data, the polyphenolic-rich extract of F. deltoidea might have the potential to
reduce lipid levels due to the presence of phenolic compounds with high antioxidant
activities. Results obtained suggest that this Malaysian herb have a probable
preventive effect on chronic liver disease especially in the early stage of NAFLD.
Thus, these findings might be helpful to various treatment plans for fatty liver in the
future. At the same time, it shows the practice of consuming this plant to be beneficial
for health.
Keyword : Ficus deltoidea, antioxidant, steatosis, oxidative stress, protein
homeostasis
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