Metabolomics analysis of the anti-inflammatory activity of Scurrula ferruginea (Jack) danser parasitizing on three different host plants, and insights into possible mechanism
Scurrula ferruginea (Jack) Danser of Loranthaceae family is a hemi-parasitic shrub that grows on a dicotyledonous tree. Despite its traditional use for a long time in some disorders, there is very limited research on its bioactivities and phytochemistry, as well as, there is also no research conc...
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| Format: | Thesis |
| Language: | English |
| Published: |
2020
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/99219/1/IB%202020%2017%20IR.pdf http://psasir.upm.edu.my/id/eprint/99219/ |
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| Summary: | Scurrula ferruginea (Jack) Danser of Loranthaceae family is a hemi-parasitic shrub
that grows on a dicotyledonous tree. Despite its traditional use for a long time in some
disorders, there is very limited research on its bioactivities and phytochemistry, as
well as, there is also no research concerning the influence of the host plants on the
chemistry and bioactivity of S. ferruginea. The main purpose of this study was to
evaluate the anti-inflammatory activity of S. ferruginea and the mechanism of action,
as well as determine the relationship between the metabolites of S. ferruginea with its
anti-inflammatory potency. The anti-inflammatory activity was assessed via inhibition
of nitric oxide (NO) production in lipopolysaccharide (LPS) and interferon-γ (IFN-γ)
induced RAW 264.7 macrophage cells. The air dried and freeze dried S. ferruginea
parasitizing on Vitex negundo were investigated firstly. The results showed freeze
drying was the most appropriate drying method for the anti-inflammatory activity.
Subsequently, the anti-inflammatory ability was evaluated on the freeze dried leaves
and stems of S. ferruginea parasitizing on three different hosts. The results showed
that S. ferruginea stems parasitizing on Tecoma stans and Vitex negundo exhibited
higher anti-inflammatory activity compared to the corresponding samples of
harvesting from Micromelum minutum, and to the corresponding S. ferruginea leaf
samples. Their IC50 values of the two S. ferruginea stems were 114.47 ± 2.96 and
118.87 ± 2.31μg/mL, respectively. Then, the anti-inflammatory mechanism was
deciphered through messenger Ribonucleic acid (mRNA) and protein expression of
the inflammatory cytokines, including inducible nitric oxide synthase (iNOS),
cyclooxygenase-2 (COX-2), interleukin (IL), and tumor necrosis factor (TNF). The
techniques of reverse transcriptase and real time quantitative polymerase chain
reactions (RT-PCR and qPCR) were employed. The stem of S. ferruginea hosted on
Tecoma stans exerts anti-inflammatory capability attributed to the suppression of
iNOS and IL-1β mRNA expression, as well as the protein production inhibition of IL1β, IL-6, IL-10, and TNF-α besides NO secretion inhibition. Afterwards, the
metabolite variation was examined using proton nuclear magnetic resonance (1H
NMR) and liquid chromatography mass spectrometry (LC-MS) combined with
multivariate data analysis (MVDA). The metabolomics approach was employed in
evaluating the metabolite variation and its relationship with the anti-inflammatory
activity. Principal component analysis (PCA) indicated clear discriminations among
the different plant parts and host plants based on the identified metabolites. Quercitrin,
4”-O-acetylquercitrin, catechin, gallic acid and chlorogenic acid, as well as alanine,
valine, leucine, isoleucine, malic acid, succinic acid, citric acid, and acetic acid were
found higher in the leaf extracts. While threonine, histidine, fumaric acid and choline
were found present mainly in the stems. The stem of S. ferruginea-T. stans also
possessed higher quercitrin, 4”-O-acetylquercitrin, catechin, valine, leucine,
isoleucine, fumaric acid, gallic acid, chlorogenic acid, and sucrose than the stems of
S. ferruginea-V. negundo and S. ferruginea-M. minutum. A correlation was observed
from partial least square regression (PLS) model that the anti-inflammatory bioactivity
might be associated with the presence of choline, isoleucine, catechin, leucine, and
chlorogenic acid, as well as some unidentified metabolites such as ions mass m/z at
369.1157, 401.0851, 431.1327, and 473.1794. Lastly, the bioactive-activity guided
fractionation by liquid-liquid extraction on freeze-dried stem of S. ferruginea hosted
on T. stans was carried out. The results showed that ethyl acetate fraction displayed
the highest NO inhibition with 84.8 ± 1.45% at 200 μg/mL. Chloroform fraction
displayed higher activity than n-butanol and ethyl acetate fractions at 50 μg/mL, but
CHCl3 fraction showed more cytotoxicity towards the RAW 264.7 cells. The catechin
was elucidated in the ethyl acetate fraction. This is the first report on the antiinflammatory
activity of S. ferruginea parasitizing on different host plants, as well as
the extensive study concerning its plant metabolites and their correlations with antiinflammatory
property using metabolomics. The results of this study lay the
foundation for in-depth study of S. ferruginea, and further understanding on the effect
of host plants on the bioactivity of the hemiparasitic plant. |
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