Malagobadan, Johnathan A/L
(2016)
Phytochemical profiles, anti-asthmatic and immunomodulatory effects of lignosus rhinocerus in airway inflammation model.
PhD thesis, Universiti Sains Malaysia.
Abstract
Lignosus rhinocerus (L. rhinocerus) or locally known as Tiger’s Milk mushroom, is
traditionally used in the treatment of various diseases including asthma by
indigenous communities in Malaysia. However, to date, its efficacy on asthma and its
immunomodulating potential have not been confirmed by scientific studies. Also,
there is sparse information available on its active constituents. This study
investigated volatile constituents of hot water extract (HWE), cold water extract
(CWE) and crude (CRD) L. rhinocerus using gas chromatography mass spectrometry
(GC-MS). The anti-asthmatic potential of L. rhinocerus extracts was investigated on
ovalbumin (OVA)-induced airway inflammation Sprague Dawley rats.
Immunomodulatory effects of L. rhinocerus were carried out in BALB/c mice.
Sequential extractions of L. rhinocerus using five solvents (petroleum ether, diethyl
ether, hexane, ethyl acetate and methanol) were conducted prior to GC-MS analysis.
Overall, 44 constituents were identified from CRD L. rhinocerus. Compounds from
the fatty acid group were the most predominant (68.58%) and the main constituent
was linoleic acid (49.39%). GC-MS analysis of HWE L. rhinocerus identified 18
constituents with the main compound also linoleic acid (21.35%). In contrast, CWE
only demonstrated the presence of methylcyclopentane (97.74%) and cyclohexane
(2.26%). Male Sprague Dawley rats were sensitized with two intraperitoneal
injections of 10 mg/ml OVA emulsified in 100 mg/ml aluminium hydroxide with the
presence of 50 ng/ml of Bordetella pertussis, on days 0 and 14 followed by treatment
with L. rhinocerus extracts on day 21 for 7 days. Optimization study indicated
optimized dosage for L. rhinocerus HWE was 500 mg/kg, CWE (250 mg/kg) and
CRD (250 mg/kg). Treatments with L. rhinocerus extracts significantly ameliorated
related asthmatic parameters in the induced rats. Specifically, HWE comparatively
reduced eosinophils numbers in BALF, IgE in serum, Th2 cytokines (IL-4, IL-5 and
IL-13) levels in BALF, and eosinophil infiltrations in the lungs than the CWE and
CRD. Flow cytometry analysis revealed HWE to reduce eosinophils and improved T
regulatory cells compared to CRD and CWE. A total of 21 asthma related genes were
successfully down-regulated with HWE treatment. Immunomodulatory effects of
HWE were studied in in vitro and in vivo model. Treatment with HWE L. rhinocerus
extract demonstrated improved splenocyte population mainly of CD3+CD4+ and
CD3+CD8+ when compared with untreated mice groups. This study also
demonstrated improved splenocytes proliferation and phagocytosis activity and
controlled NO production. In conclusion, L. rhinocerus has the potential to be used
as an alternative for the treatment of allergic asthma.
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