Taiwanofungus Camphoratus

Antrodia camphorata, niu-chang-chih, is a unique species of fungus found
in Taiwan . Its fruit body has been used as Chinese herb in treatment of
various diseases.Recent in vivo studies have revealed that crude extracts of
Antrodia camphorata can improve liver function, increase immunity and inhibit
the growth of carcinoma.The objective of this research is to identify
bioactive compounds from Antrodia camphoratas mycelia with pharmaceutical
effect on human liver cancer cells.We used high throughput method to screen
and search for bioactive substances from the crude extract by employing human
liver cancer cell based assay, instead of slow and expensive animal testing.
At first, we were pleased to find that, when treating Hep 3B (a human liver
cancer cell) with crude extracts of mycelia and fruit body of Antrodia
camphorata for 48 hr, the IC50 values of 44.25 and 12.22 μg/mL were obtained,
respectively.The crude extract of Antrodia camphorata was further
fractionated (two times) on a silica gel column.These separated fractions (
twelve fractions in total) were individually treated with Hep 3B cells.We
found that the fraction 6 (F6) has salient bioactivity with a significant IC50
value of 6.4614 μg/mL.The active ingredient in F6 was purified by both HPLC
and TLC and structurally identified by FAB MS, NMR and IR.The compound
identified was 1-hydroxy-3-isobutyl-4-[4-(3-methyl-but-2-enyloxy)-phenyl]-
pyrrole-2,5-dione (formula: C19H23NO4 molecular weight: 329.16).In the
treatment of both Hep G2 and Hep 3B cells for 48 h, this compound gave the
IC50 values of 9.903 and 6.388μg/mL, respectively.
Antrodia camphorata, an unique species in Taiwan growing on the wood of
Cinnamomum kanehirai, is a kind of newly-developed health food. The wild type
of Antrodia camphorata is on the verge of extinction and is hardly to acquire
so the price is scorchingly high as well. Nowadays it has already been listed
as protective species. Antrodia camphorata has been developed in the culture
through deep fermentation. And the majority of scientists study liquified deep
fermentable mycelia of Antrodia camphorata as their research materials. On the
contrary solid culture is a newly-developed technique. Antrodia camphorata
cultivates by this method approximates to wild-type ones. We can expect it is
similar to natural Antrodia camphorata in therapeutic effects. Thus, we used
solid-cultured Antrodia camphorata as the research material in this study.
For the present the constituent and bioactivity of Antrodia camphorata is
still in the analysis. And now researches in polysaccharides and triterpenoids
are mainstream of in effective constituent. There have been some medical
references pointing out polysaccharides have effected on strengthening human
immunity and resisting cancers. This study explored the methanol extracts of
Antrodia camphorata effected on anti-mutation and anti-aggregation.
This study adopted Ames test, a kind of bioassay, to explore the effects
against oxidative mutation. We used Salmonella typhimurium TA102 as the test
strain. Besides, we used hydrogen peroxide, hydroxyl free radicals and cumene
hydroperoxide as mutagens to explore whether triterpenoids extracted by
methanol have expectant effects on free radical scavenging. The result of this
study is as follows The suppression ratios of the treatment of Antrodia
camphorata methanol extracts at 15 mg/ml and 10 mg/ml were statistically
significant with control. Our study found out the IC50 were 10 mg/ml, 14 mg/
ml, 6mg/ml separately while we treated hydrogen peroxide, hydroxyl free
radicals and cumene hydroperoxide as mutagens.
In platelet aggregation assay, when the extract concentration of Antrodia
camphorata were 7.5 μg/ml, 11.5 μg/ml, 15 μg/ml, they can inhibit
agglutination induced by collagen (1μg/ml). The inhibitive percentages were
59.13±3.82%, 64.27±7.84%, 96.22±3.44% separtatately. The effective
constituent of Antrodia camphorata can inhibit intracellular Ca2+ mobilization
which was induced by collagen, therefore it reduced platelet agglutination and
the inhibition curves were rather dose-responsive.