Apoptosis-Inducing Effects of Sterols from the Dried Powder of Cultured Mycelium of Cordyceps sinensis

Abstract

NotesCordyceps (Chinese name ), mainly distributed in Qinghai, Xizang, and Sichuan provinces of China, has been used as a popular and effective antiaging and tonic folk medicine and to treat various human diseases such as hepatitis, cancer, etc. Various biological activities of Cordyceps including antitumor effects were reported. [1][2][3] Recently, Cordyceps sinensis has been cultured to fulfill medicinal needs, since Cordyceps is very limited in nature. 4,5) Among the chemical constituents of cultured C. sinensis mycelium, cordycepin (6), dioxopiperazines, polysaccharides, and peptides were reported.5-7) However, sterol constituents such as 5a,6a-epoxy-5a-ergosta-7,22-dien-3b-ol (4) and ergosterol (5), which were reported to be contained in Cordyceps, 8,9) have not been isolated from the mycelium so far.We previously reported the inhibitory effects of the hotwater extract of C. sinensis mycelium on the metastasis of B16 melanoma cells in mice.10) As a continuing study, the methanolic extract and its ethyl acetate-soluble fraction from the dried powder of cultured C. sinensis mycelium showed cytotoxic effects on promyelocytic leukemia HL-60 cells. Previously, several studies on the cytotoxic and apoptosisinducing effects of Cordyceps were reported.1-3) However, the active constituents of cultured C. sinensis mycelium have not been investigated sufficiently. In this paper, the cytotoxic effects of the methanolic (MeOH) extract and several sterol constituents (1-5) from the dried powder of cultured C. sinensis mycelium on HL-60 cells are described. Results and DiscussionTo investigate the cytotoxic effects of the MeOH extract and its active constituents from the dried powder of cultured C. sinensis mycelium, the viabilities of test sample-treated HL-60 cells were evaluated in the WST-8 assay. The MeOH extract (yield: 18.9% from the dried power) exhibited significant cytotoxic effects (IC 50 valueϭ199 mg/ml). The MeOH extract was then partitioned into an ethyl acetate (EtOAc)-water (H 2 O) mixture. The aqueous phase was then extracted with 1-butanol (1-BuOH), and the EtOAc-(3.8%), 1-BuOH-(5.2%), and H 2 O-(9.9%) soluble fractions were obtained. The EtOAc-and 1-BuOH-soluble fractions showed cytotoxic effects (IC 50 values of 120 and 199 mg/ml, respectively), and the H 2 O-soluble fraction showed less potent effects (IC 50 value Ͼ200 mg/ml).Through bioassay-guided isolation, 11) sitosterol (1, 0.00025%), 5a,8a-epidioxy-22E-ergosta-6,22-dien-3b-ol (2, 0.0028%), 12) 5a,8a-epidioxy-22E-ergosta-6,9(11),22-trien3b-ol (3, 0.0045%), 12) 5a,6a-epoxy-5a-ergosta-7,22-dien3b-ol (4, 0.00012%), 9) and ergosterol (5, 0.00024%) were obtained from the EtOAc-soluble fraction by ordinary-and reversed-phase silica gel column chromatography, preparative TLC, and HPLC (Fig. 1). To best of our knowledge, this is the first report of the isolation of sterol constituents (1-5) from cultured C. sinensis mycelium, not from the insect body.Next, the cytotoxic effects of the isolated sterol constituents 1-5 on HL-60 cells were examined and comp...

Interaction of ergosterol with bovine serum albumin and human serum albumin by spectroscopic analysis

“…Recent studies showed that the ergosterol from P. umbellatus had a wide range of biological activities, such as diuretic activity [8], possessing cytotoxic activity Z. Cheng [9] and anti-inflammatory activity [10]. Besides, ergosterol was one of the most important components in fungal membranes, which had numerous biological functions, viz.…”

Pharmacokinetics of ergosterol in rats using rapid resolution liquid chromatography–atmospheric pressure chemical ionization multi-stage tandem mass spectrometry and rapid resolution liquid chromatography/tandem mass spectrometry

“…The ergosterol from Polyporus umbellatus has recently reported diuretic activity in addition to possess cytotoxic activity and anti-inflammatory activity [3][4][5]. Despite these multiple biological activities, the pharmacokinetic and biochemical aspects of this compound remain unclear.…”

Pro-apoptotic effects of Paecilomyces hepiali, a Cordyceps sinensis extract on human lung adenocarcinoma A549 cells in vitro

“…[3] CS and its derivatives are strong inhibitors of IB degradation and moderate inhibitors of IB phosphorylation. [16] All these properties together with recent advancement in fermentation, isolation and structure elucidation technologies have made research into CS derivatives seem worthwhile.…”

“…[10][11][12] Perhaps most intriguing are reports that extracts of this fungus promote apoptosis. [13][14][15][16][17][18][19] The mechanisms behind the anticancer and immune-modulating activities after treatment with CS are attributed to the production of polysaccharides, sterols, lipids, nucleosides and deoxy-nucleosides from cells. [20][21][22] CS derivatives have been produced via aseptic mycelia cultivation, the two most well-studied of which are Paecilomyces hepiali (PH; strain CS-4) and Cephalosporium sinensis.…”