Food

Chemistry
Food Chemistry 100 (2007) 16911696
www.elsevier.com/locate/foodchem

Anti-androgenic activities of the triterpenoids fraction of

Ganoderma lucidum
Jie Liu a, Kuniyoshi Shimizu a, Fumiko Konishi b, Kiyoshi Noda b, Shoichiro Kumamoto b,
Kenji Kurashiki c, Ryuichiro Kondo a,*
a
Department of Forest and Forest Products Sciences, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan
b
Chlorella Industries Co. Ltd., Research Laboratories, Fukuoka 833-0056, Japan
c
Kurume Research Park Co. Ltd., Fukuoka 839-0864, Japan

Received 20 June 2005; accepted 4 January 2006

Abstract

The ethanol extract of Ganoderma lucidum showed inhibitory activity on both isozymes (types 1 and 2) of 5a-reductase and suppres-
sion eects of ventral prostate growth induced by testosterone in castrated rat, but not induced by dihydrotestosterone. Activity-guided
fractionation and TLC analysis suggested that the active principles in vivo were triterpenoids. These results indicate that the triterpenoids
fraction of G. lucidum might be a useful ingredient in the treatment of benign prostatic hyperplasia.
 2006 Elsevier Ltd. All rights reserved.

Keywords: 5a-Reductase; Ganoderma lucidum; Anti-androgen activities; Benign prostatic hyperplasia; Triterpenoid

1. Introduction localization (Jenkins et al., 1991). Both isozymes are over-

expressed in BPH tissue (Iehle et al., 1999). Because BPH
Nowadays androgen-mediated diseases such as prostate therapy can reduce DHT levels by blocking its conversion
cancer, hirsutism, acne, androgenic alopecia, and benign from testosterone, 5a-reductase inhibitors could be useful
prostatic hyperplasia (BPH) are considered to be a serious in the treatment (Bartsch, Rittmaster, & Klocker, 2000).
problem (Bartsch, Rittmaster, & Klocker, 2002; Wasser & A number of compounds have been identied as that,
Weis, 1999). Above all, BPH is one of the most common including both a steroidal and a nonsteroidal inhibitor.
symptoms seen in older men, and 40% of men 5060 years However, it has been reported that these inhibitors may
of age and 90% of those 8090 years of age are diagnosed cause adverse eects such as gynecomastia, impairment
with BPH. The principal prostatic androgen is dihydrotes- muscle growth, and severe myopathy (Uygur, Gur, Arik,
tosterone (DHT), which is synthesized by steroid enzyme Altug, & Erol, 1998). Therefore, the emergence of thera-
5a-reductase from its substrate testosterone (Russell & peutic materials with fewer side eects, especially edible
Wilson, 1994). Since the weight of the seminal vesicles natural products, has been considered desirable if the
depends on the 5a-reduced androgens, it is important to safety of these products can be guaranteed. For thousand
maintain adequate levels of the DHT. Two isoforms of of years, mushrooms have been known to be a source of
5a-reductase have been cloned, expressed, and character- medicine. They are widely sold as nutritional supplements
ized (types 1 and 2) that display dierent tissue expression and touted as benecial for health. We have therefore
patterns, enzyme kinetic parameters, and chromosomal focused on edible and medicinal mushrooms as possible
sources of 5a-reductase inhibitory ingredients.
*
Corresponding author. Tel./fax: +81 92 642 2811. The fungi Ganoderma lucidum has been used for centu-
E-mail address: [email protected] (R. Kondo). ries in East Asia. Its fruiting body is called as Reishi in

0308-8146/$ - see front matter  2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2006.01.003
1692 J. Liu et al. / Food Chemistry 100 (2007) 16911696

Japan and Lingzhi in China. In these areas, G. lucidum inhibitory assay, the palmitic acid (9.3 mg), oleic acid
has been a popular folk or oriental medicine to cure vari- (9.8 mg), and linoleic acid (12.3 mg) were isolated and iden-
ous human diseases such as hepatitis, hypertension, hyper- tied by comparison with GCMS data of commercially
cholesterolemia, and gastric cancer (Wasser & Weis, 1999; available standard samples.
Yun, 1999). However, the precise mechanism and active
compounds of G. lucidum against these biological activities 2.4. Preparation of rat microsomes
have remained unclear.
In the continuing search for active principles from the Rat liver and prostate microsomes from female SD rat
ethanol extract of G. lucidum against the growth inhibition (7 weeks age) and male SD rat (13 weeks age) were pre-
of the ventral prostate induced by testosterone in rat, we pared, respectively, by a method previously reported by
have found a characteristic fraction containing triterpe- Shimizu et al. with some modications (Shimizu, Fukuda,
noids after separation by silica gel column chromatography Kondo, & Sakai, 2000). Two mature SD female rats were
that showed signicant activity. killed. The liver was removed, and minced tissue was then
homogenized in a 4-tissue volume medium A (0.32 M
2. Materials and methods sucrose, 1 mM dithiothreitol, and 20 mM sodium phos-
phate, pH 6.5). Also, three mature SD male rats were
2.1. Materials killed. The prostate was removed, and minced tissues were
then homogenized in 4-tissue volume medium A. The
The fruiting body of G. lucidum was obtained from Biso- homogenate was then centrifuged at 10,000g for 10 min.
ken Inc. (Fukuoka, Japan). The fruiting bodies were dried The resulting supernatant from the centrifugations was fur-
and ground to a powder before use. Unless otherwise spec- ther centrifuged twice at 105,000g for 1 h twice. The
ied, chemicals were obtained from Sigma Chemical Co. washed microsomes were suspended in 1-pellet volume
(Japan). Organic solvents were purchased from Wako Pure medium A, and the dispersion of microsomes was achieved
Chemical Industries Co. (Japan). [4-14C] Testosterone was using a syringe with 18 G, 23 G, and 26 G needles in suc-
obtained from PerkinElmer (Japan). SpragueDawley cession. The microsome suspension was divided, diluted
(SD) rat was obtained from Charles River (Japan). Ganod- with medium A, and stored at 80 C until just before use.
erol B, ganodermanontriol, ganoderic acid G and ganod-
eric acid C2 were provided by the Laboratory of 2.5. Measurement of 5a-reductase inhibitory activity
Systematic Forest and Forest Products Sciences, Depart-
ment of Forest Products, Faculty of Agriculture, Kyushu The complete reaction mixture included 1 mM dithio-
University in Japan. threitol, 20 mM phosphate buer (pH 6.5 for type 1 or
pH 5.0 for type 2), 1.9 nCi [4-14C] testosterone, 150 lM tes-
2.2. Ethanol extracts of G. lucidum tosterone, 167 lM NADPH, and the enzyme preparation
(1.54 mg of protein) in a nal volume of 0.3 ml. The con-
Dried and chipped G. lucidum (15 kg) was extracted with centration of testosterone contributed by [4-14C] testoster-
95% ethanol (126 l) at room temperature for 24 h using a one was negligible. The incubation was carried out for
blender. The extracts were ltered through ADVANTEC 10 min at 37 C, and was started by the addition of 10 ll
No. 2 lter paper, concentrated under vacuum, and then microsomes to the preheated reaction solution in a tube.
freeze-dried. The extracts (571.1 g) were stored at 20 C After 10 min, the incubation was terminated by adding
before assay. 10 ll of 3 M NaOH. To extract the metabolites, 1 ml of
diethyl ether was added, and the tubes were capped and
2.3. Fractionation of the ethanol extracts of G. lucidum by shaken. The organic phase was applied to a silica plate
silica gel column chromatography (Kieselgel 60 F254). The plate was developed in ethyl ace-
taten-hexane (7:3) at room temperature. The radioactivity
A portion of the ethanol extracts (50 g) was fractionated prole was determined with an imaging analyzer (FLA-
into three fractions (Fr. AC) by column chromatography 5000 RF, Fuji Film Co. Ltd., Tokyo, Japan). The 5a-
on silica gel (Wakogel C-200, Wako, Osaka, Japan) (2 kg, reductase activity was calculated from the percentage of
column size; 20 cm i.d. 150 cm) eluting with an n-hexane- the extent of the conversion of [4-14C] testosterone to
EtOAc step-gradient [Fr. A: n-hexane:ethyl acetate = 9:1 [4-14C] dihydrotestosterone.
(2 l), 8:2 (2 l); Fr. B: 7:3 (2 l), 6:4 (2 l), 5:5 (2 l), 4:6 (2 l),
3:7 (2 l), 2:8 (2 l); Fr. C: 1:9 (2 l), MeOH (6 l)]. [Fr. A: 2.6. Growth suppression of the rat prostate by G. lucidum
TLC, silica gel, I2 detection, EtOAc/n-hexane, 7:3, Rf
0.970.48, Fr. B: Rf 0.670.03, Fr. C: Rf 0.040] (Fig. 3). The assay for growth suppression of the rat prostate was
These procedures were repeated 11 times and combined carried out as described by Fukuta et al. (Fukuta et al.,
into each fraction [Fr. A (240 g), Fr. B (35 g), Fr. C 1999). The testes of SD rat were removed at 4 weeks of
(269 g)]. From the further separation of a part of Fr. A age under light anesthesia with pentobarbital. After 4 days,
(1.0 g) by a silica gel column guided with a 5a-reductase testosterone (100 lg/ body) or dihydrotestosterone
 J. Liu et al. / Food Chemistry 100 (2007) 16911696 1693

(300 lg/ body) was injected s.c. into the rats once daily for tionation was carried out. The ethanol extracts were
7 days. The animals were freely administered CL-2 food roughly separated into three fractions (Fr. A, B, C), which
(Oriental yeast Co., LTD). The samples suspended in were analyzed by TLC as illustrated in Fig. 2. Fr. A and Fr.
0.5% methylcellulose were orally administered at each con- B showed 5a-reductase inhibitory activity of more than
centration once daily. The utamide (10 mg/kg/day) was 90% at 200 lg/ml, while Fr. C did not show the 5a-reduc-
used as the positive control and was suspended in 0.5% tase inhibitory activity (less than 25% inhibition at 200 lg/
methylcellulose and orally administered once daily for 7 ml) (data not shown). Most of constituents of Fr. A were
days. After 7 days, the rats were fasted for 18 h and sacri- claried to be fatty acids by GCMS analysis (data not
ced by pentobarbital. The prostate was then removed, and shown). A portion of Fr. A was submitted to further sepa-
the weight was determined. ration by silica gel CC, which led to the isolation of pal-
mitic acid (IC50 = 870 lM), oleic acid (IC50 = 42.4 lM),
2.7. Statistics and linoleic acid (IC50 = 190 lM) as 5a-reductase inhibi-
tory components.
Results were expressed as means SD. Statistical signif- Some fatty acids have been reported as 5a-reductase
icance was determined by ANOVA and Bonferroni-type inhibitory materials (Liang & Liao, 1992). Also, amongst
multiple t-test. the numerous phytotherapeutic compounds aimed at treat-
ing lower urinary tract symptoms (LUTS) secondary to
3. Results and discussion BPH, the most popular and widely used is the extract of
the dried ripe fruit of Serenoa repens (also named, Sabal
In our previous screening of 19 edible and medicinal serrulata) (Lowe & Fagelman, 1999). The lipidsterolic
mushrooms, we discovered that the ethanol extract of the extract of S. repens (LSESr, Permixon), obtained by
fruiting body of G. lucidum showed the strongest 5a-reduc- hexane extraction, is essentially composed of fatty acids
tase inhibitory activity (Fujita et al., 2005). In addition, the (a mean from 15 dierent batches of 94 g/100 g of extract),
treatment of the ethanol extract prepared from G. lucidum of which free fatty acids account for a mean of 83.5 g/100 g
at 1.5 and 15 mg/kg/day signicantly inhibited the growth of extract. The mean percentage values in the free fatty acid
of the ventral prostate induced by testosterone in rat (Fuj- fractions are 35%, 30%, 11%, and 5% for oleic, lauric,
ita et al., 2005). These results led us to further investigation myristic, and linoleic acids, respectively (Niederpruem,
of the ethanol extract of G. lucidum. The inhibition of the Schweikert, & Zaenker, 1994; Weisser, Tunn, Behnke, &
5a-reductase prepared from rat liver by the ethanol extract Krieg, 1996).
of G. lucidum was concentration-dependent, as shown in In light of these previous reports indicating that S.
Fig. 1. As the concentrations of ethanol extract increased, repens has high levels of fatty acids, fatty acids such as oleic
the residual enzyme activity rapidly decreased. The inhibi- acid, linoleic acid, and palmitic acid isolated from Fr. A
tory concentration leading to 50% activity loss (IC50) was were expected to be involved in the growth suppression
estimated to be 93.6 lg/ml. It should be noted that naste- of prostate caused by the ethanol extract of G. lucidum.
ride (Liang, Cascieri, Cheung, Reynolds, & Rasmusson, However, the treatment of Fr. A and oleic acid which
1985), which is known as a potent steroidal inhibitor,
showed an IC50 of 0.73 lM in our assay system.
To clarify the active principles of the ethanol extract of
G. lucidum, 5a-reductase inhibitory activity-guided frac-

100
90
80
Inhibitory acitivity (%)

70
60
50
40
30
11 22 33 44 55 66 77
20 1.. Fr. A, 2. Fr. B, 3. Fr. C, 4.. ganoderol B,
10 5. ganodermanontriol , 6. ganoderic acid G,
7. ganoderic acid C2
0
0 50 100 150 200
g/ml Fig. 2. TLC analysis of Fr. A, B and C prepared from the ethanol extract
of G. lucidum, and the well-known representative triterpenoids (ganoderol
Fig. 1. The concentration dependence of the inhibitory eects of the B, ganodermanontriol, ganoderic acid G and ganoderic acid C2) in
ethanol extract of G. lucidum on 5a-reductase (liver microsome, pH 6.5). Ganoderma mushroom (EtOAc:n-hexane = 7:3).
1694 J. Liu et al. / Food Chemistry 100 (2007) 16911696

showed the strongest 5a-reductase inhibitory activity 5a-reductase activity derived from both isozymes prepared
among fatty acids isolated from Fr. A, did not inhibit the from prostate (Fig. 4).
growth of the ventral prostate induced by testosterone in The reason that Fr. A did not show a growth-suppres-
rat (Fig. 3). Also, as expected, Fr. C did not show inhibi- sion eect in prostate, despite its potent 5a-reductase inhib-
tory activity in vivo. Interestingly, Fr. B showed a itory activity, remains unclear. However, it may not be
growth-suppression eect as similar to that of nasteride illogical to assume that the fatty acids are not an active
which is used as a positive control. principle in vivo since fatty acids are easily metabolized
G. lucidum has been reported to produce many bioactive into inactive compounds or do not reach the target organ
oxygenated triterpenoids. Up to now, over 120 species of by oral administration. Also, it has been reported that oral
triterpenoids have been isolated from G. lucidum and the administration of some dietary fatty acids enhances ste-
genus Ganoderma (Shiao, 2003). It is likely that most of roid-metabolizing microsomal membrane-bound enzymes,
triterpenoids are present in Fr. B by the result of TLC anal- 5a-reductase, and aromatase in rat liver (Venkatraman,
ysis (Fig. 2) of typical triterpenoids isolated from G. luci- Rao, Fink, & Awad, 1996).
dum (Shiao, 2003). For example, the Rf values of Prostatic enlargement is dependent on tissue androgen,
ganoderol B, ganodermanontriol, ganoderic acid G and namely DHT, which is converted from testosterone by ste-
ganoderic acid C2 which are known to be the typical trit- roid 5a-reductase. In the present study, we investigated the
erpenoids in Ganoderma mushroom, developed in ethyl eects of G. lucidum on steroid 5a-reductase activity and on
acetaten-hexane (7:3) are 0.64, 0.30, 0.23 and 0.07, respec- the growth of prostate induced by testosterone in castrated
tively (Fig. 2). Also, it has been conrmed that these trit- rats. The Fr. B was found to inhibit both types of 5a-reduc-
erpenoids are detected in the Fr. B by qualitative HPLC tase, and this so-called dual inhibition might be advanta-
analysis, but not quantitative analysis (data not shown). geous in the therapy for BPH, since it has been shown
Therefore, it is reasonable that we dene the Fr. B as the that the dual inhibitor dutasteride is more powerful in
triterpenoids fraction. Also, it should be noted that we reducing DHT plasma concentrations than selective type
have not examined the activities of these triterpenoids on 1 or type 2 inhibitors (Graul, Silvestre, & Castaner,
5a-reductase and animal test yet, because of fewer limited 1999). In addition, the treatment of ethanol extract of G.
amount of samples. We are still trying to isolate these trit- lucidum and Fr. B signicantly inhibited the growth of
erpenoids for further experiments. the ventral prostate induced by testosterone in castrated
Two 5a-reductase isozymes have been identied in rats rats. These results suggest that the suppression eects of
and humans (Russell & Wilson, 1994). Both isozymes are prostatic growth by G. lucidum might come in part from
overexpressed in BPH tissue (Iehle et al., 1999). Coded its ability to act as a 5a-reductase inhibitory material.
by two dierent genes (Andersson & Russell, 1990), they There are commonly two ways to suppress prostate
display a maximal activity at dierent pH values (6.08.5 regrowth in animal experiments. One is inhibiting the
for type 1 and 5.05.5 for type 2); they also have dierent 5a-reductase activity. The other is to act on the androgen
biochemical characteristics (Li, Chen, Singh, & Labrie, receptors as an androgen receptor antagonist. The
1995). In rats, the type 1 isozyme predominates in tissues androgen antagonist can suppress the dihydrotestoster-
such as liver, kidney, brain, lung, and skin, but it also exists one-induced prostate regrowth. Therefore, blocking
in prostate, whereas the type 2 isozyme is more abundant in dihydrotestosterone from binding to the androgen recep-
genital tissues such as prostate. Fr. A and Fr. B inhibited tors in the prostate grand is considered to be a possible

40 90
(mg/100g body weight )

30
Prostate weight

Inhibitory activity (%)

* * 60
20

10
30

0
C+T C+T+O C+T+Fr.A C+T+Fr.B C+T+Fr.C C+T+F
0
Fig. 3. Eects of the fractions prepared from ethanol extracts of G. Ethanol extracts Fr. A Fr. B
lucidum on testosterone-induced regrowth of the castrated rat prostrate.
Each column represents the mean SD, n = 6; C: castrated rat, T: Fig. 4. The inhibitory activity of Fr. A, Fr. B and the ethanol extracts of
testosterone, O: olenic acid (1 mg/kg of body weight), Fr. A, Fr. B, Fr. C G. lucidum on type 1 and type 2 5a-reductase. Each column represents the
(1 mg/kg of body weight), F: nasteride (1 mg/kg of body weight) mean SD. Sample concentration is 200 lg/ml, n = 3. (j): liver micro-
*p < 0.01 against C + T. some (pH 6.5 for type 1), (): prostate microsome (pH 5.0 for type 2).
 J. Liu et al. / Food Chemistry 100 (2007) 16911696 1695

mechanism of action other than 5a-reductase inhibition of patients using herbal dietary supplements is not exactly
the extract of G.lucidum. To examine this possibility, the known, there is evidence of the increasing use of dietary
eects of ethanol extracts of G. lucidum on prostate growth supplements in cancer treatment (Eisenberg et al., 1998).
induced by dihydrotestosterone have been investigated. In G. lucidum is one of the herbs in the herbal mixture PC-
other words, if the prostate suppression is caused by only SPES, which has shown activity against hormone-refrac-
5a-reductase inhibition, the dihydrotestosterone-induced tory disease in two prostate cancer patients (de la Taille,
prostatic regrowth cannot be suppressed. Four days after Hayek, Burchardt, Burchardt, & Katz, 2000). Extracts of
castration, the weights of the rat prostates were markedly PC-SPES have demonstrated estrogenic eects (DiPaola
reduced, and the prostate size was recovered by s.c. et al., 1998) and decrease the growth of hormone-sensitive
injections of dihydrotestosterone (300 lg/ body) (Fig. 5). as well hormone-insensitive prostate cancer cells (de la
Ethanol extract of G. lucidum had no eect on the weight Taille et al., 1999). Our results suggest that these eects
of the prostate in the castrated rats that received dihydro- might be related to not only the anti-cancer eects of G.
testosterone, whereas utamide (Reid, Kanto, & Oh, lucidum but also anti-androgen eects. Since excessive 5a-
1999), an androgen receptor antagonist, signicantly reductase activity has been proposed to be a possible con-
reduced the prostate weights (Fig. 5). These results suggest tributing factor in prostate cancer development and pro-
that G. lucidum inhibited prostatic growth by the inhibition gression, the development and progression of prostate
of 5a-reductase activity rather than having a direct eect cancer may also be aected by diets containing inhibitors
on the androgen receptor, although the precise mechanism of 5a-reductase.
remains unclear. In the present study, we detected anti-androgenic
The fungi G. lucidum (Reishi, Mannentake, or Lingzhi) activities of the ethanol extract of G. lucidum on in vitro
has been used for centuries in East Asia to cure various 5a-reductase inhibitory activity and in vivo growth sup-
human diseases such as hepatitis, hepatopathy, hyperten- pression of the rat prostate. Our results indicate that the
sion, nephritis, bronchitis, and cancers (Wasser & Weis, active principles in vivo might be triterpenoids in Fr. B
1999; Yun, 1999). Its dried powder was especially popular rather than fatty acids in Fr. A. In the future, herbal ther-
as a cancer chemotherapy agent in the Imperial Court of apies will likely become important treatments in wide use
ancient China (Mizushina et al., 1998). Some of the triter- for many diseases (Uygur et al., 1998). The anti-androgenic
penes such as ganoderic and lucidic acids, recently isolated activity of G. lucidum is important biological activity that
from Ganoderma, have demonstrated cytotoxicity against could be useful in BPH patients. At this time, the clinical
mouse sarcoma and mouse lung carcinoma cells in vitro implications of this activity are unknown, so further
(Min, Gao, Nakamura, & Hattori, 2000). Intraperitoneal research is needed for general use to apply G. lucidum to
administration of water-soluble polysaccharides isolated the treatment of BPH.
from Ganoderma has been found to inhibit the growth of
sarcoma-180 solid tumors in mice (Sone & Misaki, 1985).
References
In addition, polysaccharides from Ganoderma also potenti-
ate production of cytokines, which subsequently suppress Andersson, S., & Russell, D. W. (1990). Structural and biochemical
proliferation of HL-60 and U937 leukemic cell lines (Wang properties of cloned and expressed human and rat steroid 5a-
et al., 1997). reductases. Proceedings of the National Academy of Sciences of the
The use of herbal therapies in alternative medicine has United States of America, 87, 36403644.
Bartsch, G., Rittmaster, R. S., & Klocker, H. (2000). Dihydrotestosterone
been increasing, and although the number of cancer
and the concept of 5a-reductase inhibition in human benign prostatic
hyperplasia. European Urology, 37, 367380.
Bartsch, G., Rittmaster, R. S., & Klocker, H. (2002). Dihydrotestosterone
100 and the concept of 5a-reductase inhibition in human benign prostatic
(mg/kg of body weight)

hyperplasia. World Journal of Urology, 19, 413425.

de la Taille, A., Hayek, O. R., Burchardt, M., Burchardt, T., & Katz, A.
Prostate weight

75 * E. (2000). Role of herbal compounds (PC-SPES) in hormone-refrac-

tory prostate cancer: two case reports. Journal of Alternative and
50 Complementary Medicine, 6, 449451.
de la Taille, A., Hayek, O. R., Buttyan, R., Bagiella, E., Burchardt, M., &
Katz, A. E. (1999). Eects of a phytotherapeutic agent, PC-SPES, on
25
prostate cancer: a preliminary investigation on human cell lines and
patients. BJU International, 84, 845850.
0 DiPaola, R. S., Zhang, H., Lambert, G. H., Meeker, R., Licitra, E., Ra,
C+DHT C+DHT+G1 C+DHT+G10 C+DHT+Fl M. M., et al. (1998). Clinical and biologic activity of an estrogenic
herbal combination (PC-SPES) in prostate cancer. New England
Fig. 5. Eects of ethanol extracts of G. lucidum on dihydrotestosterone- Journal of Medicine, 339, 785791.
induced regrowth of the castrated rat prostrate. Each column represents Eisenberg, D. M., Davis, R. B., Ettner, S. L., Appel, S., Wilkey, S.,
the mean SD, n = 6; C: castrated rat, DHT: dihydrotestosterone, G1: Van Rompay, M., et al. (1998). Trends in alternative medicine use
ethanol extracts of G. lucidum (1.0 mg/kg of bodyweight), G10: ethanol in the United States, 19901997: results of a follow-up national
extracts of G. lucidum (10 mg/kg of body weight), F: utamide (10 mg/kg survey. JAMA: Journal of the American Medical Association, 280,
of body weight) *p < 0.01 against C + T. 15691575.
1696 J. Liu et al. / Food Chemistry 100 (2007) 16911696

Fujita, R., Liu, J., Shimizu, K., Konishi, F., Noda, K., Kumamoto, S., Niederpruem, H. J., Schweikert, H. U., & Zaenker, K. S. (1994).
et al. (2005). Anti-androgenic activities of Ganoderma lucidum. Journal Testosterone 5a-reductase inhibition by free fatty acids from Sabal
of Ethnopharmacology, 102, 107112. serrulata fruits. Phytomedicine, 1, 127133.
Fukuta, Y., Fukuda, Y., Higashino, R., Yoshida, K., Ogishima, M., Reid, P., Kanto, P., & Oh, W. (1999). Antiandrogens in prostate cancer.
Tamaki, H., et al. (1999). Z-350, a novel compound with a1- Investigational New Drugs, 17, 271284.
adrenoceptor antagonistic and steroid 5a-reductase inhibitory actions: Russell, D. W., & Wilson, J. D. (1994). Steroid 5a-reductase: two
pharmacological properties in vivo. The Journal of Pharmacology and genes/two enzymes. Annual Review of Clinical Biochemistry, 63,
Experimental Therapeutics, 290, 10131018. 2561.
Graul, A., Silvestre, J., & Castaner, J. (1999). Dutasteride: steroid 5a- Shiao, M. S. (2003). Natural products of the medicinal fungus Ganoderma
reductase inhibitor treatment of BPH. Drugs of the Future, 24, lucidum: occurrence, biological activities, and pharmacological func-
246253. tions. Chemical Record, 3, 172180.
Iehle, C., Radvanyi, F., Gil Diez de Medina, S., Ouak, L. H., Gerard, H., Shimizu, K., Fukuda, M., Kondo, R., & Sakai, K. (2000). The 5a-
Chopin, D., et al. (1999). Dierences in steroid 5a-reductase iso- reductase inhibitory components from heartwood of Artocarpus
enzymes expression between normal and pathological human prostate incisus: structureactivity investigations. Planta Medica, 66,
tissue. Journal of Steroid Biochemistry and Molecular Biology, 68, 1619.
189195. Sone, Y., & Misaki, A. (1985). Structures and antitumor activities of the
Jenkins, E. P., Hsieh, C. L., Milatovich, A., Normington, K., Berman, D. polysaccharides isolated from fruiting body and the growing culture of
M., Francke, U., et al. (1991). Characterization and chromosomal mycelium of Ganoderma lucidum. Agricultural and Biological Chemis-
mapping of a human steroid 5a-reductase gene and pseudogene and try, 49, 26412653.
mapping of the mouse homologue. Genomics, 11, 11021112. Uygur, M. C., Gur, E., Arik, A. I., Altug, U., & Erol, D. (1998). Erectile
Li, X., Chen, C., Singh, S. M., & Labrie, F. (1995). The enzyme and dysfunction following treatments of benign prostatic hyperplasia: a
inhibitors of 4-ene-3-oxosteroid 5a-oxidoreductase. Steroids, 60, prospective study. Andrologia, 30, 510.
430441. Venkatraman, J. T., Rao, M., Fink, C. S., & Awad, A. B. (1996). Eect of
Liang, T., Cascieri, M. A., Cheung, A. H., Reynolds, G. F., & Rasmusson, dietary lipids on activities of hepatic steroid metabolizing enzymes (5a-
G. H. (1985). Species dierences in prostatic steroid 5a-reductases of reductase and aromatase) and composition of microsomes. Nutrition
rat, dog, and human. Endocrinology, 117, 571579. Research, 16, 17491759.
Liang, T., & Liao, S. (1992). Inhibition of steroid 5a-reductase by specic Wang, S. Y., Hsu, M. L., Hsu, H. C., Tzeng, C. H., Lee, S. S., Shiao, M.
aliphatic unsaturated fatty acids. Journal of Biochemistry, 285, S., et al. (1997). The anti-tumor eect of Ganoderma lucidum is
557562. mediated by cytokines released from activated macrophages and T
Lowe, F. C., & Fagelman, E. (1999). Phytotherapy in the treatment of lymphocytes. International Journal of Cancer, 70, 699705.
benign prostatic hyperplasia: an update. Urology, 53, 671678. Wasser, S. P., & Weis, A. L. (1999). Therapeutic eects of substances
Min, B. S., Gao, J. J., Nakamura, N., & Hattori, M. (2000). Triterpenes occurring in higher Basidiomycetes mushrooms: a modern perspective.
from the spores of Ganoderma lucidum and their cytotoxicity against Critical Review Immunology, 19, 6596.
meth-A and LLC tumor cells. Chemical Pharmaceutical Bulletin Weisser, H., Tunn, S., Behnke, B., & Krieg, M. (1996). Eects of the Sabal
(Tokyo), 48, 10261033. serrulata extract IDS 89 and its subfractions on 5a-reductase activity in
Mizushina, Y., Hanashima, L., Yamaguchi, T., Takemura, M., Sugawara, human benign prostatic hyperplasia. Prostate, 28, 300306.
F., Saneyoshi, M., et al. (1998). A mushroom fruiting body-inducing Yun, T. K. (1999). Update from Asia. Asian studies on cancer chemo-
substance inhibits activities of replicative DNA polymerases. Biochem- prevention. Annals of the New York Academy of Sciences, 889,
ical and Biophysical Research Communications, 249, 1722. 157192.