CONCLUDING REMARKS
VIII. CONCLUDING REMARKS
G. lucidum is a well-regarded Asian herbal remedy with a long and impressive range of applications. Global consumption of G. lucidum is high, and an increasingly large series of patented and commercially available products that incorporate G. lucidum as an active ingredient are available as food supple- ments. These include extracts and isolated constituents in various formula- tions and these are marketed worldwide in the form of capsules, creams, hair tonics, and syrups (6,182). The various postulated health benefits of G. lucidum are outlined in Figure 5, and a summary of the experimental studies described in this chapter, with our comments, on the putative therapeutic effects of G. lucidum is presented in Table 2.
In conclusion, studies on G. lucidum composition, cultivation, and reputed effects are still being carried out, and several new studies have been published since this paper was drafted. However, convincing evidence of direct effects of G. lucidum on human health is lacking to date, even though the mushroom is widely promoted and consumed. Observational and anecdotal reports of benefits have not as yet been substantiated by well-controlled clini- cal trials or reliable scientific data. The G. lucidum preparations used have
Lingzhi Polyphorous Fungus (Ganoderma lucidum) 209
F IGURE 5 Postulated, but as yet unvalidated, health benefits of Lingzhi (G. lucidum).
T ABLE 2 Summary of Experimental Studies a on the Putative Therapeutic Effects of the Lingzhi Mushroom (Ganoderma lucidum)
Ref. Viral infection
Effect tested
Type of study
Reported effect
Comment
Viral proliferation
Cell culture and
Inhibitory effect against
G. lucidum may have an
in vitro
some viruses,
inhibitory effect on viral
particularly herpes
proliferation, and the
simplex; synergistic or
possible synergistic
additive effects with
effect with established
antiviral drugs reported.
antiviral drugs is
Some isolated triterpenes
interesting; further in
inhibited (HIV)-1
vitro studies are
reverse transcriptase
needed.
and protease.
Viral infection Recovery from or
46,116,117 progression of
Human
Promoted recovery from
Only 3 trials published to
intervention
postherpetic neuralgia
date; these were small,
viral infection
trials
(n = 4) and reportedly
poorly designed, and
also aided recovery in a
uncontrolled studies
small (n = 4) trial on
and do not provide
hepatitis B patients. No
convincing scientific
improvement in quality
evidence for G. lucidum
Wachtel-Galor
of life or immune status
in promoting recovery
was seen in HIV
from viral infection;
patients (n = 68) given
well-planned,
a mix of Chinese herbs
controlled clinical trials
together with lingzhi.
are needed.
Bacterial infection Bacterial
119,121 et proliferation
In vitro and
In vitro inhibitory effect
No human trials to date;
animal
reported against some
experimental evidence
al.
studies
bacteria; additive or
of possible interaction
Lingzhi
synergistic effect
with antibiotics is
reported with antibiotic
interesting, as G.
drugs, but indication of
lucidum may help lower
some antagonistic
MIC values of
Polyphorous
effect also seen.
potentially cytotoxic drugs. However, antagonistic effects must be clarified, and testing for possible
Fungus
antibacterial effect against antibiotic resistant strains would
(Ganoderma Cancer
be useful.
Tumor growth
Animal studies
Administration of G.
Antitumor activity
and human
lucidum to animals (i.p
appears related to
clinical trials
or o.p) inhibited growth
polysaccharide content
of implanted/induced
of mushroom. Only two
tumors (see Table 1).
human trials reported to
lucidum
Human studies of acute
date; there were small
myeloblastic leukemia
(n = 4 and n = 5) and
and advanced
uncontrolled, involved
nasopharyngeal
advanced illness, and
carcinoma reported
G. lucidum was given in
improvement in
conjunction with
patients treated with
standard therapy;
high-dose G. lucidum
contribution of G.
extract in combination
lucidum to reported
with conventional
improvement in
therapy.
patients is not clear.
212 T ABLE 2 Continued
Effect tested
Type of study
Reported effect
Comment
Ref.
Cancer Growth and
48,50,52,86–88, activation of
Cell culture and
Incubation with G.
Direct effects on cancer
109,121,126, cancer and
cells ex vivo
lucidum caused
cells may be due to
129—135 immune cells
and in vitro
activation of cultured
triterpene-mediated
macrophages and T
inhibition of cell division
lymphocytes; increase
and inhibition of DNA
in cytokine production;
polymerase. Indirect
activated macrophages
effects appear to be
inhibited growth of
more pronounced and
cultured cancer cells,
mediated by
promoted apoptosis
polysaccharide-induced
and differentiation;
effects on immune
effects on cancer cells
cells.
not seen with untreated macrophages or with G. lucidum alone; G. lucidum induced phase
II enzymes in cultured cells; terpenes from G.
Wachtel-Galor
lucidum reported to inhibit DNA polymerase in vitro.
Immune status Immunomodulation
Animal studies
Enhanced lymphocyte
Opposing effects—
and cells ex
proliferation, antibody
activation and
vivo
production, and
suppression—seen with
137–140 et
recovery of the immune
different constituents.
al.
system in irradiated
Studies often poorly
Lingzhi
mice reported. An
described and appear
isolated protein (LZ-8)
uncontrolled. Further
and methanolic extracts
study is needed.
of G. lucidum showed
Polyphorous
immunosuppressive effects, such as delaying rejection time of allografts in mice.
CVD Blood pressure and
7,11,79, cholesterol
In vitro and
Inhibition of cholesterol
Some evidence that G.
142–145 Fungus synthesis
animal
synthesis and ACE
lucidum has potential
studies;
demonstrated in vitro;
as a hypotensive and
human
hypotensive effect seen
hypocholesterolemic
intervention
in some animal studies;
agent; effects on blood
(Ganoderma
trial
hypotensive and small
pressure may be due to
cholesterol-lowering
ACE inhibition; effects
effect reported in
on cholesterol may be
human intervention trial
mediated by a
of 40 hypertensive
combination of
subjects treated with G.
inhibition of cholesterol
lucidum
lucidum for 6 months.
synthesis and blocking of sterol receptors in GI tract owing to
structural similarity between lanosterol- derived terpenes and cholesterol. Further study needed, as only one human trial published to date.
214 T ABLE 2 Continued
Ref. CVD
Effect tested
Type of study
Reported effect
Comment
Platelet aggregation
110, 147–152 and blood
In vitro and
Adenosine, adenosine
No in vivo studies
animal
derivative, and
published to date.
clotting
studies
ganodermic acid S isolated from G. lucidum inhibited platelet aggregation; metalloprotease isolated from G. lucidum increased clotting time.
CVD Glycemic control
Animal studies
Polysaccharides isolated
Positive hypoglycemic
and human
from G. lucidum shown
results in animal
intervention
to have hypoglycemic
studies; however, there
trial
effect in mice. In a
is a lack of data from
small, poorly controlled
well-designed human
clinical trial, G. lucidum
trials to support this
supplementation was
claim.
reported to improve glycemic control in type
I (n = 2) and type II
Wachtel-Galor
(n = 2) diabetes mellitus patients, compared to 4 untreated control diabetic subjects.
Inflammation Inflammatory
32,156,158 et response
In vitro and
Anti-inflammatory
Interesting results;
animal
properties were shown
however, no human
al.
studies
in induced irritation
trials published to date,
Lingzhi
(edema) in mice after
and no data on
oral and topical
mechanism of anti-
application of organic
inflammatory effect of
(terpenes-containing)
triterpenes presented.
Polyphorous
extracts.
Inflammation PLA2 activity
In vitro
Some isolated triterpenes
Results indicate that
showed inhibitory
different triterpenes
effects against the
may be active only
enzyme PLA2 isolated
against certain types of
from bee venom, hog
PLA2. Specific
Fungus
pancreas, and snake
triterpenes should be
venom. One triterpene
further studied in
(ganoderic acid T)
specific models of
(Ganoderma
reported to inhibit
PLA2 inhibition and the
human recombinant
downstream
PLA2.
antiinflammatory effects of this.
Allergy Allergic response
In vitro and
Triterpenes from G.
No strong evidence of
human trials
lucidum reported to
antiallergenic
163–165 lucidum
have inhibited
properties; further
histamine release in
study needed.
vitro. No reports of
However, possible
allergic response to
allergic reaction to G.
ingestion of G. lucidum,
lucidum should be
but three reports of
considered, especially
aerosensitization in
in patients with
atopic subjects.
respiratory or allergic disorders or in association with topical application.
216 T ABLE 2 Continued
Ref. Antioxidant
Effect tested
Type of study
Reported effect
Comment
ROS scavenging
169–175 and antioxidant
In vitro and
G. lucidum found to
Not yet known which
human
possess antioxidant
antioxidants from G.
bioavailability
intervention
properties in in vitro
lucidum are absorbed or
trial
models; reported to
if the absorbed
scavenge superoxide
antioxidants have any
and hydroxyl radical, to
in vivo protective
protect DNA from ROS
effect. Further study
and decrease lipid
needed.
peroxidation. In a human bioavailability study (n = 10), plasma antioxidant power increased after G. lucidum intake.
Liver injury Induced liver injury
Animal studies
G. lucidum reported to
No histological data
show antifibrotic and
presented; whether the
hepatoprotective
effect of G lucidum is
effects after CCl4 or
truly protective,
ethanol in animal
indicates faster
Wachtel-Galor
models induced liver
recovery from damage,
injury.
or improved clearing of plasma biomarkers of liver injury should be further investigated.
a The studies summarized in this table are those published in English in at least abstract form. et al.
Lingzhi Polyphorous Fungus (Ganoderma lucidum) 217
often not been well defined in terms of the source, growing conditions, means of identification of the mushroom as G. lucidum, contamination, heavy metal content, batch-to-batch variation, method of extraction/preparation, or dosage. Most studies have been performed on animals or in cell culture models, and experimental studies have often been small, poorly designed, and inadequately controlled. The great wealth of chemical data and anecdotal evidence on effects of G. lucidum needs now to be complemented by reliable experimental and clinical data from human trials to clearly establish whether the reported health-related effects are valid and significant. Quality control procedures to define and standardize G. lucidum preparations, in addition to well-designed animal and cell culture studies, are needed to determine mechanisms of action and to help characterize the active component(s) of this putative medicinal mushroom.
ACKNOWLEDGMENTS The authors thank the Hong Kong Polytechnic University for funding this
work.
REFERENCES 1. Chang ST, Buswell JA. Mushroom nutriceuticals. World J Micro Biotechnol
1996; 12:473–476. 2. Rajarathnam S, Shashirekha MN, Bano Z. Biodegradative and biosynthetic capacities of mushrooms: present and future strategies. Cr Rev Biotechnol 1998; 18:91–236. 3. Borchers AT, Stern JS, Hackman RM, Keen CL, Gershwin ME. Minireview: mushrooms, tumors and immunity. Proc Soc Exp Biol Med 1999; 221:281– 293. 4. Wasser SP, Weis AL. Medicinal properties of substances occuring in higher basidiomycetes mushrooms: current perspectives. Int J Med Mushrooms 1999; 1:31–62. 5. Wasser SP, Weis AL. Therapeutic effects of substances occuring in higher basidiomycetes mushroom: a modern perspective. Cr Rev Immunol 1999; 19:65–96. 6. Jong SC, Birmingham JM. Medicinal benefits of the mushroom Ganoderma. Adv Appl Microbiol 1992; 37:101–134. 7. Shiao MS, Lee KR, Lin LJ, Wang CT. Natural products and biological activities of the Chinese medicinal fungus Ganoderma lucidum. Am Chem Soc Symp Series 1994; 547:342–354. 8. Chen AW, Miles PG. Biomedical research and the application of mushroom nutriceuticals from Ganoderma lucidum. In: Royse DJ, ed. Proceedings of the
218 Wachtel-Galor et al. 2nd International Conference on Mushroom Biology and Mushroom Products.
University Park: Penn State University, 1996:161–175. 9. Teeguarden R. The Ancient Wisdom of the Chinese Tonic Herbs. New York: Warner Books, 1998. 10. Chang ST, Buswell JA. Ganoderma lucidum (Curt.: Fr.) P. Karst. (Aphyllo- phoromycetideae)—a mushrooming medicinal mushroom. Int J Med Mush- rooms 1999; 1:139–146. 11. Upton R. American Herbal Pharmacopeia and Therapeutic Compendium:
Reishi Mushroom, Ganoderma lucidum. Standards of Analysis, Quality Con- trol, and Therapeutics. Santa Cruz, CA, 2000.
12. Willard T. Reishi Mushroom, Herb of Spiritual Potency and Medical Wonder. Issaquah, WA: Sylvan Press, 1990. 13. Teow SS. Effective dosage of the extract of Ganoderma lucidum in the treatment of various ailments. In: Royse DJ, ed. Proceedings of the 2nd International Conference on Mushroom Biology and Mushroom Products. Penn State Uni- versity, 1996:177–185. 14. Donk MA. A conspectus of the families of Aphyllophorales. Persoonia 1964; 3:19–24. 15. Moncalvo JM. Systematics of Ganoderma. In: Flood J, Bridge PD, Holderness M, eds. Ganoderma Diseases of Perennial Crops. Wallingford, UK: CAB International, 2000:23–45. 16. Mizuno T, Wang G, Zhang J, Kawagishi H, Nishitoba T, Reishi Li J. Ganoderma lucidum and Ganoderma tsugae: bioactive substances and medicinal effects. Food Rev Int 1995; 11:151–166. 17. Li Shizhen, Ben Cao Gang Mu, Revised ed. Beijing: Zhongguo Zhong yi yao chu ban she, 1998. Chinese. 18. Baxter AP, Eicker A. Preliminary synopsis: recorded taxa of Southern Africa Ganodermataceae. Proceedings of Contributed Symposium, 59A,B, 5th Inter- national Mycological Congress, Vancouver, Aug 14–21, 1994:3–5. 19. Zhao JD, Zhang XQ. Importance, distribution and taxonomy of Ganoderma- taceae in China. Proceedings of Contributed Symposium, 59A,B, 5th Interna- tional Mycological Congress, Vancouver, Aug 14–21, 1994:1–2. 20. Szedlay G, Jakucs E, Boldizsar I, Boka K. Basidiocarp and mycelium morphology of Ganoderma lucidum Karst. strains isolated in hungary. Acta Microbiol Immunol Hung 1999; 46:41–52. 21. Woo YA, Kim HJ, Cho JH, Chung H. Discrimination of herbal medicines according to geographical origin with near infrared reflectance spectroscopy and pattern recognition techniques. J Pharm Biomed Anal 1999; 21:407– 413. 22. Ryvarden L. Can we trust morphology in Ganoderma? In: Buchanan PK, Hseu RS, Moncalvo JM, eds. Ganoderma: Systematics, Phytopathology and Phar- macology. Proceedings of Contributed Symposium, 59A,B, 5th International Mycological Congress, Vancouver, Aug 14–21, 1994:19–24. 23. Gottlieb AM, Saidman BO, Wright JE. Isoenzymes of Ganoderma species from southern South America. Mycol Res 1998; 102:415–426.
Lingzhi Polyphorous Fungus (Ganoderma lucidum) 219 24. Hseu RS, Wang HH, Wang HF, Moncalvo JM. Differentiation and grouping of
isolates of Ganoderma lucidum complex by random amplified polymorphic DNA-PCR compared with grouping on the basis of internal transcribed spacer sequences. Appl Environ Microbiol 1996; 62:1354–1363. 25. Chen DH, Shiou WY, Wang KC, Huang SY, Shie YT, Tsai CM, Shie JF, Chen KD. Chemotaxonomy of triterpenoid pattern of HPLC of Ganoderma lucidum and Ganoderma tsugae. J Chinese Chem Soc 1999; 46:47–51. 26. Gottlieb AM, Ferref E, Wright JE. rDNA analyses as an aid to the taxonomy of species of Ganoderma. Mycol Res 2000; 104:1033–1045. 27. Su CH, Yang YZ, Ho HO, Hu CH, Sheu MT. High-performance liquid chro- matographic analysis for the characterization of triterpenoids from Ganoderma. J Chromatogr Sci 2001; 39:93–100. 28. Moncalvo JM, Wang HF, Hseu RS. Phylogenetic relationships in Ganoderma inferred from the internal transcribed spacers and 25s ribosomal DNA sequences. Mycologia 1995; 87:223–238. 29. Moncalvo JM, Wang HF, Hseu RS. Gene phylogeny of the Ganoderma lucidum complex: comparison with traditional taxonomic characters. Mycol Res 1995; 99:1489–1499. 30. Moncalvo JM, Wang HF, Wang HH, Hseu RS. The use of rDNA nucleotide sequence data for species identification and phylogeny in the Ganodermataceae. In: Buchanan PK, Hseu RS, Moncalvo JM, eds. Ganoderma: Systematics, Phytopathology and Pharmacology. Proceedings of Contributed Symposia 59A,B, 5th International Mycological Congress, Vancouver, Canada, August 1994:31–44. 31. Zhu YP. Chinese Materia Medica. Singapore: Harwood Academic Publishers, 1998. 32. Patocka J. Anti-inflammatory triterpenoids from mysterious mushroom Ganoderma lucidum and their potential possibility in modern medicine. Acta Med (Hradec Kralove) 1999; 42:123–125. 33. Compiled by the State Pharmacopoeia Commission of P.R. China Pharmaco- poeia of the People’s Republic of China. English ed. Beijing, China: Chemical Industry Press, 2000. 34. Mayzumi F, Okamoto H, Mizuno T. Cultivation of Reishi. Food Rev Int 1997; 13:365–373. 35. Riu H, Roig G, Sancho J. Production of carpophores of Lentinus edodes and Ganoderma lucidum grown on cork residues. Microbiologia SEM 1997; 13:185– 192. 36. Tseng TC, Shiao MS, Shieh YS, Hao YY. Studies on Ganoderma lucidum: liquid culture and chemical composition of mycelium. Bot Bull Acad Sin 1984; 25:149– 157. 37. Cha DY, Yoo YB. Cultivation techniques of Reishi. Food Rev Int 1997; 13:373–382. 38. Yang FC, Liau CB. The influence of environmental conditions on polysaccha- ride formation by Ganoderma lucidum in submerged cultures. Process Biochem 1998; 33:547–553.
220 Wachtel-Galor et al. 39. Lee KM, Lee SY, Lee HY. Bistage control of pH for improving exopolysac-
charide production from mycelia of Ganoderma lucidum in an air-lift Fermentor. J Bios Bioeng 1999; 88:646–650. 40. Habijanic J, Berovic M. The relevance of solid-state substrate moisturing on Ganoderma lucidum biomass cultivation. Food Technol Biotechnol 2000; 38:225–228. 41. Yang F, Ke Y, Kuo S. Effect of fatty acids on the mycelial growth and poly- saccharide formation by Ganoderma lucidum in shake flask cultures. Enzyme Microb Technol 2000; 27:295–301. 42. Fang QH, Zhong JJ. Two-stage culture process for improved production of ganoderic acid by liquid fermentation of higher fungus Ganoderma lucidum. Biotechnol Prog 2002; 18:51–54. 43. Kim SW, Hwang HJ, Park JP, Cho YJ, Song CH, Yun JW. Mycelial growth and exo-biopolymer production by submerged culture of various edible mushrooms under different media. Lett Appl Microbiol 2002; 34:56–60. 44. Chang J. Scientific evaluation of traditional Chinese medicine under DSHEA: a conundrum. J Altern Complem Med 1999; 5:181–189. 45. Chang R. Effective dose of Ganoderma in humans. Proceedings of Contributed Symposium, 59A,B, 5th International Mycological Congress, Vancouver, Aug 14–21, 1994:117–121. 46. Teow SS. The therapeutic value of Ganoderma lucidum. Proceedings of Con- tributed Symposium, 59A,B, 5th International Mycological Congress, Van- couver, Aug 14–21, 1994:105–113. 47. Kim MJ, Kim HW, Lee YS, Shim MJ. Studies on safety of Ganoderma lucidum [abstr]. Kor J Mycol 1986; 14:49–59. 48. Chiu SW, Wang ZM, Leung TM, Moore D. Nutritional value of Ganoderma extract and assessment of its genotoxicity and antigenotoxicity using comet assays of mouse lymphocytes. Food Chem Toxicol 2000; 38:173–178. 49. Mau JL, Lin HC, Chen CC. Non-volatile components of several medicinal mushrooms. Food Res Int 2001; 34:521–526. 50. Kim HS, Kacew S, Lee BM. In vitro chemopreventive effects of plant poly- saccharides (aloe Barbadensis miller, Lentinus endodes, Ganoderma lucidum and Coriolus versicolor ). Carcinogenesis 1999; 20:1637–1640. 51. Kim BK, Chung HS, Chung KS, Yang MS. Studies on the antineoplastic components of Korean basidiomycetes. Korean J Mycol 1980; 8:107–113.
52. Miyazaki T, Nishijima M. Studies on fungal polysaccharides. XXVII. Structural examination of water-soluble, antitumor polysaccharide of Gano- derma lucidum . Chem Pharm Bull 1981; 29:3611–3616.
53. Hyun JW, Choi EC, Kim BK. Studies on constituents of higher fungi of Korea (LXVII), antitumor components of the basidiocarp of Ganoderma lucidum. Korean J Mycol 1990; 18:58–69. 54. Bao X, Liu C, Fang J, Li X. Structural and immunological studies of a major polysaccharide from spores of Ganoderma lucidum (Fr.) Karst. Carbohydr Res 2001; 332:67–74. 55. Wang YY, Khoo KH, Chen ST, Lin CC, Wong CH, Lin CH. Studies on the
Lingzhi Polyphorous Fungus (Ganoderma lucidum) 221 immuno-modulating and antitumor activities of Ganoderma lucidum (Reishi)
polysaccharides: functional and proteomic analyses of a fucose-containing glycoprotein fraction responsible for the activities. Bioorg Med Chem 2002; 10:1057–1062. 56. Sone Y, Okuda R, Wada N, Kishida E, Misaki A. Structures and antitumor activities of the polysaccharides isolated from fruiting body and the growing culture of mycelium of Ganoderma lucidum. Agric Biol Chem 1985; 49:2641– 2653. 57. Tomoda M, Gonda R, Kasahara Y, Hikino H. Glycan structures of ganoderans
B and C, hypoglycemic glycans of Ganoderma lucidum fruit bodies. Phyto- chemistry 1986; 25:2817–2820. 58. Bao X, Duan J, Fang X, Fang J. Chemical modifications of (1!3)-a- D -glucan from spores of Ganoderma lucidum and investigation of their physiochem- ical properties and immunological activity. Carbohydr Res 2001; 336:127– 140. 59. Bao X, Wang X, Dong Q, Fang J, Li X. Structural features of immunologically active polysaccharides from Ganoderma lucidum. Phytochemistry 2002; 59:175– 181. 60. Eo SK, Kim YS, Lee CK, Han SS. Antiherpetic activities of various protein bound polysaccharides isolated from Ganoderma lucidum. J Ethnopharmacol 1999; 68:175–181. 61. Hikino H, Konno C, Mirin Y, Hayashi T. Isolation and hypoglycemic activity of ganoderans A and B, glycans of Ganoderma lucidum fruit bodies. Planta Medi 1985; 4:339–340. 62. Zhang L, Zhang M, Chen J. Solution properties of antitumor carboxymethy-
lated derivatives of a-(1!3)- D -glucan from Ganoderma lucidum. Chinese J Polymer Sci 2001; 19:283–289.
63. Bettelheim FA, March J. Introduction to General, Organic and Biochemistry. Orlando, FL: Harcourt Brace College Publishers, 1995. 64. Parker SP. McGraw-Hill Dictionary of Chemistry. New York: McGraw-Hill, 1997. 65. Das MC, Mahato SB. Triterpenoids. Phytochemistry 1983; 22:1071–1095. 66. Mahato SB, Sen S. Advances in triterpenoid research, 1990–1994. Phytochem- istry 1997; 44:1185–1236. 67. Safayhi H, Sailer ER. Anti-inflammatory actions of pentacyclic triterpenes. Planta Med 1997; 63:487–493. 68. Craig WJ. Health-promoting properties of common herbs. Am J Clin Nutr 1999; 70:491S–499S. 69. Mashour NK, Lin GI, Frishman WH. Herbal medicine for the treatment of cardiovascular disease: clinical considerations. Arch Intern Med 1998; 158:2225–2234. 70. Yun TK. Update from Asia—Asian studies on cancer chemoprevention. Ann NY Acad Sci 1999; 889:157–192. 71. Haralampidis K, Trojanowska M, Osbourn AE. Biosynthesis of triterpenoid saponins in plants. Adv Biochem Eng Biotechnol 2002; 75:31–49.
222 Wachtel-Galor et al. 72. Budavari S. The Merck Index. 11th ed. Rahway, New Jersey: Merck & Co.,
1989:845. 73. Abe I, Rohmer M, Prestwich GD. Enzymatic cyclization of squalene and oxidosqualene to sterols and triterpenes. Chem Rev 1993; 93:2189–2206. 74. Kubota T, Asaka Y, Miura I, Mori H. Structures of ganoderic acid A and B, two new lanostane type bitter triterpenes from Ganoderma lucidum (Fr.) Karst. Helv Chim Acta 1982; 65:611–619. 75. Nishitoba T, Sato H, Kasai T, Kawagishi H, Sakamura S. New bitter C27 and C30 terpenoids from fungus Ganoderma lucidum (Reishi). Agric Biol Chem 1984; 48:2905–2907. 76. Hirotani M, Furuya T, Shiro MA. Ganoderic acid derivative, a highly oxygenated lanostane type triterpenoid from Ganoderma lucidum. Phytochem- istry 1985; 24:2055–2059. 77. Kohda H, Tokumoto W, Sakamoto K, Fuji M, Hirai Y, Yamasaki K, Komoda Y, Nakamura H, Ishihara S, Uchida M. The biologically active constituents of Ganoderma lucidum (Fr.) Karst. histamin release-inhibitory triterpenes. Chem Pharm Bull 1985; 33:1367–1374. 78. Komoda Y, Nakamura H, Ishihara S, Uchida M, Kohda H, Yamasaki K. Structures of new terpenoid constituents of Ganoderma lucidum (Fr.) Karst (polyporaceae). Chem Pharm Bull 1985; 33:4829–4835. 79. Morigiwa A, Kitabatake K, Fujimoto Y, Ikekawa N. Angiotensin converting enzyme-inhibitory triterpenes from Ganoderma lucidum. Chem Pharm Bull 1986; 34:3025–3028. 80. Nishitoba T, Sato H, Sakamura S. New terpenoids, ganolucidic acid D, ganoderic acid L, lucidone C and lucidenic acid G, from the fungus Ganoderma lucidum . Agric Biol Chem 1986; 50:809–811. 81. Sato H, Nishitoba T, Shirasu S, Oda K, Sakamura S. Ganoderiol A and B, new triterpenoids from the fungus Ganoderma lucidum (Reishi). Agric Biol Chem 1986; 50:2887–2890. 82. Lin LJ, Shiao MS. Seven new triterpenes from Ganoderma lucidum. J Nat Prod 1988; 51:918–924. 83. Lin LJ, Shiao MS, Yeh SF. Triterpenes from Ganoderma lucidum. Phytochem- istry 1988; 27:2269–2271. 84. Min BS, Nakamura N, Miyashiro H, Bae K-W, Hattori M. Triterpenes from the spores of Ganoderma lucidum and their inhibitory activity against HIV-1 protease. Chem Pharm Bull 1998; 46:1607–1612. 85. Gonzalez AG, Leon F. Rivera A, Munoz CM, Bermejo J. Lanostanoid triter- penes from Ganoderma lucidum. J Nat Prod 1999; 62:1700–1701. 86. Mizushina Y, Takahashi N, Hanashima L, Koshino H, Esumi Y, Uzawa J, Sugawara F, Sakaguchi K. Lucidenic acid O and lactone, new terpene inhibitors of eukaryotic DNA polymerases from a basidiomycete, Ganoderma Lucidum. Bioorg Med Chem 1999; 7:2047–2052. 87. Ha TBT, Gerhauser C, Zhang WD, Ho NCL, Fouraste I. New lanostanoids from Ganoderma Lucidum that include NAD(P)H:quinone oxidoreductase in cultured Hepalclc7 murine hepatoma cells. Planta Med 2000; 66:681–684.
Lingzhi Polyphorous Fungus (Ganoderma lucidum) 223 88. Min BS, Gao JJ, Nakamura N, Hattori M. Triterpenes from the spores of
Ganoderma Lucidum and their cytotoxicity against Meth-A and LLC tumor cells. Chem Pharm Bull 2000; 48:1026–1033. 89. Wu TS, Shi LS, Kuo SC. Cytotoxicity of Ganoderma lucidum triterpenes. J Nat Prod 2001; 64:1121–1122. 90. Ma J, Ye Q, Hua Y, Zhang D, Cooper R, Chang MN, Chang JY, Sun HH. New lanostanoids from the mushroom Ganoderma lucidum. J Nat Prod 2002; 65:72–75. 91. Chen TQ, Li KB, He XJ, Zhu PG, Xu J. Micro-morphology, chemical compo- nents and identification of log-cultivated Ganoderma Lucidum spore. Proc ’98 Nanjing Intl Symp Science and Cultivation of Mushroom, 1998: 214. 92. Byrne AR, Ravnik V. Trace element concentrations in higher fungi. Sci Tot Envir 1976; 6:65–78. 93. Stijve T, Besson R. Mercury, cadmium, lead and selenium content of mushroom species belonging to the genus Agaricus. Chemosphere 1976; 2:151–158. 94. Thomas K. Heavy metals in urban fungi. Mycologist 1992; 6:195–196. 95. Mejstrik V, Lepsova A. Applicability of fungi to the monitoring of environ- mental pollution by heavy metals. In: Markert B, ed. Plant as Biomonitors. Weinheim: VCH, 1993:365–378. 96. Vetter J. Data on arsenic and cadmium contents of some common mushrooms. Toxicon 1994; 32:11–15. 97. Michelot D, Poirier F, Melendez-Howell LM. Metal content profiles in mushrooms collected in primary forests of Latin America. Arch Environ Contam Toxicol 1999; 36:256–263. 98. Tham LX, Matsuhashi S, Kume T. Responses of Ganoderma lucidum to heavy metals. Mycoscience 1999; 40:209–213. 99. Ybanez N, Montoro R. Trace element food toxicology: an old and ever-growing discipline. Cr Rev Food Sci Nutr 1996; 36:299–320.
100. Mino Y, Ota N, Sakao S, Shimomura S. Determination of germanium in medicinal plants by atomic absorption spectrometry with electrothermal atomization. Chem Pharm Bull 1980; 28:2687–2691.
101. Kolesnikova OP, Tuzova MN, Kozlov VA. Screening of immunoactive properties of alkanecarbonic acid derivatives and germanium-organic com- pounds in vivo. Immunologiya 1997; 10:36–38.
102. Schimmer O, Eschelbach H, Breitinger DK, Gruetzner K, Wick H. Organo- germanium compounds as inhibitors of the activity of direct acting mutagens in Salmonella typhimurium. Arzneim-Forsch 1997; 47:1398–1402.
103. Lee S, Park S, Oh JW, Yang CH. Natural inhibitors for protein prenyltransfer- ase. Planta Med 1998; 64:303–308. 104. Kino K, Yamashita A, Yamaoka K, Watanabe J, Tanaka S, Shimizu K, Tsunoo H. Isolation and characterization of a new immunomodulatory protein, Ling Zhi-8 (LZ-8), from Ganoderma lucidum. J Biol Chem 1989; 264:472–478.
105. Kino K, Sone T, Watanabe J, Yamashita A, Tsuboi H, Miyajima H, Tsunoo H. Immunomodulator, LZ-8, prevents antibody production in mice. Int J Immunopharmac 1991; 13:1109–1115.
224 Wachtel-Galor et al. 106. Van Der Hem L, Van Der Vliet A, Bocken CFM, Kino K, Hoitsma AJ, Tax
WJM. Ling Zhi-8: studies of a new immunomodulating agent. Transplantation 1995; 60:438–443.
107. Kawagishi H, Mitsunaga SI, Yamawaki M, Ido M, Shimada A, Kinoshita T, Murata T, Usui T, Kimura A, Chiba S. A lectin from mycella of the fungus Ganoderma lucidum . Phytochemistry 1997; 44:7–10.
108. Wang H, Ng TB, Ooi VEC. Lectins from mushrooms. Mycol Res 1998; 102:897–906. 109. Mizushina Y, Watanabe I, Togashi H, Hanashima L, Takemura M, Ohta K, Sugawara F, Koshino H, Esumi Y, Uzawa J, Matukage A, Yoshida S, Sakaguchi K. An ergosterol peroxide, a natural product that selectively enhances the inhibitory effect of linoleic acid on DNA polymerase h. Biol Pharm Bull 1998; 21:444–448.
110. Choi HS, Sa YS. Fibrinolytic and antithrombotic protease from Ganoderma lucidum . Mycologia 2000; 92:545–552. 111. Flint SJ, Enquist LW, Krug RM, Racaniello VR, Skalka AM. Principles of Virology: Molecular Biology, Pathogenesis, and Control. Washington, DC: ASM Press, 2000:683–712.
112. Eo SK, Kim YS, Lee CK, Han SS. Antiviral activities of various water and methanol soluble substances isolated from Ganoderma lucidum. J Ethno- pharmacol 1999; 68:129–136.
113. Kim YS, Eo SK, Oh KW, Lee CK, Han SS. Antiherpetic activities of acidic protein bound polysaccharide isolated from Ganoderma lucidum alone and in combinations with interferons. J Ethnopharmacol 2000; 72:451–458.
114. Oh KW, Lee CK, Kim YS, Eo SK, Han SS. Antiherpetic activities of acidic protein bound polysaccharide isolated from Ganoderma lucidum alone and in combination with acyclovir and vidarabine. J Ethnopharmacol 2000; 72:221– 227.
115. El-Mekkawy S, Meselhy MR, Nakamura N, Tezuka Y, Hattori M, Kakiuchi N, Shimotohno K, Kawahata T, Otake T. Anti-HIV-1 and anti-HIV-1-protease substances from Ganoderma lucidum. Phytochemistry 1998; 49:1651–1657.
116. Hijikata Y, Yamada S. Effect of Ganoderma lucidum on postherpetic neuralgia. Am J Chinese Med 1998; 26:375–381. 117. Weber R, Christen L, Loy M, Schaller S, Christen S, Joyce CRB, Ledermann U, Ledergerber B, Cone R, Luthy R, Cohen MR. Randomized, placebo-controlled trial of Chinese herb therapy for HIV-1-infected individuals. J Acquir Immune Defic Syndr 1999; 22:56–64.
118. Robbins WJ, Kavanagh F, Hervey A. Antibiotic substances from basidiomy- cetes I. Pleuro. Proc Natl Acad Sci USA 1947; 33:171–176. 119. Yoon SY, Eo SK, Kim YS, Lee CK, Han SS. Antimicrobial activity of Ganoderma lucidum extract alone and in combination with some antibiotics. Arch Pharm Res 1994; 17:438–442.
120. Nostro A, Germano MP, D’Angelo V, Marino A, Cannatelli MA. Extraction methods and bioautography for evaluation of medicinal plant antimicrobial activity. Lett Appl Microbiol 2000; 30:379–384.
Lingzhi Polyphorous Fungus (Ganoderma lucidum) 225 121. Ohno N, Miura NN, Sugawara N, Tokunaka K, Kirigaya N, Yadomae T.
Immunomodulation by hot water and ethanol extracts of Ganoderma lucidum. Pharm Pharmacol Lett 1998; 4:174–177.
122. Hong Kong Department of Health Annual Report 1999/2000 at www.info. gov.hk/dh/ar9900/ar9900.htm 123. World Health Report, mortality statistics, 2001. www.who.int 124. Chihara G, Hamuro J, Maeda YY, Arai Y, Fukuoka F. Fractionation and
purification of the polysaccharides with marked antitumor activity, especially lentinan, from Lentinus edodes (Berk.) Sing. (an edible mushroom). Cancer Res 1970; 30:2776–2781.
125. Wang G, Zhang J, Mizuno T, Zhuang C, Ito H, Mayuzumi H, Hidehumi O, Li J. Antitumor active polysaccharides from the Chinese mushroom Songshan Lingzhi, the fruiting body of Ganoderma tsugae. Biosci Biotechnol Biochem 1993; 57:894–900.
126. Zhang QH, Lin ZB. The antitumor activity of Ganoderma lucidum (Curt.:Fr.) P. Karst. (Ling Zhi) (Aphyllophoromycetideae) polysaccharides is related to tumor necrosis factor-a and interferon-g. Int J Med Mushrooms 1999; 1:207– 215.
127. Furusawa E, Chou SC, Furusawa S, Hirazumi A, Dang Y. Antitumour activity of Ganoderma lucidum, an edible mushroom, on intraperitoneally implanted Lewis lung carcinoma in synergenic mice. Phytother Res 1992; 6:300–304.
128. Lu H, Uesaka T, Katoh O, Kyo E, Watanabe H. Prevention of the development of preneoplastic lesions, aberrant crypt foci, by a water-soluble extract from cultured medium of Ganoderma lucidum (Rei-shi) mycelia in male F344 rats. Oncol Rep 2001; 8:1341–1345.
129. Wang SY, Hsu ML, Hsu HC, Tzeng CH, Lee SS, Shiao MS, Ho CK. The antitumor effect of Ganoderma lucidum is mediated by cytokines released from activated macrophages and Tlymphocytes. Int J Cancer 1997; 70:699–705.
130. Lieu CW, Lee SS, Wang SY. The effect of Ganoderma lucidum on induction of differentiation in leukemic U937 cells. Anticancer Res 1992; 12:1211–1216. 131. Lei LS, Lin ZB. Effect of Ganoderma polysaccharides on Tcell subpopulation and production of interlukin 2 in mixed lymphocyte response. Acta Pharm Sin 1992; 27:331–335.
132. Jia YF, Li H, Mori M, Zhang LX. Tumor necrosis factor production in mouse macrophages and its pharmacological modulation by Ganoderma lucidum. J Chinese Pharm Sci 1998; 7:29–32.
133. Szeto YT, Benzie IFF. Effects of dietary antioxidants on human DNA ex vivo. Free Rad Res 2002; 36:113–118. 134. Zhu HS, Yang XL, Wang LB, Zhao DX, Chen L. Effects of extracts from sporoderm-broken spores of Ganoderma lucidum on HeLa cells. Cell Biol Toxicol 2000; 16:201–206.
135. Mizushina Y, Hanashima L, Yamaguchi T, Takemura M, Sugawara F, Saneyoshi M, Matsukage A, Yoshida S, Sakaguchi K. A mushroom fruiting body-inducing substance inhibits activities of replicative DNA polymerases. Biochem Biophys Res Commun 1998; 249:17–22.
226 Wachtel-Galor et al. 136. Ramgolam V, Ang SG, Lai YH, Loh CS, Yap HK. Traditional Chinese medi-
cines as immunosuppressive agents. Ann Acad Med, Singapore 2000; 29:11–16. 137. Xia D, Li R, Lin Z, He Y. Effects of Ganoderma polysaccharides on immune function in mice. J Beij Med Univ 1989; 21:533–537. 138. Chen WC, Hau DM, Wang CC, Lin IH, Lee SS. Effects of Ganoderma lucidum and Krestin on subset T-cell in spleen of g-irradiated mice. Am J Chinese Med 1995; 23:289–298.
139. Miyasaka N, Inoue H, Totsuka T, Koike R, Kino K, Tsunoo H. An Immuno- modulatory protein, Ling zhi-8, facilitates cellular interaction through modu- lation of adhesion molecules. Biochem Biophy Res Commun 1992; 186:385– 390.
140. Kim RS, Kim HW, Kim BK. Suppressive effects of Ganoderma lucidum on proliferation of peripheral blood mononuclear cells. Mol Cells 1997; 7:52–55. 141. Wood D, De Backer G, Fargeman O, Graham I, Mancia G, Pyorala K, together with members of the Task Force. Prevention of coronary heart disease in clinical practice. Recommendations of the second joint Task Force of European and other societies on coronary prevention. Eur Heart J 1998; 19:1434–1503.
142. Liu KC, Phounsavan SF, Huang RL, Liao C, Hsu SY, Wang KJ. Phar- macological and liver functional studies on mycelium of Ganoderma lucidum. Chinese Pharm J 1988; 40:21–29.
143. Lee SY, Rhee HM. Cardiovascular effects of mycelium extract of Ganoderma lucidum : inhibition of sympathetic outflow as a mechanism of its hypotensive action. Chem Pharm Bull 1990; 38:1359–1364.
144. Kanmatsuse K, Kajiwara N, Hayashi K, Shimogaichi S, Fukinbara I, Ishikawa H, Tamura T. Studies on Ganoderma lucidum: efficacy against hypertension and side effects. Yakugaku Zasshi 1985; 105:942–947. Japanese. 145. Komoda Y, Shimizu M, Sonoda Y, Sato Y. Ganoderic acid and its derivatives
as cholesterol synthesis inhibitors. Chem Pharm Bull 1989; 37:531–533. 146. Dogne JM, Leval XX, Benoit P, Delarge J, Masereel B, David JL. Recent advances in antiplatelet agents. Curr Med Chem 2002; 9:577–589. 147. Shimizu A, Yano T, Saito Y, Inada Y. Isolation of an inhibitor of platelet aggregation from a fungus, Ganoderma lucidum. Chem Pharm Bull 1985; 33:3012–3015.
148. Kawagishi H, Fukuhara F, Sazuka M, Kawashima A, Mitsubori T, Tomita T. 5V-Deoxy-5V-methylsulphinyladenosine, a platelet aggregation inhibitor from Ganoderma lucidum . Phytochemistry 1993; 32:239–241.
149. Su CY, Shiao MS, Wang CT. Differential effects of ganodermic acid S on the thromboxane A2-signaling pathways in human platelets. Biochem Pharmacol 1999; 58:587–595.
150. Su CY, Shiao MS, Wang CT. Predominant inhibition of ganodermic acid S on the thromboxane A2-dependent pathway in human platelet response to collagen. Biochim Biophys Acta 1999; 1437:223–234.
151. Su CY, Shiao MS, Wang CT. Potentiation of ganodermic acid S on pros- taglandin E1-induced cyclic AMP elevation in human platelets. Thromb Res 2000; 99:135–145.
Lingzhi Polyphorous Fungus (Ganoderma lucidum) 227 152. Halushka PV. Thromboxane A2 receptors: where have you gone? Prostagland
Other Lipid Media 2000; 60:175–189. 153. Expert panel on detection, evaluation, and treatment of high blood cholesterol in adults (adult treatment panel III). Executive summary of the third report of the national cholesterol education program (NCEP). JAMA 2001; 285:2486– 2497.
154. Hikino H, Ishiyama M, Suzuki Y, Konno C. Mechanisms of hypoglycemic activity of ganoderan B: a glycan of Ganoderma lucidum fruit body. Planta Med 1989; 55:423–428.
155. Libby P, Ridker P, Maseri A. Inflammation and atherosclerosis. Circulation 2002; 105:1135–1143. 156. Streeper RT, Satsangi N. Anti-inflammatory components of mushroom extract
on SUPELCOSIL ABZ+Plus HPLC columns. SUPELCO 1995; 14:1–3. 157. Stevens CD. Clinical Immunology and Serology. Philadelphia: F.A. Davis Company, 1996:166–167. 158. Giner-Larza EM, Manez S, Giner-Pons RM, Recio MC, Rios JL. On the anti- inflammatory and anti-phospholipase A2 activity of extracts from lanostane- rich species. J Ethnopharmacol 2000; 73:61–69.
159. Zollner H. Handbook of Enzyme Inhibitors. 3d ed. Weinheim, New York, Chichaster, Brisbane, Singapore, Toronto: Wiley-VCH, 1999. 160. Dan P, Dagan A, Krimsky M, Pruzanski W, Vadas P, Yedgar S. Inhibition of type I and type II phospholipase A2 by phosphatidyl-ethanolamine linked to polymeric carriers. Biochemistry 1998; 37:6199–6204.
161. Jain MK, Yu BZ, Rogers JM, Smith AE, Boger ET, Ostrander RL, Rheingold AL. Specific competitive inhibitor of secreted phospholipase A2 from berries of Schinus terebinthifolius . Phytochemistry 1995; 39:537–547.
162. Wachtel-Galor S, Szeto YT, Ezioni S, Buswell JA, Neeman I, Benzie IFF. Antioxidant and anti-inflammatory properties of the lingzhi mushroom. 5th Annual Nutrition Symposium, Hong Kong, Nov 10, 2001.
163. Singh AB, Gupta SK, Pereira BMJ, Prakash D. Sensitization to Ganoderma lucidum in patients with respiratory allergy in India. Clin Exp Allergy 1995; 25:440–447.
164. Cutten AEC, Hasnain SM, Segedin BP, Bai TR, McKay EJ. The basidiomycete Ganoderma and asthma: collection, quantitation and immunogenicity of the spores. NZ Med J 1988; 101:361–363.
165. Gupta SK, Pereira BMJ, Singh AB. Ganoderma lucidum: partial character- ization of spore and whole body antigenic extracts. Invest Allergol Clin Immunol 2000; 10:83–89.
166. Thomas MJ. The role of free radicals and antioxidants: how do we know that they are working? Cr Rev Food Sci Nutr 1995; 35:21–39. 167. Strain JJ, Benzie IFF. Diet and antioxidant defence. In: Sadler M, Strain JJ, Cabellero B, eds. The Encyclopedia of Human Nutrition. London: Academic Press, 1999:95–105.
168. Young IS, Woodside JV. Antioxidants in health and disease. J Clin Pathol 2001; 54:176–186.
228 Wachtel-Galor et al. 169. Kim KC, Kim IG. Ganoderma lucidum extract protects DNA from strand
breakage caused by hydroxyl radical and UV irradiation. Int J Mol Med 1999; 4:273–277.
170. Lee JM, Kwon H, Jeong H, Lee JW, Lee SY, Baek SJ, Surh YJ. Inhibition of lipid peroxidation and oxidative DNA damage by Ganoderma lucidum. Phytother Res 2001; 15:245–249.
171. Shi Y, James AE, Benzie IFF, Buswell JA. Mushroom derived preparations in the prevention of H 2 O 2 -induced oxidative damage to cellular DNA. Teratog Carcinog Mutagen 2002; 22:103–111. 172. Zhu M, Chang Q, Wong LK, Chong FS, Li RC. Triterpene antioxidants from Ganoderma lucidum . Phytother Res 1999; 13:529–531. 173. Benzie IFF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of ‘‘antioxidant power’’: the FRAP assay. Anal Biochem 1996; 239:70– 76.
174. Benzie IFF, Szeto YT. Total antioxidant capacity of teas by the ferric reducing/ antioxidant power assay. J Agric Food Chem 1999; 47:633–636. 175. Wachtel-Galor S, Szeto YT, Tomlinson B, Benzie IFF. Ganoderma lucidum (‘Lingzhi’): acute and short-term biomarker response to supplementation. Int J Food Sci Nutr 2004; 55(1):75–83.
176. Lin JM, Lin CC, Chen MF, Ujiie T, Takada A. Radical scavenger and anti- hepatotoxic activity of Ganoderma formosanum, Ganoderma lucidum and Gano- derma neo -japonicum. J Ethnopharmacol 1995; 47:33–41.
177. Kim DH, Shim SB, Kim NJ, Jang IS. h- Glucuronidase- inhibitory activity and hepatoprotective effect of Ganoderma lucidum. Biol Pharm Bull 1999; 22:162– 164.
178. Shieh YH, Liu CF, Huang YK, Yang JY, Wu IL, Lin CH, Lin SC. Evaluation of the hepatic and renal protective effects of Ganoderma lucidum in mice. Am J Chin Med 2001; 29:501–507.
179. Song CH, Yang BK, Ra KS, Shon DH, Park EJ, Go GI, Kim YH. Hepa- toprotective effect of extracellular polymer produced by submerged culture of Ganoderma lucidum WK-003. J Microbiol Biotechnol 1998; 8:277–279.
180. Park EJ, Ko G, Kim J, Dong HS. Antifibrotic effects of a polysaccharide extracted from Ganoderma lucidum, glycyrrhizin, and pentoxifylline in rats with cirrhosis induced by biliary obstruction. Biol Pharm Bull 1997; 20:417–420.
181. US Patent and Trademark Office web site: WWW.USPTO.GOV.
Epimedium Species
Sook Peng Yap and Eu Leong Yong National University of Singapore
Singapore, Republic of Singapore
I. INTRODUCTION Herba Epimedii, Berberidaceae (Chinese name ‘‘yinyanghuo,’’ or ‘‘horny
goat weed’’) is the dried aerial part of several species of Epimedium, including
E. sagittatum (Sieb. Et Zucc), E. koreanum Nakai, E. pubescens Maxim., E. wushanense T.S. Ying, and E. brevicornum Maxim (1). Epimedium is a woody, evergreen perennial indigenous to shady mountain areas in temperate to subtropical Asia. It has broad heart-shaped leaves, and comprises over 40 species throughout the world. Most species of the genus have been used in folk medicine as yang tonic, mainly in China, Japan, and Korea. It is pungent and sweet in flavor.
A. Processing Epimedium herb is collected in summer and fall when foliage is growing
luxuriantly. The leaves are collected and removed from the thick stalks and foreign matter, then dried in the sun or in the shade.
230 Yap and Yong
B. Preparation and Dosage Epimedium herb alone can be taken in the form of water-boiling extract or it
can be macerated in wine for an oral infusion. It is also used together with prepared rehmania root, curculigo rhizome, dogwood fruit, and wolfberry fruit. It is often found as one of the herbs used in various proprietary prod- ucts. The dosage of Epimedium herb for human consumption is in the range of 6–15 g, up to 30 g for a single use.
C. Traditional Use Epimedium is one of the most popular and effective kidney yang tonics of
Chinese herbalism. Its first recorded use dates back to the ancient text Shen Nong Ben Cao Jing (ca. 200 B.C. – A.D. 100). The herb is listed in the Phar- macopoeia of the People’s Republic of China (1) as having action to ‘‘reinforce the kidney yang, strengthen tendons and bones, and relieve rheumatic conditions’’ and indicated ‘‘for impotence, seminal emission, weakness of the limbs, rheumatoid arthralgia with numbness and muscle contracture, and climacteric hypertension.’’
It received its colloquial name ‘‘horny goat weed’’ when goats grazing on the herb were observed to have excessive copulating. Oriental populations
consider Epimedium an aphrodisiac and the herb is used to treat sexual dysfunction in both men and women.
In addition, herba Epimedii have been reputed to be effective in ther- apy for infertility, chronic nephritis, leukopenia, neurasthenia, asthenia, am- nesia, or corresponding symptoms in China and Japan for over 2000 years (2,3).
D. Modern Use In recent years, Epimedium has been extensively used for coronary diseases,
hypertension, hyperlipidemia, hepatitis B, immunomodulation, as well as to alleviate menopausal discomfort (4). The herb is also thought to have anti- cancer and antiaging properties. It has been the subject of a number of animal and human studies to explore its medicinal values.
II. DETERMINATION OF EPIMEDIUM SPECIES Classically, the species of Epimedium herb can be determined according to the
spur length of its flower, and through morphological, histological, and che- motaxonomical inspection. In many cases, these methods are unreliable, as hybridization between species occurs, resulting in diversification of popula- tions of Epimedium species.
Epimedium Species 231
A. Genetic Characterization In plant genomes, genes for the ribosomal RNA (rDNA) are normally clus-
tered in an array of multiple tandemly repeated copies of the cistron of 18S-ITS1-5.8S-ITS2-28S (5). The coding regions of 18S, 5.8S, and 28S rDNA sequence are highly conserved, whereas sequence homology within the ITS1 and ITS2 regions is much lower across the plant kingdom. Based on these features of the plant rDNA, molecular markers, such as random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP), are good candidates for the identification and authentication of plant species. RAPD and PCR-RFLP have been applied to explore the DNA, generate discrete and species-specific RFLP patterns, and hence, confirm the Epimedium species. E. sagittatum, E. koreanum, E. pubescens, and E. wushanense were easily distinguished by a representative amplified band pattern or DNA sequences (6,7). RAPD analysis is more appropriate than RFLP analysis for the determination of species because it can be conducted by an automated procedure rapidly and no enzymatic digestion is needed; thus less plant material is required.