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.

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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.