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Antioxidant and immunomodulatoryeffects of a α-glucan from fruit body ofmaitake (Grifola frondosa)Lei Honga, Wang Weiyub, Wang Qina, Guo Shuzhena & Wu lebinb

a School of Biology and Food Engineering, Hefei University ofTechnology, Hefei 230009, Chinab Shandong University Medical College, Shandong Medical ImagingResearch Institute, Jinan 250021, ChinaPublished online: 18 Jul 2012.

To cite this article: Lei Hong, Wang Weiyu, Wang Qin, Guo Shuzhen & Wu lebin (2013) Antioxidantand immunomodulatory effects of a α-glucan from fruit body of maitake (Grifola frondosa), Foodand Agricultural Immunology, 24:4, 409-418, DOI: 10.1080/09540105.2012.704901

To link to this article: http://dx.doi.org/10.1080/09540105.2012.704901

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Antioxidant and immunomodulatory effects of a a-glucan from fruitbody of maitake (Grifola frondosa)

Lei Honga*, Wang Weiyub, Wang Qina, Guo Shuzhena and Wu lebinb*

aSchool of Biology and Food Engineering, Hefei University of Technology, Hefei 230009, China;bShandong University Medical College, Shandong Medical Imaging Research Institute, Jinan250021, China

(Received 21 March 2012; final version received 18 June 2012)

The antioxidant and immunomodulatory effect of an a-glucan (designated here asMT-a-glucan) from fruit body of Maitake mushroom (Grifola frondosa) onD-galactose (D-gal)-induced senescent mice model was evaluated. Results showedthat MT-a-glucan could ameliorate age-related alternations in both motorand memory activities of senescent mice. Treatment with MT-a-glucan (450or 150 mg kg�1) significantly increased the body weight, hepatic superoxidedismutase and glutathione peroxidase activity, reduced glutathione content,the proliferative response and interleukine-2 (IL-2) production of splenocytesinduced by ConA. Treatment with MT-a-glucan significantly decreased hepaticmalondialdehyde content, the levels of macrophage proliferative response andIL-1 and NO production. These data suggest that MT-a-glucan has antioxidativeand immunomodulatory effects on D-gal-induced senescent mice.

Keywords: MT-a-glucan; ageing; D-galactose; antioxidation; immunomodulation

Introduction

Maitake mushrooms (Grifola frondosa) belong to Basidiomycetes in fungi and

have been praised and consumed by Chinese people for hundreds of years because

of their enticing taste and nutrition. It is well known in Chinese folks that eating

maitake mushrooms has a delaying-senescence effect. Moreover, the medicinal

properties of maitake have been claimed for years and some of them have been

demonstrated scientifically and experimentally. Its active parts are various poly-

saccharides derived from maitake mushrooms. For instance, polysaccharides derived

from maitake mushrooms have been shown to have antitumour effect (Liu, Chen, &

Wu, 2005), immune regulatory activity (Inoue, Kodoma, & Nanba, 2002), anti-

hyperliposis (Kubo, 1997), anti-common and specific infection effects such as

hepatitis (Kubo & Nanba, 1998; Ooi, 1996) and AIDS/HIV (Nanba, Kodama,

Schar, & Turner, 2000).

Previous studies showed that polysaccharides derived from maitake mushrooms

possess antioxidative properties in vitro (Li, Rong, & Wu, 2003) and immune

regulatory activity (Inoue et al., 2002). However, it is unknown whether poly-

*Corresponding authors. Email: leihong56@163.com; cjr.wulebin@vip.163.comLei Hong and Wang Weiyu contributed equally to this work.

Food and Agricultural Immunology, 2013

Vol. 24, No. 4, 409�418, http://dx.doi.org/10.1080/09540105.2012.704901

# 2012 Taylor & Francis

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saccharides derived from maitake mushrooms have an anti-ageing effect and its

mechanism of action concerning oxidative stress and immune response. On the basis

of previous studies, a new kind of a-glucan extracted and purified from the fruit

body of maitake, designed here as MT-a-glucan, was prepared in our laboratory

(Lei, Ma, & Wu, 2007). The present study was therefore designed to investigate the

anti-ageing effect of MT-a-glucan and its relation with its antioxidative and

immunomodulatory effect, exploiting D-galactose (D-gal)-induced senescent mice.

Materials and methods

Preparation of MT-a-glucan

MT-a-glucan was extracted and purified from fruit body of maitake (G. frondosa)

as previously described (Lei et al., 2007), which is a basically homogeneous

polysaccharide fraction and its molecular weight is about 400,000�450,000 Da.

MT-a-glucan was dissolved in 1% sodium carboxymethylcellulose (CMC-Na) and

diluted to the concentration needed.

Animals

Healthy Kunming strain mice and C57BL/6J mice, weighing 2092 g were supplied by

the Experimental Animal Center of Anhui Medical University (Hefei, China). They

were housed in plastic cages and maintained under standard conditions (12-h light�dark cycle; 23�258C; 35�60% r.h.). Before and during the experiment, mice were fed

with a normal laboratory pellet diet and water was freely available. After

randomisation into various groups, the mice were acclimatised in the new

environment for two days before initiation of the experiment. The study complied

with the current ethical regulations for the care and use of laboratory animals of

Hefei University of Technology (Anhui, China), and all mice used in the experiment

received humane care.

Main reagents

D-gal (Sigma Co.) was dissolved in 0.9% sterile saline at a concentration of 0.5% and

stored at 48C. Vitamin E (VE, Merck Co.) was dissolved in 1% CMC-Na with

absolute ethanol (less than 0.2%, v/v). Various measuring kits were used during the

study: superoxide dismutase (SOD) measurement kit, glutathione peroxidase

(GSHpx), reduced glutathione (GSH) and malondialdehyde (MDA) measurement

kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Concanavalin A

(ConA) and lipopolysaccharides (LPS) were obtained from Sigma; RPMI 1640

medium was obtained from Gibco which was supplemented with HEPES buffer

25 mmol L�1, sodium pyruvate 1 mmol L�1, L-glutamine 2 mmol L�1, penicillin

100 kU L�1, streptomycin 100 mg L�1 and 10% new born bovine serum and were

adjusted to pH 7.2. Bovine serum was supplied by Hangzhou Sijiqing Bioengineering

Co. (Hangzhou, China). All the other biochemicals and chemicals used in the

experiment were of analytical grade.

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Experimental design

In the experiment a total of 50 mice were used and were divided into 5 groups, each

containing 10 mice as follows: normal control; D-gal model control; 3 treatment groups

(given MT-a-glucan 450, 150 mg kg�1 or VE 50 mg kg�1). D-gal (40 mg kg�1) was

injected subcutaneously every other day for four weeks to induce senescent mice model

(Wang, Sun, Zhang, Jiang, & Zhang, 2002). Mice were given drugs or dissolvent

0.1 mL/10 g orally by gavage once a day for four weeks, followed by body weightmeasured each week.

Rotating-rod test

The rota-rod apparatus consisted of a base, two poles and a rod. The rod is 60 cmlong, 25 cm away from the base supported by two poles. At the beginning of the test,

put the mice on the rod slightly. Then the rod is revolved by hand at the rotating

speed of 10 rpm for one minute. The falling number of mice was recorded and the

falling percent was calculated (Lei et al., 2003).

Step-down-type passive avoidance test

The apparatus consisted of an acrylic box with a stainless-steel grid floor. A wooden

platform was fixed in the centre of the box. Electric shocks (40 mv) were delivered to

the grid floor for three seconds with an isolated pulse stimulator. At the beginning of

training, mice were placed in the box to adapt for three minutes. Then put the mice

on the platform slightly. When the mice step down and place all its paws on the grid

floor, it would jump to the platform as shock happened to be delivered. Step-downlatency (time of staying on the platform, SDL) and the number of errors (NOE) were

recorded within five minutes and repeated 24 h after training (Lei et al., 2003).

Biochemical measurements

At the end of the four-week treatment, mice were deprived of food overnight and

killed by decapitation. The liver homogenate was used for estimation of the levels of

SOD, GSHpx, GSH and MDA. The above biochemical parameters were determined

using commercial kits according to the guidelines indicated.

Proliferative response of splenocytes

The splenocytes suspension (1�1010 L�1) was prepared in a general way.

Splenocytes suspension 100 mL was seeded on a 96-well microtiter plate in the

presence of ConA (final concentration 3 mg L�1). The splenocytes were incubated at

378C in 5% CO2 atmosphere for 48 h. The supernatant, 150 mL, was collected and

stored at 208C until tested for interleukine-2 (IL-2) activity. Then RPMI-1640medium containing 150 mL 10% fetal calf serum (FCS) and 20 mL MTT (5 mg/mL)

was added to each well and splenocytes were cultured for another six hours. Then the

supernatant was discarded and 150 mL DMSO was added to each well. After shaking

the plate gently, the optical density (OD) values were measured at 570 nm (Song

et al., 2007). The results were expressed as means of OD value of triplicate wells.

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IL-2 assay

The supernatant containing IL-2 was diluted by 40 times. Activated splenocytes

suspension (2�109 L�1), 100 mL, was seeded on a 96-well microtiter plate in the

presence of the 100 mL dilution. The splenocytes were incubated at 378C in 5% CO2

atmosphere for 24 h (Song et al., 2007). MTT assay was done according to the

methods stated above. The results were expressed as means of OD value of triplicate

wells.

Proliferative response of macrophages and NO, NOS and iNOS assay

Mouse peritoneal macrophages were collected by lavage with cold D-Hank’s solution

(pH 7.4) and suspended in RPMI-1640 medium containing 10% FCS. One millilitre

cell suspension (2�109 L�1) was seeded in a 24-well microtiter plate and was

cultured at 378C in 5% CO2 atmosphere for two hours. After removing the culture

supernatant, the adherent cells were washed with 378C PBS and were cultured in

RPMI-1640 medium containing 10% FCS in the presence of LPS (final concentra-tion 6 mg L�1) at 378C in 5% CO2 atmosphere for six hours. Then the cells were

washed again with PBS and adherent cells were cultured in RPMI-1640 medium

containing 10% FCS at 378C in 5% CO2 atmosphere for 42 h (Andras, Gyongyi,

Laszlo, Tamas, & Gyorgy, 2006). The supernatant in each well was collected and was

used for NO, NOS and iNOS assay and IL-1 assay. NO, NOS and iNOS assays were

determined by using commercial kits according to the guidelines indicated. Cell

activity was assayed by MTT method according to the methods stated above. The

results were expressed as means of OD value of triplicate wells.

IL-1 assay

The supernatant containing IL-1 was diluted by 30 times. Mouse thymocytes were

prepared in a general way which was used for measuring IL-1 activity. Thymocytes

suspension (2�1010 L�1), 50 mL, was seeded in a 96-well microtiter plate in the

presence of 100 mL dilution and ConA (final concentration 3 mg L�1). The

thymocytes were incubated at 378C in a 5% CO2 incubator for 48 h (Song et al.,

2007). Cell activity was assayed by MTT method according to the methods statedabove. The results were expressed as means of OD value of triplicate wells.

Statistical analysis

Data were expressed as means9s.d. Statistical analysis was evaluated by one-way

analysis of variance, followed by the Student�Newman�Keuls test for multiple

comparisons, which was used to evaluate the difference between two groups.

P B0.05 was considered significant.

Results and discussion

Effect of MT-a-glucan on the body weight of of D-gal-induced ageing mice

The procedure of preparation of MT-a-glucan was studied in our laboratory.

The purity of the compound estimated by high performance gel permeation

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chromatography (HPGPC) demonstrated that the molecule was basically homo-

geneous, which had the molecular weight about 400,000�450,000 Da. Results of

structural analyses (IR, 1H-NMR and 13C-NMR) and monosaccharide analysis

(TLC and GC) demonstrated that the molecule is an a-glucan (Lei et al., 2007),

rather than a b-glucan, molecules hitherto reported to be most commonly produced

by this mushroom strain (Kubo, 1994, 1997). So this molecule is unique to maitake

among mushrooms according to the references we have searched. Another five

groups of polysaccharides having diverse molecular mass (470�1650 kDa) were

prepared from mycelium extract and submerged culture of G. frondosa, which had

antioxidant and free radical scavenging activities (Lee et al., 2003).Previous studies showed that chronic injections of D-gal subcutaneously into

mice could induce changes which resembled accelerated ageing (Wang et al., 2002).

The ageing model shows neurological impairment, decreased activity of antioxidant

enzymes and poor immune responses (Zhang & Li, 1990). Thus, we exploited

D-gal-treated mice as experimental senile model to investigate the anti-ageing effect

of MT-a-glucan and its mechanism of action. Results showed that all D-gal-treated

mice had the signs of senescent manifestation, such as low body weight, hair pull-off,

slow behaviour and lassitude. The success percentage of senescent mice model was 100%.

Figure 1 shows the effect of MT-a-glucan on body weight of D-gal-treated mice.

The body weight of D-gal model mice was much lower than that of the normal

control. After four weeks of treatment, MT-a-glucan (450, 150 mg kg�1) markedly

increased the body weight of D-gal-treated mice. This suggested that MT-a-glucan

could alleviate the lowered body weight of mice induced by D-gal.

Effects of MT-a-glucan on the behaviour of D-gal-induced ageing mice

The motor function and memory would decline with age. This might be related to the

declining of advanced function of the brain (Zhan, Liu, & Zhou, 1990). In this study,

step-down-type avoidance and rotating-rod tests were used to examine the long-term

memory and motor function, which are as index of senescence of mice.

Table 1 shows that D-gal (40 mg kg�1, sc every other day for four weeks)

increased the falling rate of mice determined by rotating-rod test (PB0.01),

shortened SDL (PB0.01) and increased NOE (PB0.01) determined by step-down

Figure 1. Effect of MT-a-glucan on body weight of D-gal-induced ageing mice.

Food and Agricultural Immunology 413

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test. The results of our study showed that the functions of motor and memory of

D-gal-treated mice were all lower than that of the normal young mice. MT-a-glucan

(450, 150 mg kg�1, ig once a day for four weeks) and VE (50 mg kg�1) markedly

improved motor and memory dysfunctions of D-gal-treated mice (Table 1). This

suggested that MT-a-glucan could improve equilibrium ability and brain function of

aged mice induced by D-gal, which resulted in delay in senility.

Effect of MT-a-glucan on levels of liver SOD, GSHpx, GSH and MDA inD-gal-induced ageing mice

There are several assumptions concerning ageing, of which the well-known theories

are free radical theory and immune theory. The free radical theory states that oxygen

free radicals are the important factors involved in the phenomenon of biological

ageing (Nohl, 1993). The immune theory states that the decreased immune function,

especially cellular immune function, is the main reason of ageing (Miller, 1991), in

which the most obvious change is the decrease of IL-2) production and IL-2 receptor

expression (Duan, Wang, & Shen, 1993). It has been demonstrated that poly-

saccharides derived from maitake mushrooms possess antioxidative properties in

vitro (Li et al., 2003) and immune regulatory activity (Inoue et al., 2002). In this

study, the anti-ageing effect of MT-a-glucan and its mechanism of action concerning

oxidative stress and immune response were investigated, exploiting D-gal-induced

senile mice model.

Table 1. Effect of MT-a-glucan on the behaviour of D-gal-induced ageing mice.

Group Dose (mg kg�1) Fall number Fall rate (%) SDL (s) NOE

Normal * 1** 10 87.11915.19** 0.9590.50**

D-gal � 8 80 49.35913.20 2.1390.77

MT-a-glucan 150 3** 30 70.52915.61** 0.9290.55**

450 1** 10 79.47913.45** 0.9090.49**

VE 50 3** 30 71.69914.24** 0.9790.62**

Note: Data are the mean9 s.d., n�10. In each vertical column, *P B0.05 and **P B0.01, comparedwith the D-gal group (analysis of variance followed by the Student�Newman�Keuls test).

Table 2. Effect of MT-a-glucan on hepatic SOD, GSHPX activity and MDA GSH content of

D-gal-induced ageing mice.

Group

Dose

(mg kg�1)

SOD [U (mg

protein)�1]

MDA [nmol

(mg protein)�1]

GSHPX [U (mg

protein)�1)]

GSH [mg

(g protein)�1]

Normal * 82.3793.79** 0.5690.20** 231.27915.52** 65.3696.37**

D-gal * 56.8393.02 1.8990.43 169.33916.71 40.9495.80

MT-a-

glucan

150 66.3092.74** 0.8490.19** 181.28914.08 51.0997.33**

450 72.9494.05** 0.7590.23** 190.73915.09* 56.4296.15**

VE 50 69.6193.66** 0.6990.26** 196.44916.03** 59.5396.93**

Note: Data are the mean9 s.d., n�10. In each vertical column, *P B0.05 and **P B0.01, comparedwith the D-gal group (analysis of variance followed by the Student�Newman�Keuls test).

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The changes in the levels of SOD, GSHpx, GSH and MDA in the liver of normal

mice, D-gal-induced ageing mice and experimental groups are shown in Table 2.

Results showed that the activity of SOD, GSHpx and GSH content in the livers

of D-gal model mice markedly decreased compared with the normal control

(P B0.01). Treatment with MT-a-glucan (450, 150 mg kg�1) and VE (50 mg

kg�1) markedly increased SOD, GSHpx activity and GSH content. MDA content

was markedly increased in D-gal model mice compared with the normal control

(P B0.01). MT-a-glucan (450, 150 mg kg�1) and VE (50 mg kg�1) markedly

decreased MDA. This suggested that MT-a-glucan has an antioxidative effect on

D-gal-induced ageing mice.

Effect of MT-a-glucan on proliferative response and IL-2 production of splenocytesinduced by ConA in D-gal-induced ageing mice

Proliferation and activation of whole spleen cells and of macrophages, combined

with immunocytokines secreted from these immunocytes, were investigated in this

study. IL-2 is a kind of the major and well-known protective immunocytokines

secreted from splenocytes. IL-2 has critical actions on the immune system and has

extensive immuno-enhancing activity. IL-2 remains the most effective cytokine for T

lymphocyte cell expansion and regulation of many critical functions in T-cell biology.

Moreover, IL-2 can augment B-cell proliferation and increase immunoglobulin

synthesis, boost the cytolytic activity of natural killer (NK) cells, and exert actions on

neutrophils and monocytes (Ruth, Carlos, & Gershwin, 2008). The thymus index,

proliferative response and IL-2 production of splenocytes induced by ConA are

shown in Figure 2 and Table 3. The results showed that the thymus index, levels of

proliferative response and IL-2 production of splenocytes were significantly lower in

D-gal-induced ageing mice as compared with normal control (PB0.01 or PB0.05).

Treatment with MT-a-glucan (450, 150 mg kg�1) increased the thymus index, the

proliferative response and IL-2 production markedly. This suggested that MT-

Figure 2. Effect of MT-a-glucan on spleen proliferative response and IL-2 production of

D-gal-induced ageing mice. Data are the mean9s.d., n�10. In each vertical column,

*P B0.05 and **P B0.01, compared with the D-gal group (analysis of variance followed by

the Student�Newman�Keuls test).

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a-glucan has the effect of improving cellular immunity in D-gal-induced ageing

animal model.

Effect of MT-a-glucan on macrophage proliferative response and IL-1 and NOproduction by macrophage in D-gal-induced ageing mice

Macrophages are considered to be one of the effectors that cause the immunitydisruption by producing IL-1 and NO (Hirotada, Noriko, & Hiroaki, 2000). IL-1 is

primarily produced by cells of the mononuclear phagocytic lineage. The biologic

effects and function of IL-1 involve systemic and local effects that have influence on

immunologic properties, including T-cell activation, lymphocyte activating factor,

increasing antibody production and inducing the synthesis of other cytokines. IL-1

also has profound effects on endothelial, smooth muscle, vascular and myocardial

cells. Because of its many, varied, and multiple biologic effects, IL-1 may play a

significant role in the mediation of a number of inflammatory diseases. Cell deathcan be the physiologic response to macrophage activation and overwhelming

expression of IL-1. Nitric oxide (NO), synthesised and secreted by macrophages, is

a message for a wide variety of physiological functions which are synthesised through

the L-arginine pathway by the enzyme NO synthase (NOS) (H.P. Ji, H.K. Ji, & Won,

2006). NOS has three different isoforms: endothelial constitutive NOS (ecNOS),

neural NOS and inducible NOS (iNOS). However, NO is produced in a large amount

by iNOS. Increased NO generation damages DNA, resulting in destruction of cells.

The levels of macrophage proliferative response and IL-1 and NO production bymacrophage are shown in Figure 3 and Table 3.

The levels of macrophage proliferative response and IL-1 and NO production,

and the activity of NOS and iNOS were significantly higher in D-gal model mice as

compared with normal control (PB0.01). Treatment with MT-a-glucan (450, 150 mg

kg�1) decreased the macrophage proliferative response and IL-1 and NO produc-

tion, NOS and iNOS activities markedly. This suggested that MT-a-glucan has the

effect of suppressing production of destructive immune factors in D-gal-induced

ageing animal model. MT-a-glucan could adjust the immunity, which has the effectsof promoting beneficial immune reaction and inhibiting destructive immune

response, and consequently exerting immunomodulatory effect.

In conclusion, MT-a-glucan had an anti-ageing effect on D-gal-treated mice,

which was related to its antioxidative and immunomodulatory effects. Our work can

Table 3. Effect of MT-a-glucan on thymus index and NO production by macrophages in

D-gal-induced-ageing mice.

Group

Dose

(mg kg�1)

Thymus index

mg g�1 BW NO (mmol/L) NOS (U/mL) iNOS (U/mL)

Normal * 1.2290.29** 286.33967.27** 5.4790.24** 3.1690.22**

D-gal * 0.7990.26 504.13970.91 8.5190.62 4.3790.30

MT-a-glucan 150 1.0390.18* 358.06965.70** 6.4990.30** 3.6590.34**

450 1.1290.23* 305.34969.14** 6.7190.55** 3.4090.22**

VE 50 1.0090.19 493.48972.32 7.8290.90 3.9590.47*

Note: Data are the mean9 s.d., n�10. In each vertical column, *P B0.05 and **P B0.01, comparedwith the D-gal group (analysis of variance followed by the Student�Newman�Keuls test).

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be further developed for a healthy food having the effect of delaying senescence,

antioxidation or immunomodulation, which has potential industrial or consumer

application. Further mechanism of action concerning antioxidation and immuno-

modulation will be done in the future study.

Acknowledgements

We acknowledge the funding of this research by the National Natural Science Foundation ofChina (Grant No.31101265).

References

Andras, H., Gyongyi, B., Laszlo, O., Tamas, B., & Gyorgy, K. (2006). Comparison of thecytotoxic effects of receptor tyrosine kinase inhibitors on macrophage functions: Possibleside effects in the immune defense. Immunology Letters, 107, 169�175.

Duan, Z.F., Wang, D.S., & Shen, Y.S. (1993). Study on IL-2 and IL-2 receptor of the aged.Chinese Journal of Geriatrics, 12, 165�169.

Hirotada, K., Noriko, K., & Hiroaki, N. (2000). Activities of polysaccharides obtained fromGrifola frondosa on insulin-dependent diabetes mellitus induced by streptozotocin in mice.Mycoscience, 41, 473�480.

Inoue, A., Kodoma, N., & Nanba, N. (2002). Effect of Maitake (Grifola frondosa) D-fractionon the control of the T lymph node Th-1/Th-2 proportion. Chemical & PharmaceuticalBulletin, 25, 536�540.

Ji, H.P., Ji, H.K., & Won, H.K. (2006). Association of the endothelial nitric oxidesynthase(ecNOS) gene polymorphism with carotidatherosclerosis in type 2 diabetes.Diabetes Research and Clinical Practice, 72, 322�327.

Kubo, K. (1994). Anti-diabetic activity present in the fruit body of Grifola frondosa (maitake).Biological and Pharmaceutical Bulletin, 17, 1106�1110.

Kubo, K. (1997). Anti-hyperliposis effect of maitake fruit body (Grifola frondosa). Biologicaland Pharmaceutical Bulletin, 20, 781�785.

Kubo, K., & Nanba, H. (1998). Modification of cellular immune responses in experimentalautoimmune hepatitis in mice by Maitake (Grifola frondosa). Mycoscience, 39, 351�360.

Lee, B.C., Bae, J.T., Pyo, H.B., Choe, T.B., Kim, S.W., Hwang, H.J., et al. (2003). Biologicalactivities of the polysaccharides produced from submerged culture of the edibleBasidiomycete Grifola frondosa. Enzyme and Microbial Technology, 32, 574�581.

Figure 3. Effect of MT-a-glucan on macrophage proliferative response and IL-1 production

of D-gal-induced-ageing mice. Data are the mean9s.d., n�10. In each vertical column,

*P B0.05 and **P B0.01, compared with the D-gal group (analysis of variance followed by

the Student�Newman�Keuls test).

Food and Agricultural Immunology 417

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ry]

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201

4

Lei, H., Ma, X., & Wu, W.T. (2007). Antidiabetic effect of a (-glucan from fruit body ofmaitake (Grifola frondosa) on KK-Ay mice. Journal of Pharmacy and Pharmacology, 59,575�582.

Lei, H., Wang, B., Li, W.P., Yang, Y., Zhou, A.W., & Chen, M.Z. (2003). Anti-aging effect ofastragalosides and its mechanism of action. Acta Pharmacologica Sinica, 24, 230�234.

Li, X.D., Rong, J.H., & Wu, M.C. (2003). Inhibitory effect of maitake polysaccharide PGF-1on hydroxyl radicals in vitro. Food Science, 24, 126�130.

Liu, X.W., Chen, X.D., & Wu, W.T. (2005). Separation, purification and characterization ofpolysaccharide from Grifola frondosa. Pharmaceutical Biotechnology, 3, 175�178.

Miller, R.A. (1991). Aging and immune function. Cell Immunology, 124, 187�195.Nanba, H., Kodama, N., Schar, D., & Turner, D. (2000). Effects of Maitake(Grifola frondosa)

glucan in HIV-infected patients. Mycoscience, 41, 293�295.Nohl, H. (1993). Involvement of free radicals in aging: a consequence or cause of senescence.

British Medical Bulletin, 46, 653�667.Ooi, V. (1996). Hepatoprotective effect of some edible mushroom. Phytotherapy Research, 6,

536�538.Ruth, Y.L., Carlos, S., & Gershwin, M.E. (2008). The regulatory, inflammatory, and T cell

programming roles of interleukin-2 (IL-2). Journal of Autoimmunity, 31, 7�12.Song, I.K., Kap, S.K., Seok, J.S., Myung, S.K., Dae, Y.K., Sun, L.K., et al. (2007). Anti-

inflammatory effect of Ulmus davidiana Planch (Ulmaceae) on collagen-induced inflam-mation in rats. Environmental Toxicology and Pharmacology, 23, 102�110.

Wang, S.K., Sun, G.J., Zhang, J.X., Jiang, S.K., & Zhang, X.Q. (2002). Development andestimation of an animal subacute aging model induced by D-galactose. Journal of SoutheastUniversity (Medical Science Edition), 21, 217�219.

Zhan, H., Liu, C.G., & Zhou, J.H. (1990). The senile changes of male rats. Acta PhysiologicaSinica, 42, 503�508.

Zhang, X., & Li, W.B. (1990). The biochemical changes of D-galactose treated rats. ChineseJournal of Pharmacology Toxicology, 4, 309�310.

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