Pharmaceutical Biology

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Hypoglycemic Activity of Aqueous Extract of

Pleurotus pulmonarius. in Alloxan-Induced Diabetic
Mice

Sachin L. Badole, Shweta N. Shah, Naimesh M. Patel, Prasad A. Thakurdesai

& Subhash L. Bodhankar

To cite this article: Sachin L. Badole, Shweta N. Shah, Naimesh M. Patel, Prasad A. Thakurdesai
& Subhash L. Bodhankar (2006) Hypoglycemic Activity of Aqueous Extract of Pleurotus
pulmonarius. in Alloxan-Induced Diabetic Mice, Pharmaceutical Biology, 44:6, 421-425, DOI:
10.1080/13880200600794196

To link to this article: https://doi.org/10.1080/13880200600794196

Published online: 07 Oct 2008.

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Pharmaceutical Biology
2006, Vol. 44, No. 6, pp. 421–425

Hypoglycemic Activity of Aqueous Extract of

Pleurotus pulmonarius in Alloxan-Induced Diabetic Mice

Sachin L. Badole, Shweta N. Shah, Naimesh M. Patel, Prasad A. Thakurdesai, and Subhash L. Bodhankar

Department of Pharmacology, Poona College of Pharmacy, Bharati Vidyapeeth Deemed University, Erandwane,
Pune, India

Abstract
Mushrooms have been valued throughout the world as
both food and medicine for thousands of years. In the
current study, we report the hypoglycemic effects of
aqueous extract of one variety of mushroom, Pleurotus
pulmonarius (Fr.) Quel.-Champ (Lentinaceae), in
alloxan-induced diabetic mice. Pleurotus pulmonarius
extract was administrated orally at doses of 250, 500,
and 1000 mg=kg to separate groups of mice (normal
and alloxan-treated mice), and serum glucose and body
weight were measured. In the separate group of mice,
an oral glucose tolerance test (OGTT) was carried out.
Acute oral toxicity data showed no mortality in the nor-
mal mice up to 5000 mg=kg. Oral administration of
extracts reduced the serum glucose level in alloxan-
treated diabetic mice in all the doses tested after acute
and chronic (28 days) administration. The extract also
showed increased glucose tolerance in both normal and
diabetic mice. These results suggest that the extract pos-
sesses hypoglycemic activity.
Keywords: Alloxan-induced diabetic mice, antidiabetic
activity, oral glucose tolerance test, Pleurotus pulmonarius.
Introduction
Mushrooms are a group of fleshy macroscopic fungi that
until recently, as other fungi, were included in the plant
kingdom because of cell wall and spores. Mushrooms
have been valued throughout the world as both food
and medicine for thousands of years (Tribe & Tosco,
1973; Wright, 2004). There are many varieties of mush-
rooms of which Pleurotus are characterized by a white
spore print, attached to gills, often with an eccentric stip,
or no stip at all. They are commonly known as ‘‘oyster
mushrooms’’ (Miles & Chang, 1997). Earlier studies have
reported insulin release and insulin-like activity of other
species of mushroom like Agaricus campestris (Gray &
Flatt, 1998). There is paucity of reports on the hypogly-
cemic activity of Pleurotus. The objective of the current
study was to evaluate the hypoglycemic effect of an aque-
ous extract of Pleurotus pulmonarius (Fr.) Quel.-Champ
(Lentinaceae) on serum glucose in alloxan-induced
diabetic mice.
Materials and Methods
Drugs and chemicals
The mushroom Pleurotus pulmonarius (Fr.) Quel.-
Champ (Lentinaceae) was provided to us as a gift sample
from Bajaj Orchard, Pvt. Ltd. (Mumbai, India). A.M.
Mujumdar (Department of Botany, Agharkar Research
Institute, Pune) authenticated it, and a voucher specimen
was deposited at that institute. Glyburide (Ranbaxy
Pharma. Ltd., New Delhi, India), alloxan monohydrate
(Spectrochem, India), glucose estimation kit (Accurex
Biomedical Pvt. Ltd., Mumbai, India), and D-glucose
(S.D. Fine-Chem. Ltd., Mumbai, India) were purchased
from respective companies.
Experimental animals
Swiss albino mice (25–30 g) were purchased from the
National Toxicology Centre (Pune, India) and used for
the study. They were maintained at a temperature of

Accepted: March 30, 2006

Address correspondence to: Dr. S.L. Bodhankar, Professor and Head, Department of Pharmacology, Bharati Vidyapeeth Deemed
University, Poona College of Pharmacy, Erandwane 411 038, Pune, India. Tel.: þ 91-20-24537237 (ext. 29); Fax: þ 91-20-25439386;
E-mail: sbodh@yahoo.com

DOI: 10.1080/13880200600794196 # 2006 Informa Healthcare

422 S.L. Badole et al.
25
1C and relative humidity of 45% to 55% under 12-h
light:12-h dark cycle. The animals had free access to food
pellets (Chakan Oil Mills, Pune, India), and water was
given ad libitum. The experimental protocol was
approved by the Institutional Animal Ethics Committee
(IAEC) of Poona College of Pharmacy, Pune, India.
Acute oral toxicity studies
Healthy adult albino mice of either sex, starved over-
night, were subjected to acute toxicity studies as per
guidelines (AOT no. 425) suggested by the Organization
for Economic Cooperation and Development (OECD)
(2001). The rats were observed continuously for 2 h for
behavioral, neurological, and autonomic profiles for
any lethality or death for the next 48 h.
Preparation of aqueous extract of Pleurotus pulmonarius
Weighed quantity powder of air-dried Pleurotus pulmonarius
was added to distilled water (1:15), boiled for 20 min on
water bath, cooled to room temperature, and filtered.
The filtrate was dried on a tray dryer at 70C (yield,
24% w=w).The dry extract powder was dissolved in dis-
tilled water to prepare the drug solution of concentration
of 100 mg=ml and used for pharmacological studies.
Induction of experimental diabetes
Diabetes was induced in mice by a single intravenous injec-
tion of aqueous alloxan monohydrate (70 mg=kg, i.v.) by
the method described by Rao et al. (1999). After 48 h,
the animals showing serum glucose levels above 200 mg=dl
(diabetic) were selected for the study. All the animals were
allowed free access to tap water and pellet diet.
Collection of blood and determination of serum glucose
Blood samples from the control and experimental mice
were collected by orbital sinus puncture using hepari-
nized capillary glass tubes. The blood samples so col-
lected were analyzed for glucose levels by the glucose
oxidase peroxidase (GOD=POD) method as described
earlier (Abdel-Barry et al., 1997), and serum glucose
levels were expressed in mg=dl.
Effect of aqueous extract of Pleurotus pulmonarius on
serum glucose in alloxan-induced diabetic mice
The method described by Dunn and McLetchie (1943)
was adopted. Diabetic Swiss albino mice of either sex
were divided into five groups (n ¼ 6), viz.: group I,
vehicle (distilled water, 10 ml=kg); group II, glyburide
(10 mg=kg); group III, extract (250 mg=kg); group IV,
extract (500 mg=kg); group V, extract (1000 mg=kg). All
drugs were given orally.
The acute study involved estimation of serum glucose
at 0, 2, 4, 6, and 24 h after drug administration.
The subacute study involved repeated administration
of drug for 28 days at prefixed times, and serum glucose
levels were estimated on the 7th, 14th, 21st, and 28th
days. The data were represented as mean serum glucose
level and standard error of mean (SEM). The mice were
weighed daily during the study period of 28 days, and
their body weights were noted and presented as mean
change in body weights. The death of mice was also
noted during the study periods, and percentage mortality
was calculated.
Effect of aqueous extract of Pleurotus pulmonarius
on oral glucose tolerance test (OGTT) in normal and
diabetic mice
Normal (nondiabetic) and diabetic mice were divided into
five groups (n ¼ 6), viz.: group I, glucose (2.5 g=kg);
group II, glyburide (10 mg=kg); group III, extract
(250 mg=kg); group IV, extract (500 mg=kg); group V,
extract (1000 mg=kg). The animals were fasted overnight
before commencing the experiment. The animals were
loaded with D-glucose (2.5 g=kg) solution after half an
hour of drug administration. Serum glucose levels were
estimated prior to drug administration and at 30, 60,
and 120 min after glucose loading.
Statistical analysis
The results are expressed as mean
SEM. Comparison
between the groups was made by two-way analysis of
variance (ANOVA) followed by post hoc Dunnett’s test.
Results
Acute toxicity studies revealed that the extract was safe up
to a dose level of 5000 mg=kg of body weight. No lethality
or any toxic reactions were found up to the end of the
study period. Single administration of aqueous extract
of Pleurotus pulmonarius (250, 500, and 1000 mg=kg) as
well as glyburide (10 mg=kg) reduced serum glucose levels
at 2, 4, and 6 h after extract administration (Table 1).
Maximum reduction in serum glucose level was seen at
6 h after extract administration. Subacute administration
(once a day for 28 days) of the extract as well as glyburide
causes significant (p < 0.001) reduction in the serum glu-
cose as compared with vehicle-treated group. Maximum
activity of extract (reduction from 444.29 to 174.32 mg=dl)
was seen with a significant decrease (p < 0.001) in serum
glucose levels at the dose of 500 mg=kg on the 28th day
(Table 2).
Administration of vehicle (distilled water, 10 ml=kg,
p.o.) in alloxan-induced diabetic mice resulted in a
decrease in the body weight during the period of 28 days
(Table 3). Extract (250, 500, and 1000 mg=kg) prevented
 Hypoglycemic activity of Pleurotus pulmonarius 423

Table 1. Effect of Pleurotus pulmonarius on serum glucose level in alloxan-induced diabetic mice (acute study).

Mean fasting glucose level (mg=dl)
SEM

Treatment (mg=kg, p.o.) 0h 2h 4h 6h 24 h

Vehicle 431.79
10.97 434.70
10.80 443.04
13.16 452.43
13.83 456.09
14.67
Glyburide (10) 439.84
19.87ns 358.21
16.14 319.00
15.54 258.86
12.10 421.71
34.27ns
Extract (250) 456.42
26.14ns 327.16
27.33 292.78
22.44 256.18
23.92 344.07
31.19
Extract (500) 444.29
13.05ns 148.22
15.24 131.24
9.59 121.10
13.55 345.09
9.55
Extract (1000) 444.92
30.39ns 286.90
16.50 230.70
15.21 160.59
14.22 222.25
25.64

n ¼ 6, data were analyzed by two-way ANOVA followed by post hoc Dunnett’s test. ns, not significant.

p < 0.05 compared with vehicle treated group (distilled water, 10 ml=kg). All other values are significant (p < 0.001) as compared
with vehicle-treated group.

Table 2. Effect of Pleurotus pulmonarius on serum glucose level in alloxan-induced diabetic mice (subacute study).

Mean fasting glucose level (mg=dl)
SEM

Treatment (mg=kg, p.o.) Day 0 Day 7 Day 14 Day 21 Day 28

Vehicle 431.79
10.97 474.38
6.57 479.23
4.88 493.95
11.98 503.26
15.56
Glyburide (10) 469.84
19.87ns 367.54
14.51 297.61
17.55 264.85
14.83 196.25
18.45
Extract (250) 456.42
26.14ns 312.37
19.74 241.68
9.51 214.27
9.92 204.98
9.93
Extract (500) 444.29
13.05ns 261.87
14.64 190.33
15.46 182.78
18.45 174.32
14.83
Extract (1000) 444.92
30.39ns 314.39
15.95 275.41
21.79 240.24
15.56 185.56
11.98

n ¼ 6, data were analyzed by two-way ANOVA followed by post hoc Dunnett’s test. ns, not significant.
All the values are significant (p < 0.001) as compared with vehicle-treated group (distilled water, 10 ml=kg).

a decrease in body weight in alloxan-treated mice. On the
other hand, mice gained body weight as compared with
the vehicle-treated group, which indicated the beneficial
effect of the extract in preventing further loss of body
weight. Administration of vehicle in alloxan-induced dia-
betic mice resulted in death of 44.4% of the total animals
during the 28-day study period. Administration of the
extract (250, 500, and 1000 mg=kg) reduced mortality
(14.29%, 14.29%, and 25%, respectively) in mice. It
was thus apparent that when no drug was administered,
progression of diabetes resulted in mortality of mice,
whereas the extract treatment resulted in reduction of
mortality. Extract (500 mg=kg) produced a significant
(p < 0.05) increase in the glucose threshold, 30-min
post–glucose loading in both normal (Table 4) as well
as diabetic mice (Table 5).
Discussion
Mushrooms are highly nutritive as they contain good-
quality proteins, vitamins, and minerals (Flegg & Maw,
1976; Khanna & Garcha, 1984). Mushrooms are a low-
calorie food with very little fat and are highly suitable
for obese persons. With no starch and very low sugars,
they are the ‘‘delight of the diabetics’’ (Bano, 1976). In

Table 3. Effect of extract on body weight in alloxan-induced diabetic mice.

Mean body weight (g)
SEM

Treatment (mg=kg, p.o.) 0h Day 7 Day 14 Day 21 Day 28

Vehicle 32.00
1.41 26.33
0.75 24.33
0.66 22.33
0.66 17.83
0.60
Glyburide (10) 36.17
1.33ns 34.37
0.36 36.00
2.39 36.33
2.38 37.00
1.20
Extract (250) 33.17
2.04ns 32.83
1.44 31.00
1.2 30.33
1.20 29.50
0.99
Extract (500) 31.17
2.04ns 32.00
0.6 33.00
0.70 33.00
0.70 33.10
0.56
Extract (1000) 35.50
1.64ns 33.00
0.63 32.00
0.49 30.67
0.49 29.67
1.23

n ¼ 6, data were analyzed by two-way ANOVA followed by post hoc Dunnett’s test. ns, not significant.

p < 0.05 compared with vehicle-treated group (distilled water, 10 ml=kg). All other values are not significant as compared with
vehicle-treated group.
424 S.L. Badole et al.

Table 4. Effect of Pleurotus pulmonarius on oral glucose tolerance test (OGTT) in normal mice.

Mean fasting glucose level (mg=dl)
SEM

Treatment (mg=kg, p.o.) Before glucose 0 min 30 min 60 min 120 min

Vehicle 120.48
14.26 318.03
15.13 197.42
20.24 142.83
121.69 136.58
15.64
Glyburide (10) 117.44
14.04 289.83
20.71 203.78
28.00 121.69
21.38 169.34
29.38
Extract (250) 106.05
5.34 328.92
12.50 252.98
12.35 172.31
13.65 143.58
7.28
Extract (500) 112.01
8.42 319.55
11.88 121.74
11.32 126.10
17.96 138.02
18.34
Extract (1000) 105.51
15.17 288.04
12.77 190.14
23.11 180.12
15.16 132.08
11.42

n ¼ 6, data were analyzed by two-way ANOVA followed by post hoc Dunnett’s test.

p < 0.05,  p < 0.01,  p < 0.001, compared with vehicle-treated group (distilled water, 10 ml=kg).

adequate quantities and being low in sugars, they can
serve as medicinal foods for diabetes (Rai, 1986). Pre-
liminary phytochemical analysis of the Pleurotus pulmo-
narius showed the presence of proteins, minerals,
vitamins, and carbohydrates (Food and Agriculture
Organization of the United Nations, 1968).
In the past, many mushroom varieties have been
reported to possess hypoglycemic activities in animals
(Gray & Flatt, 1998; Swanston-Flatt et al., 1989), as well
as in diabetic patients (Konno et al., 2001). In normal and
streptozotocin diabetic mice treated with the mushroom
variety Agaricus bisporus (J. Lange) Imbach, the mush-
room was shown to retard the development of hypergly-
cemia, hyperphagia, polydipsia, body weight loss, and
glycated hemoglobin in the streptozotocin-treated mice
(Swanston-Flatt et al., 1989) by counteracting reduction
in plasma and pancreatic insulin concentration and by
improving the hypoglycemic effect of exogenous insulin.
In the current study, the hypoglycemic activity of the
aqueous extract of Pleurotus pulmonarius was evaluated
in alloxan-induced diabetic mice. Significant reduction
in serum glucose level was seen at the second hour and
maximum reduction occurred at the sixth hour by treat-
ment with the extract in an acute study, and also on 28-
day administration. The extract showed short onset and
prolonged duration of hypoglycemic action. A dose of
500 mg=kg of extract showed optimum activity, as com-
pared with 250 and 1000 mg=kg doses of extract.
Subacute treatment for 28 days with the extract in the
tested doses brought about improvement in body weights
of alloxan-treated diabetic mice, indicating its beneficial
effect in preventing loss of body weight in diabetic mice.
Administration of extract lowered mortality (14.29%) as
compared with alloxan-induced diabetic mice (44%).
The protective effects against diabetes-induced weight
loss is supported by earlier studies (Swanston-Flatt
et al., 1989).
In the oral glucose tolerance test, the doses increased
the tolerance for glucose suggesting increased peripheral
utilization of glucose in both diabetic as well as non-
diabetic (normal) mice. The extract showed optimum
activity at the dose of 500 mg=kg. Agaricus campestris
L. Fr., a mushroom variety, was reported to counter the
hyperglycemia of streptozotocin-diabetic mice probably
by an insulin-releasing mechanism (Gray & Flatt, 1998).
Water-soluble fraction obtained from maitake mush-
room is reported to lower fasting blood glucose and, thus,
is useful to treat insulin resistance in animals (Talpur
et al., 2002a, 2002b), as well as in diabetic patients
(Konno et al., 2001). This effect of maitake mushroom
was suggested to be through glucose=insulin metabolism
and=or by enhancing peripheral insulin sensitivity. A
large amount of glycogen was also observed after treating
rats with maitake mushrooms, suggesting the possibility
of increased glycogen formation by mushroom as a prob-
able mechanism of their hypoglycemic effect.

Table 5. Effect of Pleurotus pulmonarius on oral glucose tolerance test (OGTT) in diabetic mice.

Mean fasting glucose level (mg=dl)
SEM

Treatment (mg=kg, p.o.) Before glucose 0 min 30 min 60 min 120 min

Vehicle 421.53
18.59 511.76
12.47 457.46
11.22 335.902
11.02 534.62
12.16
Glyburide (10) 474.80
20.85 514.38
6.47 333.05
5.13 339.13
9.56 472.68
27.28
Extract (250) 491.62
25.55 528.73
13.50 477.05
7.13 454.67
7.12 489.52
13.73
Extract (500) 489.35
20.75 542.69
8.44 310.43
8.37 326.69
10.29 485.56
10.39
Extract (1000) 480.16
19.45 546.66
13.56 476.35
8.92 454.67
10.63 503.52
17.44

n ¼ 6, data were analyzed by two-way ANOVA followed by post hoc Dunnett’s test.

p < 0.05,  p < 0.001, compared with vehicle-treated group (distilled water, 10 ml=kg).
 Hypoglycemic activity of Pleurotus pulmonarius
The ability of lectins isolated from mushrooms (A.
campestris, A. bisporus) to enhance insulin release by iso-
lated rat islets of Langerhans has been documented
(Ewart et al., 1975). The nature of the active principle(s)
and mechanism of action of insulin secreting cells and
muscles remain to be established (Gray & Flatt, 1998).
Some variety of mushrooms were also shown to pos-
sess antihypertensive effects in spontaneous hypertensive
rats (Talpur et al., 2002b), indicating the possibility of
antidiabetic potential of mushrooms for hypertensive
patients.
Conclusions
In conclusion, an oral dose of 250 and 500 mg=kg of aque-
ous extract of Pleurotus pulmonarius possesses strong
hypoglycemic activity against alloxan-induced diabetes
and increased oral glucose tolerance in diabetic (OGTT
model) mice. An upper dose (1000 mg=kg) caused an
increase in mortality and loss of body weight without
an increase in the hypoglycemic effect in diabetic mice.
Acknowledgments
The authors would like to acknowledge Dr. S.S. Kadam,
Principal, and Dr. K.R. Mahadik, Vice-Principal, Poona
College of Pharmacy, Bharati Vidyapeeth Deemed Uni-
versity, Pune, for providing necessary facilities to carry
out the study. We are also thankful to Bajaj Orchard, a
division of Trinity Bio-Tech, Mumbai, India, for a gift
sample of Pleurotus pulmonarius.
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