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Renu Anna Joseph. et al/ International Journal of Pharmaceutical Research and life Sciences. 1(2), 2013, 174 - 179.

ISSN: 2321 4589

International Journal of
Pharmaceutical Research and
Life Sciences

IJPRLS

Journal home page: www.ijprls.com

STUDY ON THE GLUCOSE ADSORPTION CAPACITY AND EFFECT ON GLUCOSE UPTAKE BY


YEAST CELLS OF AQUEOUS EXTRACTS OF TRIGONELLA FOENUM-GRACEUM SEEDS&
BOERHAAVIA DIFFUSA LEAVES.
Renu Anna Joseph*, Saana Sain, Christa. J. George, Vipin Varghese, Sujith. K
Pushpagiri College of Pharmacy, Tiruvalla, Kerala.
ABSTRACT
The present study evaluated the hypoglycemic activity of aqueous extracts of trigonella foenum-graceum
seeds& boerhaavia diffusa leaves by means of invitro methods. The evaluation was done by studying their
effect on glucose adsorption and glucose uptake by yeast cells. The study has revealed that all the taken
samples adsorbed glucose to an extent and the enhancement of glucose uptake was dependent on both the
samples and glucose concentration.
Keywords: Hypoglycaemic, Glucose concentration, Evaluation.

Author for Correspondence:


Renu Anna Joseph
Pushpagiri College of pharmacy, Tiruvalla,
kerala.
E mail: sujipillai76@yahoo.com

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INTRODUCTION
In recent years, evidence of cases of insulin
resistance and the occurrence of side effects from
prolonged administration of conventional drugs have
triggered the search for safe and effective
alternatives. Nowadays, medicinal plants play an
important role in the treatment of diabetes mellitus.
Several plant extracts and isolated phytochemical
have been examined for the antidiabetic activity,
with a view to identify alternative treatment
strategies for diabetes. It has been observed that,
certain resistant cases of diabetes, that do not
respond well to conventional drugs often respond
well
to
supplementation
with
natural
1
remedies .Seeds of Trigonella foenum graceum
(fenugreek) commonly known as methi belongs to
the family Leguminosae. It is used as food and also
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Renu Anna Joseph. et al/ International Journal of Pharmaceutical Research and life Sciences. 1(2), 2013, 174 - 179.

for the medicinal purposes. It is good source of


many essential elements such as iron, phosphorous
and sulphur. Seeds can inhibit cancer of the liver,
lower blood cholesterol levels and also have an
antidiabetic effect2,3. Leaves of Boerhaavia diffusa
commonly known as punarnava, belongs to the
family Nyctaginaceae. The extracts are also useful in
diabetic patients, in the prevention and treatment of
diabetes
induced
hyperlipidemia
and
4
atherosclerosis . But the scientific validation for
their precise mode of action has not yet been
reported.With this background, aqueous extracts of
seeds of Trigonella foenum graceum and leaves of
Boerhaavia diffusa were selected, for which no
reports are available on the mechanisms of their
hypoglycemic effect. Therefore, the present study
was aimed to determine the mechanisms, through
which they produce their hypoglycemic effect.
MATERIALS AND METHODS
Preparation of aqueous extract
Aqueous extracts were prepared by percolation
process where the powdered materials were
uniformly moistened with water for a period of four
hours in a separable vessel and were packed evenly
into a percolator. Sufficient water is poured over the
drug and allowed to stand for 24 hours. Percolate
was collected and the marc was pressed and the
expressed liquid is added to the percolate giving 8090% of final volume. Evaporated and concentrated
to get the finished products. The product is then
redissolved in water to get the final concentration of
10mg/mL5.
Effect of extracts on adsorption of glucose
Glucose adsorption capacity of the samples was
determined according to the method of Ou et al6.
One gram of the sample was added to 100 ml of
glucose solution of increasing concentration
(5mmol/L, 15mmol/L, 25mmol/L, 50mmol/L, and
100mmol/L). The mixture was stirred well,
incubated in shaker water bath at 370C for at 6
hours, centrifuged at 4000g for 20 minutes7. The
glucose content in the supernatant was determined.
Glucose bound was calculated using formula:

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Glucose bound (mM) =

G1 G6 Volume of solution (ml)


Weight of the sample (mg)

G1: glucose concentration of original solution


(mg/dl)
G6: glucose concentration after 6 hours (mg/dl)
Effect extracts on glucose uptake by yeast cells
Preparation of yeast cells
Yeast cells were prepared according to the method
of Cirillo 8. Commercial bakers yeast was washed
by repeated centrifugation (3000g, 5 minutes) in
distilled water until the supernatant fluids were clear
and a 10% (v/v) suspension was prepared in distilled
water9.
Glucose uptake by yeast cells
Various concentrations of the aqueous extracts
(1-5mg) were added to the 1 ml of glucose solution
(5, 15, 25, 50,100mM) and incubated together for 10
minutes at 370C. Reaction was started by adding 100
l of yeast suspension, vortex and further incubated
at 370C for 60 minutes. After 60 minutes the tubes
were centrifuged (2500g, 5 minutes) and glucose
was estimated in the supernatant10,11. The
experiments were carried out in triplicate .The
percent increase in the glucose uptake was
calculated using the formula:
Percentage increasein glucose uptake = Absorbance of control
Absorbance of sample/ Absorbance of control 100

RESULTS
The glucose adsorption capacity of aqueous extracts
of trigonella foenum-graceum seeds& boerhaavia
diffusa are shown in table No.1. A series of different
concentrations of glucose (5-100mM) were used for
determining the adsorption capacity of samples for
glucose. The results have showed that aqueous
extracts of seeds of Trigonella foenum graceum
showed significantly higher glucose adsorption
capacity at 100mM when compared to Boerhaavia
Diffusa. The effect of aqueous extracts of trigonella
foenum-graceum seeds& boerhaavia diffusa on
glucose transport across yeast cell membrane was
studied in an invitro system comprising of yeast cells
suspended in glucose solution of varying
concentration (5-100mM) in presence of extracts.
The percentage increase in glucose uptake by yeast
cells is presented in Table No. 2,3,4,5 & 6.The

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Renu Anna Joseph. et al/ International Journal of Pharmaceutical Research and life Sciences. 1(2), 2013, 174 - 179.

amount of glucose remaining in the medium after a


specific time serves as an indicator of the glucose
uptake by the yeast cells. The plant extracts showed
an increase in glucose uptake by yeast cells.
However, the increase in glucose uptake produced
by various concentrations of Trigonella foenum
graceum seed extract and Boerhaavia diffusa leaf

extract in different molar concentration of glucose


were comparable. The increase in glucose uptake
was dose dependant and it was directly proportional
to the sample concentration. However, the percent
increase in glucose uptake by the yeast cell was
inversely proportional to the glucose concentration.

Table No.1: Glucose Bound by Medicinal Plant Extracts in Different Concentrations of Glucose
GLUCOSE BOUND (mM)
SAMPLE

5mM

15mM

25mM

50mM

100mM

Trigonella foenum graceum

3.9210.6

12.2100.7

13.5121.7

38.4152.3

79.9213.5

Boerhaavia diffusa

1.4760.3

13.2450.5

14.9950.9

29.9902.4

60.9802.4

Values are expressed as mean SEM(n=3)


Table No.2: Effect of Aqueous Extracts on Glucose Uptake by Yeast Cells in Different Concentrations
of Glucose
% INCREASE IN GLUCOSE UPTAKE
SAMPLE

1mg/ml

2mg/ml

3mg/ml

4mg/ml

5mg/ml

Trigonella foenum graceum

80.2280.4

81.4291.3

82.7152.4

83.0101.4

85.7533.2

Boerhaavia diffusa

77.3020.6

78.6891.2

80.9252.7

81.1431.4

84.8542.8

Values are expressed as mean SEM (n=3)


Table No.3: Showing Percentage Increase in Glucose Uptake by Yeast Cells in 15mm of Glucose
% INCREASE IN GLUCOSE UPTAKE
SAMPLE

1mg/ml

2mg/ml

3mg/ml

4mg/ml

5mg/ml

Trigonella foenum graceum

79.4480.5

80.6441.2

81.8232.4

82.7383.0

83.1462.5

Boerhaavia diffusa

74.5490.6

77.9121.3

79.0582.8

80.4371.9

82.6632.8

Values are expressed as mean SEM (n=3)

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Renu Anna Joseph. et al/ International Journal of Pharmaceutical Research and life Sciences. 1(2), 2013, 174 - 179.

Table No.4: Showing Percentage Increase in Glucose Uptake by Yeast Cells in 25mm of Glucose
% INCREASE IN GLUCOSE UPTAKE
SAMPLE

1mg/ml

2mg/ml

3mg/ml

4mg/ml

5mg/ml

Trigonella foenum graceum

77.7321.7

77.8422.0

78.7182.6

79.9042.7

79.9995.5

Boerhaavia diffusa

72.9351.2

75.3142.5

77.1122.6

79.8422.3

79.8425.2

Values are expressed as mean SEM (n=3)


Table No.5: Showing Percentage Increase in Glucose Uptake by Yeast Cells in 50mm of Glucose
% INCREASE IN GLUCOSE UPTAKE
SAMPLE

1mg/ml

2mg/ml

3mg/ml

4mg/ml

5mg/ml

Trigonella foenum graceum

72.1121.5

76.4202.7

77.1204.1

77.8544.8

78.1125.4

Boerhaavia diffusa

70.0101.5

72.5102.9

75.5054.7

78.0104.9

78.4324.8

Values are expressed as mean SEM (n=3)


Table No.6: Showing Percentage Increase in Glucose Uptake by Yeast Cells in 100mm of Glucose
% INCREASE IN GLUCOSE UPTAKE
Trigonella foenum graceum

72.1121.5

76.4201.9

77.1201.6

77.8543.0

78.1123.7

Boerhaavia diffusa

70.0101.8

72.5101.5

75.5053.7

78.0102.9

78.4324.9

Values are expressed as mean SEM (n=3)


DISCUSSION AND CONCLUSION
The present study reports the effect of aqueous
extracts of trigonella foenum-graceum seeds&
boerhaavia diffusa leaves on glucose uptake by
yeast cells and glucose adsorption capacity. The
results revealed that exhibited glucose adsorption
and an increase in the glucose uptake by yeast cell.
The mechanism of glucose transport across the yeast
cell membrane has been receiving attention as an in
vitro screening method for the hypoglycemic activity
of various compounds in medicinal plants12,13. In this
study, it was observed that the increase in glucose
uptake by yeast cells occurred in a dose dependant
manner and the rate of glucose transport across the
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yeast cell membrane was the function of external


glucose concentration as well as sample
concentration. However this relationship seem to
seize, when the yeast cells has reached a saturation
point. The glucose transport in yeast cells is
extremely complex and the sugar transport across
the yeast cell membrane is mediated by facilitated
diffusion process, in which a number of
stereospecific membrane carriers are involved14.
Facilitated carriers are specific carriers that transport
solutes down the concentration gradient. This
suggests that the effective transport of glucose is
only attained, if there is removal of intracellular
glucose15. In this study the extracts produced an
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Renu Anna Joseph. et al/ International Journal of Pharmaceutical Research and life Sciences. 1(2), 2013, 174 - 179.

increase in glucose uptake. The aqueous extracts of


trigonella foenum-graceum seeds& boerhaavia
diffusa leaves exhibited a higher glucose uptake.
However the aqueous extracts of seeds of Trigonella
foenum graceum exhibited higher glucose adsorption
capacity than boerhaavia diffusa leaves. The glucose
adsorption capacities of the aqueous extracts were
directly proportional to the glucose concentration in

the medium resulting in significantly higher glucose


adsorption with an increase in glucose concentration.
The extacts have adsorbed some amount of glucose
even at low concentration of glucose, which shows
that the sample might help to retain the glucose in
the intestinal lumen even at low glucose
concentration.

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Renu Anna Joseph. et al/ International Journal of Pharmaceutical Research and life Sciences. 1(2), 2013, 174 - 179.

14. Neile K. Edens, Lisa A. Reaves, Marti S. Bergana, Irene L. Reyzer, Patrick O. Mara, Jeffrey H. Baxter,
Maureen K.Snowden. Yeast extract stimulates glucose metabolism and inhibits lipolysis in rat adipocytes
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