Indo American Journal of Pharmaceutical Research, 2015
ISSN NO: 2231-6876
IN VITRO GLUCOSE UPTAKE ACTIVITY OF AN OLEANANE-TYPE TRITERPENOID
SAPONIN ISOLATED FROM MOMORDICA CYMBALARIA
Mr. Suman Samaddar1, Dr. Raju Koneri Balwanth2, Mr. Abhinaya Bhattarai2 & Dr. K.B.
Chandrasekhar3
1
Research Scholar, Department of Pharmaceutical Sciences, Jawaharlal Nehru Technological University, Anantapur, Andhra
Pradesh, India.
2
Department of Pharmacology, Karnataka College of Pharmacy, Bangalore, India.
3
Research & Development Cell, Jawaharlal Nehru Technological University, Anantapur, Andhra Pradesh, India.
ARTICLE INFO
Article history
Received 16/05/2015
Available online
30/05/2015
Keywords
Oleanane-Type Triterpenoid
Saponin,
Momordica Cymbalaria,
In Vitro Glucose Uptake
Assay,
GOD-POD Method,
MTT Assay,
Diaphragm.
ABSTRACT
The antidiabetic activity of Momordica cymbalaria was investigated specifically in its ability
to increase the uptake of glucose. A triterpenoid saponin of oleanane-type (SMC) was isolated
from the roots of Momordica cymbalaria. Glucose uptake experiment by isolated diaphragms
of both diabetic, following streptozotocin administration, and non-diabetic animals (Swiss
Albino mice) was carried in the presence and absence of SMC. To determine the in vitro
cytotoxicity, MTT assay was performed on L6 cell line (mouse skeletal muscle) for test dose
fixation. Further, glucose uptake by L6 cell line in the presence of SMC was performed.
Glucose uptake was determined by glucose oxidase-peroxidase (GOD-POD) method. SMC
did not show cytotoxicity against L6 cells and significantly increased glucose uptake up to
242.75% over control in a dose-dependent manner. Diabetic and non-diabetic diaphragms
also showed increase in glucose uptake in the presence of SMC over control. Data from both
the models (cells and isolated diaphragms) indicate that SMC enhances glucose uptake
thereby exhibiting hypoglycemic activity. The findings suggest that SMC can prove to be
very effective against hyperglycemia due to insulin resistance, as in Type 2 Diabetes Mellitus.
Copy right © 2015 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical
Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
www.iajpr.com
Page
Please cite this article in press as Mr. Suman Samaddar et al. In vitro glucose uptake activity of an oleanane-type triterpenoid
saponin isolated from Momordica cymbalaria. Indo American Journal of Pharm Research.2015:5(05).
2071
Corresponding author
Mr. Suman Samaddar
Karnataka College of Pharmacy,
#33/2, Thirumenahalli,
Hegde Nagar Main Road, Bangalore – 560064,
080-65332053,
+91-9972809052,
sumanpppa@gmail.com
Vol 5, Issue 05, 2015.
Mr. Suman Samaddar et al.
ISSN NO: 2231-6876
INTRODUCTION
Diabetes mellitus is becoming the third killer of mankind after cancer and cardiovascular disease due to its high prevalence,
morbidity and mortality [1]. Among the currently available therapeutics, sulfonylureas are exclusively used for stimulation of the βcells to release more insulin. Chronic hyperglycemia is associated with prolonged damage, malfunctioning and eventually failure of
organs, especially the kidney, nerves, heart, eyes and blood vessels [2]. There is a need for antidiabetic drug with multiple target and
potency. The need of herbal remedies has gone up owing to their fewer side effects, efficacy and relatively less treatment costs. Drugs
of natural origin or their extracts are prescribed widely, even when their biologically active components are not known. The World
Health Organization (WHO) even approves the use of natural drugs of plant origin for the treatment of different diseases, including
diabetes mellitus [3, 4].
Skeletal muscle is one of the major insulin-target tissues responsible for maintenance of whole body glucose homeostasis. It
is well established that insulin stimulation of glucose uptake in skeletal muscle cells is mediated through translocation of glucose
transporter 4 (GLUT4) from the endoplasmic reticulum to the plasma membrane [5]. It is also known that a defect in glucose transport
efficiency and GLUT4 activity results in insulin resistance [6]. L6 cells represent a good model for glucose uptake because they have
been used extensively to elucidate the mechanisms of glucose uptake in muscle, have an intact insulin signaling pathway, and express
the insulin-sensitive GLUT4 transporters [7].
Momordica cymbalaria Fenzl (Cucurbitaceae) is a species found in the states of Karnataka and Andhra Pradesh, India. Its
tuber is traditionally used as an abortifacient [8] and the fruit powder and extracts of MC were previously reported to have Type 1
antidiabetic activity in experimental diabetic models [9, 10]. Its various mechanisms of action have been reported in diabetic rats [11].
Antiovulatory, abortifacient, anti-implantation and cardio-protective activities have also been reported [12-14]. The antidiabetic
activity of saponins of Momordica cymbalaria may be due to reversing of the atrophy of the pancreatic islets of β-cells, as a result of
which there may be increased insulin secretion and increase in the hepatic glycogen level and these may attenuate hyperinsulinaemia.
The α-adrenergic blocking effect might contribute to their insulin secretion and sensitizing effects [10]. Accordingly, the objective of
the current study is to determine whether the active phytomolecule – an oleanane-type triterpenoid saponin isolated from the roots of
Momordica cymbalaria [15] has any influence on the uptake of glucose L6 skeletal muscle cell line and isolated diaphragms of
diabetic and non-diabetic mice. Additionally, we also determined the cytotoxicity of SMC on L6 cells.
MATERIALS AND METHODS
Extraction and isolation of saponin
The oleanane-type triterpenoid saponin was extracted from the roots of Momordica cymbalaria [15].
Chemicals
Streptozotocin (Sigma Aldrich), 3-(4,5–dimethyl thiazol–2–yl)–5–diphenyltetrazolium bromide (MTT), Fetal Bovine serum
(FBS), Phosphate Buffered Saline (PBS), Kreb’s-Ringer bicarbonate buffer, Dulbecco’s Modified Eagle’s Medium (DMEM), glycine,
EDTA, Glucose, antibiotics and insulin were obtained from Hi-Media Laboratories Ltd., Mumbai. Dimethyl Sulfoxide (DMSO) and
2-Propanol from Merck Ltd., Mumbai, India, and TrypLE TM Express (cell dissociation reagent) from Invitrogen, Life Technologies,
USA. Metformin hydrochloride was obtained from Sigma-Aldrich, USA. Glucose oxidase-peroxidase (GOD-POD) kit from DiaSys
Diagnostic Systems, Germany.
Animals
Page
Induction of experimental diabetes
Overnight fasted animals were injected with streptozotocin dissolved in 0.1 M citrate buffer (pH 4.5) at a dose of 65 mg/kg.
The animals were allowed to drink 5% w/v glucose solution to overcome the drug-induced hypoglycemia. On the third day animals
with blood glucose level higher than 200 mg/dL were considered as diabetic and were used for further studies [16, 17].
In the acute oral toxicity study, mortality was seen in the main test at 1750 mg/kg. Hence, 175mg/kg was concluded as safe dose. For
the study, 100 mg/kg was selected as the dose. The animals (non-diabetic and diabetic) were anesthetized using anesthetic ether and
the diaphragms were dissected out quickly with minimal trauma. The diaphragms were then rinsed in cold Kreb’s-Ringer bicarbonate
buffer (without glucose) to remove any blood clots and were placed in small culture tubes containing 2 ml Kreb’s-Ringer bicarbonate
buffer with 2% glucose and incubated for 30 min at in an atmosphere of 95% O 2 and 5% CO2 with shaking at 140 cycles per min.
Following incubation, the glucose content of the incubated medium was measured by glucose oxidase-peroxidase (GOD-POD)
method [18]. The uptake of glucose was calculated in mg/g of moist tissue/30 min. Glucose uptake per gram of tissue was calculated
as the difference between the initial and final glucose content in the incubated medium.
2072
Male Swiss albino mice weighing between 25-35 g were maintained in standard laboratory conditions at room temperature
(25±2 0C) with 12:12 h L:D. The animals were given pellet chow and water ad libitum except during experimentation. The study
protocols were duly approved by the Institutional Animal Ethics Committee (IAEC) at Karnataka College of Pharmacy, Bangalore.
Studies were performed in accordance with the CPCSEA guidelines. The oral acute toxicity was performed using the ‘up and down’
procedure of OPPTS (guideline no. 870.1100).
www.iajpr.com
Vol 5, Issue 05, 2015.
Mr. Suman Samaddar et al.
ISSN NO: 2231-6876
Glucose uptake by diaphragm of non-diabetic (healthy) mice
The animals were divided into the following 4 groups containing 6 mice each:
Gr. I: Normal control (2 mL Tyrode solution with 2% glucose)
Gr. II: Normal control + Insulin (0.25 IU/mL)
Gr. III: Normal control + metformin (1 mg/mL)
Gr. IV: Normal control + SMC (1 mg/mL)
Glucose uptake by diaphragm of diabetic mice
The animals were divided into the following 5 groups containing 6 mice each:
Gr. I: normal control (distilled water po once daily)
Gr. II: diabetic control (streptozotocin 65mg/kg ip/single dose)
Gr. III: Diabetic control + Insulin (4 IU/kg/i.p/day/30 days)
Gr. IV: diabetic control + metformin (50 mg/kg/30 days).
Gr. V: diabetic control + SMC (100 mg/kg, po/30 days)
Cell culture
L-6 cell cultures were procured from National Centre for Cell Sciences (NCCS), Pune, India. Stock cells were cultured in
Dulbecco’s modified Eagle’s medium (DMEM). Medium was supplemented with 10% Fetal Bovine Serum (FBS), Penicillin (100
IU/ml), streptomycin (100 μg/ml) and amphotericin B (2.5 μg/ml) in an humidified atmosphere of 5% CO 2 at 37°C until confluent.
The cells were dissociated with TrypLETM Express solution. The stock cultures were grown in T75 culture flasks.
Determination of Cell Viability by MTT Assay
The assessment of cytotoxicity of SMC on L6 cells was done by the reduction of 3-(4,5- dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide (MTT) to formazan. Cells were seeded in a 96-well plate (0.5-10×103 cells/well), and left to attach on to
the substrate overnight before being exposed to SMC, and incubated at 37 oC for 24 h. After 36 h 50 μL MTT (5 mg/mL) solution was
added to each well and the cells were incubated in the dark at 37°C for an additional 4 h. Thereafter, the medium was removed, the
formazan crystals were dissolved in 200 μL of DMSO and 25 μL of glycice buffer (pH 10.5), and the absorbance was measured at 570
nm in a micro plate reader. The percentage cell viability was calculated [19].
Glucose uptake assay
Cells were cultured on 96-well plates and incubated for 48 h at 37°C in CO 2 incubator. When semi-confluent monolayer was
formed the culture was renewed with serum free-DMEM and incubated for 18 h at 37°C. After 18 h the medium was discarded and
cells were washed with PBS once. The cells were treated with insulin, standard drug (metformin), SMC and added glucose (5.5mM)
and incubated for 30 mins. The supernatant was collected for glucose estimation by GOD-POD method [20].
Grouping of cells:
Four groups of cells (n=4) were considered for the assay:
Group 1: Cells exposed to glucose (5.5mM) – serves as Glucose control.
Group 2: Glucose control + Insulin (0.1-1.0 IU)
Group 3: Glucose control + Metformin (0.01-0.1 mg)
Group 4: Glucose control + SMC (0.01-0.1 mg)
RESULTS
Glucose uptake by diaphragm of non-diabetic (healthy) mice
Saponin of M. cymbalaria enhanced the uptake of glucose in mice diaphragm (P <0.001) when compared to control group. It
was also comparable to the glucose uptake in mice diaphragm mediated by metformin (Table 1).
Table 1: Effect of saponin of Momordica cymbalaria on glucose uptake in non-diabetic (healthy) mice diaphragm.
Glucose uptake by diabetic mice diaphragm
SMC treated diabetic mice diaphragm (Gr. IV) showed significant increase (P <0.01) in glucose when compared to diabetic
control mice diaphragm (Gr. II) and the results are similar to metformin-mediated action (Table 2).
www.iajpr.com
2073
Glucose uptake by diaphragm (mg/g diaphragm)
2.03 ± 0.23
4.04 ± 0.47*
7.51 ± 0.21*
6.94 ± 0.21*
Page
Treatment Group
Gr. I: Normal control (2 mL Tyrode solution with 2% glucose)
Gr. II: Normal control + Insulin (0.25 IU/mL)
Gr. III: Normal control + Metformin (1 mg/mL)
Gr. IV: Normal control + SMC (1 mg/mL)
*p>0.001, when compared to control group.
Values are expressed as mean± SEM of 6 animals (n=6) in each group.
Vol 5, Issue 05, 2015.
Mr. Suman Samaddar et al.
ISSN NO: 2231-6876
Table 2: Effect of saponin of Momordica cymbalaria (100 mg/kg, po/30 days) on glucose uptake in STZ-induced (65 mg/kg,
i.p./single dose) diabetic mice diaphragm.
Treatment Group
Gr. I: Normal control (distilled water po once daily)
Gr. II: Diabetic control (65 mg/kg, i.p/single dose)
Gr. III: Diabetic control + Insulin (4 IU/kg/i.p/day/30 days)
Gr. IV: Diabetic control + Metformin (50 mg/kg/30 days)
Gr. V: Diabetic control + SMC (100 mg/kg, po/30 days)
*p>0.01, when compared to # normal control, * diabetic control group.
Values are expressed as mean± SEM of 6 animals (n=6) in each group.
Glucose uptake by diaphragm (mg/g diaphragm)
2.4 ± 0.3
1.1 ± 0.14#
2.19 ± 0.31*
2.21 ± 0.24*
2.28 ± 0.27*
Cytotoxicity of SMC on L6 cells
The percentage viability of L6 cells after treatment with SMC as assayed by MTT assay was determined (Table 3) and was
not found to possess significant cytotoxicity.
Table 3: Cytotoxicity of SMC on L6 cells by MTT assay.
Concentration of SMC (µg/ml)
500
250
125
62.5
31.25
15.625
7.813
3.906
1.953
% viability
81.31±0.366
85.50±0.629
87.00±0.301
88.45±0.512
91.03±0.296
91.86±0.442
93.28±0.323
97.10±0.280
98.66±0.240
Values are expressed as mean± SEM. n=8
The glucose uptake activity of SMC was compared with insulin and metformin, and the results are shown in Table 4.
Table 4: In vitro glucose uptake by L6 cells in the presence of SMC, Insulin and Metformin.
% glucose uptake
w.r.t. control
65.58±3.10
86.65±0.18
90.09±1.18
93.96±0.91
114.60±0.88
115.46±2.12
119.76±1.26
149.01±1.56
200.18±0.55
209.21±0.25
SMC (mg/ml)
0.01
0.02
0.03
0.04
0.05
0.06
0.07
0.08
0.09
0.1
% glucose uptake
w.r.t. control
102.56±0.48
107.29±1.21
114.60±2.22
118.04±2.12
127.93±1.26
135.25±0.95
167.07±1.08
184.27±0.58
234.15±1.08
242.75±1.21
DISCUSSION
Management of optimum blood glucose level through the regulation of insulin is of extreme importance in hyperglycemic
disorders like Type I and II diabetes mellitus. Any agent that increase insulin secretion (in case of Type I DM) or aids in the peripheral
utilization of glucose by sensitizing the cells to insulin (in case of Type II DM) is a claimant to be a drug candidate belonging to the
anti-diabetic category. Hypoglycemic herbs increase insulin secretion, enhance glucose uptake by adipose or muscle tissues, and
inhibit glucose absorption from intestine and glucose production from the liver [21]. Crude saponins from M. cymbalaria have been
demonstrated to have antidiabetic and antihyperlipidemic activities. Momordica charantia is reported to contain saponins such as
charantin, momordicine, insulin-like steroidal saponin, and triterpenes saponins, etc., which are responsible for its antidiabetic activity
[22]. Insulin stimulates glucose uptake in skeletal muscle cells and fat cells by promoting the rapid translocation of GLUT4 glucose
transporters to the plasma membrane. It regulates the release of GLUT4 from sequestered intracellular storage pools, and also has
effects on docking and fusion of GLUT4 vesicles with plasma membrane [23]. Regulation of GLUT4 activity by insulin will enhance
the muscle cell glucose uptake. Medicinal plants enhance the glucose uptake by GLUT4 translocation and were proven by in vitro
glucose uptake model [24-27]. The insulin responsiveness of the membrane transport system for glucose (2-deoxy-D-glucose) in
diaphragm was measured during postnatal development of the rat. The results indicate that the extent of insulin stimulation of glucose
www.iajpr.com
2074
Metformin
(mg/ml)
0.01
0.02
0.03
0.04
0.05
0.06
0.07
0.08
0.09
0.1
Page
% glucose uptake
w.r.t. control
0.01
11.40±1.22
0.02
40.21±0.88
0.03
63.00±2.13
0.04
67.73±1.56
0.05
78.91±2.61
0.06
82.78±3.05
0.07
86.65±1.15
0.08
88.80±0.68
0.09
115.89±2.14
0.1
196.74±1.15
Values are expressed as mean± SEM. n=8
Insulin (mg/ml)
Vol 5, Issue 05, 2015.
Mr. Suman Samaddar et al.
ISSN NO: 2231-6876
(2-deoxy-D-glucose) transport in the diaphragm during the first 20 days of life is not directly or simply related to the number of
insulin receptors or the number of intracellular glucose transporters. The extent of the insulin response depends on some other factor
that activates or is part of the machinery for translocation of the transporter [28]. It has also been found that the sulfonylureas
glibenclamide and glimepiride stimulates glucose utilization by rat diaphragm, and this stimulation is correlated with modulations of
the cAMP regulatory cascade [29]. Many phytochemicals have been found to possess glucose uptake activity. Ficus hispida
demonstrated considerable glucose uptake activity by isolated rat hemidiaphragm when compared to glibenclamide [30]. Studies with
Aegle marmelos Correa root and Alpinia calcarata showed similar results [31, 32]. Saponin of M. cymbalaria enhanced glucose uptake
in non-diabetic rat diaphragm when compared to normal control group (p>0.001). The effect is more than insulin group and
comparable to Metformin treated group. In the case of diabetic animals, glucose uptake by SMC has been found to be better than the
insulin and Metformin groups.
In vitro models comprising of skeletal muscle cells and adipocytes are widely used to study glucose uptake activity of drugs.
The L-6 cell line is the best characterized cellular model origin to study glucose uptake and GLUT4 translocation [33, 34]. A 3T3-L1
adipocyte model has been used to study the effect of two sulfonylureas glibenclamide and glimepiride on glucose uptake. It has been
found that they stimulate glucose uptake, and this stimulation is correlated with modulations of the cAMP regulatory pathway [29].
Saponin of Momordica cymbalaria (SMC) was evaluated for its cytotoxicity on L6 cells by MTT assay. The viability was found to be
about 82% at a concentration of 500µg/ml (Table 4). This indicates that SMC does not have significant cytotoxicity on L6 cells.
Glucose uptake by L6 in the presence of SMC, insulin and metformin occurred in a dose-dependent manner. The results show that
uptake in the presence of SMC is higher as compared to metformin and insulin. The uptake peaked to 242.75±1.21% for SMC (0.1
mg) as compared to control. Aldose reductase over-expression has been implicated in the activation of protein kinase C (PKC) which
in turn impairs insulin-mediated glucose uptake [35]. Aldose reductase inhibitors like epalrestat have been found to enhance insulinmediated glucose uptake in vascular smooth muscle cells due to suppression of increased PKC activity [36]. The aldose reductase
inhibitory effect of SMC is still under investigation which will aid in the understanding of its mechanism of action.
CONCLUSION
The findings of this study indicate that saponin of Momordica cymbalaria possesses significant type II antidiabetic activity. It
enhanced glucose uptake by both isolated rat diaphragm (diabetic and non-diabetic) and L6 cells considerably as compared to insulin
and metformin. This may be due to its effect on the number of receptors located in the skeletal muscle cell line or on any mechanism
that tend to suppress insulin-mediated glucose uptake in hyperglycemic state. Elucidation of its exact mechanism of action calls for
further research. The phytoconstituent is a promising drug candidate against insulin-resistant hyperglycemia.
Authors’ Statements
Competing Interests
The authors hereby declare that there is no conflict of interest over the research publication concerned.
ACKNOWLEDGEMENT
The authors thank Karnataka College of Pharmacy, Bangalore, India, for experimental infrastructure and financial support,
and Jawaharlal Nehru Technological University, Anantapur, Andhra Pradesh, India, for technical support.
Page
2075
List of Abbreviations:
SMC – Saponin of Momordica cymbalaria
MTT - 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide
GOD-POD - glucose oxidase-peroxidase
GLUT4 - Glucose Transporter 4
FBS - Fetal Bovine serum
PBS - Phosphate Buffered Saline
DMEM - Dulbecco’s Modified Eagle’s Medium
EDTA – Ethylene Diamine Tetraacetic acid
IAEC - Institutional Animal Ethics Committee
CPCSEA - Committee for the Purpose of Control and Supervision of Experiments on Animals
OPPTS - Office of Prevention, Pesticides and Toxic Substances
PKC - Protein Kinase C
www.iajpr.com
ISSN NO: 2231-6876
REFERENCES
1. Li WL, Zheng HC, Bukuru J, De Kimpe N, Natural medicines used in the traditional Chinese medical system for therapy of
diabetes mellitus. J Ethnopharmacol 2004; 92:1-21.
2. Maritim AC, Sanders RA, Watkins JB, Diabetes, oxidative stress, and antioxidants: a review. J Biochem Mol Toxicol 2003;
17(1):24-38.
3. Shukla R, Sharma SB, Puri D, Prabhu KM, Murthy PS, Medicinal plants for treatment of diabetes mellitus. Indian J Cin Biochem
2000; 15:169-77.
4. Grover JK, Yadav S, Vats V, Medicinal plants of India with antidiabetic potential. J Ethnopharmacol 2002; 81:81-100.
5. Klip A, Ishiki M. Recent developments in the regulation of glucose transporter-4 traffic: new signals, locations, and partners,
Endocrinology 2005; 146:5071–78.
6. Petersen KF, Shulman GI. Etiology of insulin resistance. Am J Med 2006;119:10S–16S
7. Gupta RN, Pareek A, Suthar M, Rathore GS, Bansiwal PK, Jain D, Study of glucose uptake activity of Helicteres isora Linn fruits
in L6 cell lines. Int J Diab Dev Ctries 2009; 29(4):170-3.
8. Kameswararao B, Kesavulu M M, Apparao C. Evaluation of antidiabetic effect of Momordica cymbalaria fruit in alloxan-diabetic
rats, Fitoterpia 2003; 74:7-13.
9. Kameswararao B, Kesavulu M M, Apparao C. Antihyperglycemic activity of Momordica cymbalaria in alloxan diabetic rats, J
Ethnopharmacol 2001; 78(1);67-71.
10. Kameswararao B, Kesavulu M M, Giri R, Apparao C. Antidiabetic and hypolipidemic effect of Momordica cymbalaria Hook fruit
powder in alloxan-diabetic rats, J Ethnopharmacol 1999;67:103-9
11. Raju K & Balaraman R. Antidiabetic Mechanisms of Saponins of Momordica cymbalaria, Phcog Mag 2008; 4(15):197-206.
12. Koneri R, Balaraman R, Saraswati C D. Antiovulatory and abortifacient potential of the ethanolic extract of roots of Momordica
cymbalaria Fenzl in rats, Indian J Pharmacol 2006; 38:111-14.
13. Koneri R, Saraswati C D, Balaraman R, Ajeesha E A. Anti-implantation activity of the ethanolic root extract of Momordica
cymbalaria Fenzl in rats, Indian J Pharmacol 2007; 39:90-96.
14. Koneri R, Balaraman R, Hariprasad, Vinoth Kumar M, Ali A. Cardioprotective effect of Momordica cymbalaria Fenzl in rats with
Isoproterenol-induced myocardial injury, J Clin Diagnos Res 2008; 2:699-705.
15. Koneri RB, Samaddar S, Simi SM, Rao ST. Neuroprotective effect of a triterpenoid saponin isolated from Momordica cymbalaria
Fenzl in diabetic peripheral neuropathy, Indian J Pharmacol 2014; 46(1):76-81.
16. Sharda LD, Somshekhar SK. Isolation and characterization of phytoconstituents from Chlorophytum borivilianum, Pharmacog
Res 2010; 2:343-349.
17. Kade I, Barbosa N. Aqueous extracts of Sphagneticola trilobata attenuates streptozotocin-induced hyperglycemia in rat models by
modulating oxidative stress parameters, Biol Med 2010; 2:1-13.
18. Trinder P. Determination of blood glucose using an oxidase-peroxidase system with a non-carcinogenic chromogen. J Clin Pathol
1969; 22(2):158-161.
19. Freshney Ian R. Cytotoxicity. Culture of animal cells: A manual of basic technique, fifth edition, 2005:359 John Wiley & Sons,
Inc.
20. Hongxiang H, George T, Liang VWG. Hypoglycemic herbs and their action mechanisms, Chin Med 2009; 4:1-11.
21. Jesada P, Sutawadee C, Motonobu G, Weena J, Mitsuru S, Artiwan S. New approach for extraction of charantin from Momordica
charantia with pressurized liquid extraction, Sep Purif Technol 2007; 52(3):416-422.
22. Sophie E, Jeremy M. The molecular basis of insulin-stimulated glucose uptake: signaling, trafficking and potential drug targets, J
Endocrinol 2009; 203:1–18.
23. Sudharshan Reddy D, Srinivasa Rao A, Pradeep Hulikere A. In vitro antioxidant and glucose uptake effect of Trichodesma
indicum in L-6 cell lines, Int J Pharm Bio Sci 2012; 3(4):810–819.
24. Yarasu N, Smana P, Pavankumar R, Nareshchandra RNBS, Vinil Kumar V. In vitro glucose uptake assay of hydro methanolic
leaves extract of Syzygium jambos (L) alston in rat skeletal muscle (L6) cell lines, Indo Am J Pharm Res 2013; 3(9):7336-7341.
25. Anitha Mary M, Sujith K, Santosh P, Christina AJM. Study of glucose uptake activity of Solanum xantohocarpum in L-6 Cell
Lines, Euro J Biol Sci 2013; 5(3):77-81.
26. Premkumar N, Soumya K, Dharsana JN, Seena K, Anjana AK. Comparing the effect of antidiabetic activity of Andrographis
paniculata, Salacia reticulate and Ocimum sanctum by in vitro screening, Asian J Pharm Clin Res 2012; 5(4):146-149.
27. Gupta RN, Pareek A, Rathore GS, Suthar M. Study of glucose uptake activity of Helicteresisora Linn. fruits in L-6 cell lines, Int J
Diabetes Dev Ctries 2009; 29:170-173.
28. Wang C. Insulin-stimulated glucose uptake in rat diaphragm during postnatal development: Lack of correlation with the number of
insulin receptors and of intracellular glucose transporters. Proc. Natl. Acad. Sci. 1985; 82:3621-3625.
29. Muller G, Wied S, Wetekam EM, Crecelius A, Unkelbach A, Punter J. Stimulation of glucose utilization in 3T3 adipocytes and rat
diaphragm in vitro by the sulphonylureas glimepiride and glibenclamide, is correlated with modulations of the cAMP regulatory
cascade, Biochemical Pharmacology 1994; 48(5):985-996.
30. Ghosh R, Sharatchandra K, Rita S, Thokchom IS. Hypoglycemic activity of Ficus hispida (bark) in normal and diabetic albino
rats, Indian J Pharmacol 2004;36(4):222-225.
31. Subban R, Sadashiva CT, Tamizmani T, Balasubramanian T, Rupeshkumar M, Balachandran I. In vitro glucose uptake by isolated
rat hemi-diaphragm study of Aegle marmelos Correa root, Bangladesh J Pharmacol 2009; 4:65-68.
www.iajpr.com
2076
Mr. Suman Samaddar et al.
Page
Vol 5, Issue 05, 2015.
Vol 5, Issue 05, 2015.
Mr. Suman Samaddar et al.
ISSN NO: 2231-6876
32. Rajasekar R, Manokaran K, Rajasekaran N, Duraisamy G, Kanakasabapathi D. Effect of Alpinia calcarata on glucose uptake in
diabetic rats-an in vitro and in vivo model, Journal of Diabetes & Metabolic Disorders 2014; 13(33):1-11.
33. Patel MB, Mishra SH. Cell lines in diabetes, Phcog Res 2008; 2:188-205.
34. Yasunari K, Kohno M, Kano H, Yokokawa K, Minami M, Yoshikawa J. Mechanism of action of troglitazone in the prevention of
high glucoseinduced migration and proliferation of cultured coronary smooth muscle cells, Circ Res. 1997; 81:953–962.
35. Yasunari K, Kohno M, Kano H, Minami M, Yoshikawa J. Aldose reductase inhibitor improves insulin-mediated glucose uptake
and prevents migration of human coronary artery smooth muscle cells induced by high glucose, Hypertension 2000; 35:1092-1098.
36. Ramana KV, Friedrich B, Tammali R, West MB, Bhatnagar A, Srivastava SK. Requirement of aldose reductase for the
hyperglycemic activation of protein kinase C and formation of diacylglycerol in vascular smooth muscle cells, Diabetes 2005;
54:818-829.
Page
2077
54878478451150526
www.iajpr.com