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Jurnal Lobak Diabetse
Jurnal Lobak Diabetse
RESEARCH ARTICLE
Surekha Shukla, Sanjukta Chatterji, Shikha Mehta, Prashant Kumar Rai, Rakesh Kumar Singh,
Deepak Kumar Yadav, and Geeta Watal
Alternative Therapeutics Unit, Drug Development Division, Medicinal Research Lab, Department of Chemistry,
University of Allahabad, Allahabad, India
Abstract
Context: Many plants have been explored scientifically and systematically and claimed to be useful for the treatment
of diabetes mellitus by various research groups worldwide. The present study is a further effort in the direction of
developing a novel oral antidiabetic agent of high potential with minimal or no side effects.
Objective: This study screened the glycemic attributes of Raphanus sativus L. (Brassicaceae) root juice in normal as well
as sub- and mild-diabetic models.
Materials and methods: The variable doses of 100, 200, 300, and 400mgkg1 body weight (bw) of the extract were
administered orally to normal and streptozotocin (STZ)-induced sub- and mild-diabetic rats in order to define its
glycemic potential. Glibenclamide was used as a reference drug.
Results: The dose of 300mgkg1bw was identified as the most effective dose which lowers the blood glucose level
(BGL) by 33.4% (p<0.001) at 6h during fasting blood glucose (FBG) studies in normal rats. However, the glucose
tolerance test (GTT) revealed the maximum reduction of 15.9% (p<0.001) in BGL at 3h in normal rats with the same
dose, whereas the reduction observed was by 23.8 and 28.3% (p<0.001) in sub- and mild-diabetic rats, respectively,
at the same interval of time.
Discussion and conclusion: This evidence clearly indicates that Raphanus sativus root juice possesses good hypoglycemic
potential coupled with antidiabetic efficacy.
Keywords: Antidiabetic, glucose tolerance test, glibenclamide, hypoglycemic, Raphanus sativus, streptozotocin
Introduction
Address for Correspondence: Geeta Watal, Alternative Therapeutics Unit, Drug Development Division, Medicinal Research Lab, Department
of Chemistry, University of Allahabad, Allahabad, India. Tel.: +91-532-2462125; E-mail address: geetawatal@gmail.com
(Received 22 December 2009; revised 08 April 2010; accepted 11 May 2010)
32
Methods
Chemicals
Streptozotocin was purchased from Sigma-Aldrich,
Seelze, Germany. Blood glucose level (BGL) for fasting
2011 Informa Healthcare USA, Inc.
Experimental animals
More than 100 male albino Wistar rats of the same age
group and body weight 150200g were selected for the
experiments. Animals obtained from the National Institute of Communicable Disease (NICD), New Delhi, India
were housed in polypropylene cages at an ambient temperature of 25-30C and 45-55% relative humidity with
a 12h each dark and light cycle. Animals were fed pellet
diet (Pashu Aahar Kendra, Varanasi, India) and water ad
libitum. The study was approved by the Institutional Ethical Committee.
Induction of diabetes
Diabetes was induced to overnight fasted rats by a single
intraperitonial injection of freshly prepared streptozotocin (STZ) 50mgkg1bw in 0.1M citrate buffer (pH=4.5)
(El-Fiky et al., 1996). After 3 days of STZ administration, rats with marked hyperglycemia were selected
for the study (Rai et al., 2008). The rats with hyperglycemia were divided into two groups of 36 rats each:
sub-diabetic animals with normal FBG (80-120)mgdL1
and abnormal postprandial (PPG>210mgdL1), and
mild-diabetic animals with FBG 150200mg dL1and
PPG>250mg dL1.
Estimation
Blood glucose level (BGL) was estimated by the glucose
oxidase method (Barham & Trinder, 1972) using a standard kit from Bayer Diagnostics India.
Experimental design
Initial screening of the Raphanus sativus root juice for the
hypoglycemic activity was done with a range of variable
doses in normal healthy rats by conducting fasting blood
glucose (FBG) and glucose tolerance test (GTT) studies. The antidiabetic effect was assessed in sub- as well
as mild-diabetic models with the same range of doses
LD50 experiment
Statistical analysis
Data were statistically evaluated using two-way ANOVA,
followed by a post hoc percentage considered significant
when p<0.05.
Results
Effect on FBG in normoglycemic rats
Table 1 summarizes the hypoglycemic effect of a single
oral treatment of variable doses of 100, 200, 300, and
400mgkg1 of root juice in normal healthy rats. Treated
rats showed a regular fall of 8.9, 14.8, and 33.4% from
the doses of 100, 200, and 300mgkg1, respectively, after
6h. However, a fall of only 29.9% was observed with an
increased dose of 400mgkg1 after the same interval of
time.
Table 1. Effect of variable doses of Raphanus sativus juice on BGL during FBG test of normoglycemic rats (mean SD).
Blood glucose levels (mg dL1)
Post-treatment (h)
Pre-treatment FBG
2h
4h
Experimental Groups
Treatment (mg kg1bw)
Control
Distilled water
98.83.4
98.33.9
97.73.3
Extract
100
95.63.2
94.14.1
91.33.5
Extract
200
94.43.9
84.73.7
82.64.0
Extract
300
98.53.1
81.23.9
75.84.1
Extract
400
96.53.8
83.04.2
78.04.3
6h
96.73.7
88.13.4*
80.43.6*
65.53.8*
67.53.9*
Pharmaceutical Biology
BGL (mg/dL)
200
**
150
**
**
100
LD50
50
0
FBG
0h
1h
Time (h)
2h
3h
Control
Treated 1
Treated 2
Treated 3
Treated 4
Glibenclamide
400
350
300
250
200
150
100
50
0
**
FBG
0h
1h
Time (h)
**
**
2h
3h
Control
Treated 1
Treated 2
Treated 3
Treated 4
Glibenclamide
Discussion
BGL (mg/dL)
250
3
107.73.7
96.23.6*
95.33.7*
90.13.5*
91.43.8*
Raphanus sativus root juice contains essential oils (Popovic etal., 1993). It has been used in Indian traditional
system of medicine but no data has been reported so
far for its glycemic profile in vivo. Hence, this study
deals with complete screening of its glycemic attributes
based on FBG and GTT studies in normal as well as
STZ-induced diabetic animal models, respectively.
The observed difference between initial and final BGLs
of different groups of animals during these studies revealed a significant elevation in blood glucose in the
control group as compared with treated groups. Table
1 indicates that the maximum hypoglycemic effect was
produced at 6h with a subsequent rise in blood glucose
level at 8h during FBG studies in normal rats. Table 2
reveals the maximum hypoglycemia at 5h in glucose
loaded rats on oral administration of Raphanus sativus root juice. It is generally accepted that the sulfonyl
ureas, including glibenclamide, produce hypoglycemia
in normal as well as diabetic animals by stimulating the
pancreatic -cells to release more insulin (Goth, 1985;
Larner, 1985). Hence, the significant reduction as shown
in BGLs of diabetic rats treated with the Raphanus sativus juice as well as glibenclamide (Figures 1 and 2)
may be due to stimulation of the residual pancreatic
mechanism, probably by increasing peripheral utilization of glucose as postulated by Earth etal. (1996). This
validates the efficacy of the juice to control elevated
blood sugar levels. However, the blood sugar levels were
maintained in normal and diabetic rats throughout the
period of study. These data suggest the active ingredients of the juice or their metabolites need about 2h to
Conclusion
Thus, it has been scientifically proven that the Raphanus
sativus root juice has significant hypoglycemic as well
as antidiabetic potential. The antidiabetic effect of the
juice was even greater than the drug glibenclamide. Enzymatic studies are in progress in order to elucidate the
detailed mechanism of action at cellular and molecular
levels. Isolation and characterization of compounds
of juice responsible for lowering of BGL is also under
investigations.
Declaration of interest
The first author (S.S.) is grateful to the UGC (University
Grants Commission), Government of India, for financial
assistance and is thankful to the Government of India for
providing a fellowship. The authors report no conflicts of
interest. The authors alone are responsible for the content and writing of the paper.
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