Pharmaceutical Biology, 2011; 49(1): 3237

2011 Informa Healthcare USA, Inc.

ISSN 1388-0209 print/ISSN 1744-5116 online
DOI: 10.3109/13880209.2010.493178

RESEARCH ARTICLE

Antidiabetic effect of Raphanus sativus root juice

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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

fective treatment especially for usage in the developing as

well as under-developed countries. Since synthetic drugs
have undesirable side effects or contraindications, the
World Health Organization (WHO) has recommended
the evaluation of traditional plant treatments for diabetes
(Day, 1998). India is a country with a vast reserve of natural resources and a rich history of traditional medicines
(Grover & Vats, 2001). A number of Indian medicinal
plants have been used for thousands of years in the traditional system of medicine for treating various diseases.
Although medicinal plants have been historically used
for diabetes treatment throughout the world, few of them
have been validated by scientific criteria. The medicinal
properties of plants have now been investigated scientifically throughout the world, due to their potent pharmacological activities, low toxicity, and economic viability

Diabetes mellitus is a global burden as the number of

diabetic patients is increasing year by year. Diabetes is
characterized by a loss of glucose homeostasis resulting
in high blood glucose level (Scheen, 1997). It is the most
common metabolic disorder considered among five leading causes of death in the world (Gispen & Biessels, 2000;
Rahimi et al., 2005). It is a serious illness with multiple
complications and premature mortality. India, China, and
United States will be the countries with the largest number of diabetic patients by the year 2025 (King etal., 1998).
India leads the world with the largest number of diabetic
subjects (Balasubramanyam & Mohan, 2007) earning the
dubious distinction of being termed the diabetes capital
of the world (Mohan et al., 2007). Therefore, it has become necessary to look for novel oral therapeutically ef-

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

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Antidiabetic effect of Raphanus sativus 33

(Singh etal., 2008; Gupta etal., 2005). The ethnobotanical
information reports on approximately 800 plants that may
possess antidiabetic potential (Alarcon-Aguilara et al.,
1998). Several such herbs have shown antidiabetic activity when assessed using experimental techniques (Saifi
etal., 1971; Mukherjee etal., 1972; Coimbra etal., 1992;
Kar etal., 1999; Jafri etal., 2000). In recent years, several
plants commonly used to treat diabetes in the traditional
system of medicine have been explored scientifically by
our research group for investigating their chemical constituents, elemental analysis, their role in diabetes management, and pharmacological activities (Kesari et al.,
2004; Rai etal., 2007a, 2007b, 2008).
Radish [Raphanus sativus L. (Brassicaceae)] is most
valued by the inhabitants of many Western and Eastern countries as a food and medicine (Mayer, 1981).
In Greeko-Arab or Unani medicine as well as in Indian
folklore, radish is administered as a household remedy
for the prevention and treatment of gall stone, jaundice,
flatulence, indigestion, and in various gastric ailments.
Eating a few slices of raw radish with salt and pepper,
three times daily, decreases complaints of piles, constipation, indigestion, colic, dyspepsia, enlargement of liver,
spleen, jaundice, and prolapse of the rectum (Prahoveanu & Esanu, 1990). Radish extract has been found to
stimulate gastrointestinal mobility in rodents (Jung etal.,
2000). The juice of radish has a tonic and laxative action
on the intestine and indirectly stimulates the flow of bile
(Aman, 1969). In addition, radish was found to contain
indole-3-carbinol which exhibited strong hepatoprotective properties against various carcinogenic agents (Aggarwal & Ichikawa, 2005). Acylated anthocyanins have
been isolated from the root peels, petioles, and flowers
of Raphanus sativus (Otsuki etal., 2002; Tatsuzawa etal.,
2008). Polyadenylic and polyadenylated ribonucleic
acids have been isolated from R. sativus (Aspart et al.,
1979). The protective effects of aqueous extract of R.
sativus against zearalenone-induced reproductive toxicity and oxidative stress in mice have also been reported
(Salah-Abbs etal., 2009). The suppression of lipid peroxidation and oxidative DNA damage in rats have been
experimentally proven by the intake of R. sativus indicating thereby that the vegetable possesses an antioxidative
effect in vivo which could be related to the prevention of
carcinogenesis (Ippoushi etal., 2007). Thus, the present
investigation was undertaken to evaluate the glycemic
profile of the Raphanus sativus root juice on blood glucose level (BGL) of normal and streptozotocin (STZ)induced sub- and mild-diabetic rats during fasting blood
glucose (FBG) and glucose tolerance test (GTT) studies,
so that a novel oral antidiabetic agent could be identified
with high nutritive value.

Methods
Chemicals
Streptozotocin was purchased from Sigma-Aldrich,
Seelze, Germany. Blood glucose level (BGL) for fasting
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blood glucose (FBG) and glucose tolerance test (GTT)

studies was assayed using kits, from Bayer Diagnostics,
New Delhi, India and a one touch Accu-Chek sensor from
Roche Diagnostics, Mannheim, Germany. The solvents
were from Merck, Darmstadt, Germany.

Preparation of crude drug

Fresh roots of Raphanus sativus, about 10kg, were
purchased in June, 2009 from the local market of Allahabad, U.P. (India), and authenticated by Satya
Narayan, Taxonomist, Department of Botany, University of Allahabad, India. A voucher specimen has
been submitted to the university herbarium. The material was washed and squeezed in an electric blender
to obtain fresh juice. 2L of this juice was filtered and
concentrated in rotary evaporator at 35 5C under
reduced pressure to obtain a semisolid material (353g)
which was then lyophilized to get a powder (40g,
11.3%, w/w).

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

34 Surekha Shukla etal.

based on similar studies of FBG and GTT (Rai etal., 2008;
Mehta etal., 2009).

Evaluation of glycemic management in normal

healthy rats

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Six groups of six rats each fasted overnight were used

in the experiment; group I serving as untreated control
received vehicle (distilled water) only, whereas the animals of groups II, III, IV, and V received lyophilized juice
suspended in distilled water at doses 100, 200, 300, and
400mgkg1, respectively. FBG was checked by collecting
blood samples from the tail vein at 2, 4, and 6h after administering the juice.

Assessment of hypoglycemic activity by GTT in normal

healthy rats
The juice was given orally to different groups of overnight
fasted normal healthy animals in the same fashion as
above and the FBG was checked at 2h and treated as 0h
value for GTT. The animals were then orally treated with
4gkg1 of glucose and their glucose tolerance was studied
at 1h intervals for another 3h. Thus, the total period of
blood collection was up to 5h.

Study of antidiabetic activity by GTT in sub- and

mild-diabetic rats
The antidiabetic effect of Raphanus sativus root juice
in sub- and mild-diabetic rats was also assessed by improvement in glucose tolerance. The rats were divided
into six groups. Group I control, received vehicle (distilled
water) only, whereas variable doses of 100, 200, 300, and
400mgkg1 of juice extract were given orally to groups
II, III, IV, and V, respectively. Blood glucose levels were
first checked after 90min of treatment, considered as 0h
value, and then 2gkg1 glucose were given orally to all
the groups. Blood glucose levels were further checked up
to 3h at regular intervals of 1h each, considered as 1, 2,
and 3h values. The results were compared with group VI
rats, which were treated with 2.5mgkg1 of glibenclamide
(synthetic hypoglycemic agent).

LD50 experiment

The toxic effect of the Raphanus sativus root juice was

also studied by a LD50 experiment. Two groups of rats of
both sexes (six animals per group, three females and three
males), weighing about 180200g, were orally treated

with a single dose of 2 and 3g of the Raphanus sativus

root juice. Then, rats were observed for gross behavioral,
neurologic, autonomic, and toxic effects continuously.
Food consumption, feces and urine were also examined
at 2h and then at 6h intervals for 24h.

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.

Effect on FBG in normal rats during GTT

Table 2 deals with the study of Raphanus sativus root
juice on BGL levels and glucose tolerance of normal healthy rats. Different doses of 100, 200, 300, and
400mgkg1 of the juice were given orally to overnight
fasted healthy rats. The FBG was checked after 2h considered as 0h value and then 4gkg1 of glucose was given.
The fall was observed up to 3h after glucose administration at 1h intervals and the results reveal that the percentage fall in BGLs was regular up to the dose of 300mgkg1
and reached its maximum at 15.8%. Moreover, the fall of
10.1, 11, and 14.6% was observed with the dose of 100,
200, and 400mgkg1 at the same time.

Effect on diabetic rats during GTT

Figures 1 and 2 demonstrate the antidiabetic effect of the
Raphanus sativus root juice on mildly and sub-diabetic
animals, respectively. Different doses of the juice as
mentioned above and the standard drug, glibenclamide
(2.5mgkg1) were given orally to the groups as defined
in the experimental design. A fall of 17, 18.7, 23.8, and
22.4% in BGLs of sub-diabetic animals was observed after 3h of glucose administration with doses of 100, 200,

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*

*p<0.05 as compared with control.



Pharmaceutical Biology

Antidiabetic effect of Raphanus sativus 35

Table 2. Effect of variable doses of Raphanus sativus root juice on BGL during GTT of normoglycemic rats (mean SD).
Blood glucose levels (mg dL1)
Post-treatment (h)
Pre-treatment
Treatment
FBG
0
1
2
Experimental Groups
(mg kg1bw)
Control
Distilled Water
90.33.5
90.14.3
108.53.9
107.54.1
Treated 1
100
91.43.6
90.03.3
105.43.5
98.34.3
Treated 2
200
90.53.9
88.93.7
104.34.0
97.53.4*
Treated 3
300
89.53.4
87.13.9
100.53.4
93.63.3*
Treated 4
400
90.74.1
88.34.2
101.23.6
93.73.4*
DW, distilled water.
*p<0.05 as compared with control.

BGL (mg/dL)

200

**

150

**

**

100

animals with the doses of 100, 200, 300 and 400mgkg1,

respectively. However, the dose of 2.5mgkg1 of glibenclamide reduced BGL by 27.2% at 3h during GTT in
mild-diabetic rats, which is less effective than the dose
of 300mgkg1of the juice.

LD50

50
0
FBG

0h

1h
Time (h)

2h

3h

Control

Treated 1

Treated 2

Treated 3

Treated 4

Glibenclamide

**p<0.01 as compated with control: Distilled water, Treated 1: 100 mg kg1,

Treated 2: 200 mg kg1, Treated 3: 300 mg kg1, Treated 4: 400 mg kg1,
Glibenclamide: 2.5 mg kg1

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

**p<0.01 as compated with control: Distilled water, Treated 1: 100 mg kg1,

Treated 2: 200 mg kg1, Treated 3: 300 mg kg1, Treated 4: 400 mg kg1,
Glibenclamide: 2.5 mg kg1

Figure 2. Effect of variable doses of Raphanus sativus juice on BGL

during GTT in mid-diabetic rats.

300, and 400mgkg1, respectively. However, the dose of

2.5mgkg1 of glibenclamide reduced BGL by 24% at 3h
during GTT in sub-diabetic rats. Thus, the results clearly
reveal that the effects of both the dose of 300mgkg1 of
juice and the dose of 2.5mgkg1 of glibenclamide are
almost the same during GTT in sub-diabetic rats. Moreover, the fall observed after 3h of glucose administration
was 10.7, 17.3, 28.3, and 25.7% in BGLs of mild-diabetic
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The experiment was carried out on normal healthy rats.

The behavior of the treated rats appeared normal. No
toxic effect was reported at doses up to 10- and 15-times
the effective dose of the juice and there was no death in
any of these groups.

Discussion

Figure 1. Effect of variable doses of Raphanus sativus juice on BGL

during GTT in sub-diabetic rats.

BGL (mg/dL)

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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

36 Surekha Shukla etal.

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reach to the target tissues through circulation to exhibit

their hypoglycemic and antidiabetic effect which remains significant even after 3h of glucose administration. The dose of 300mgkg1 of the Raphanus sativus
juice was found to be more effective than the dose of
2.5mgkg1 glibenclamide in the case of mildly diabetic
rats, whereas in the case of sub-diabetic rats, the effect
of the same dose of the juice as well as that of glibenclamide is practically the same. Moreover, the effectiveness of the juice in both the cases of mildly as well as
sub-diabetic rats is comparable with the synthetic drug
glibenclamide.

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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