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R. Arun kumar* et al. /International Journal Of Pharmacy&Technology
ISSN: 0975-766X
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COMPARATIVE EVALUATION OF EXTRACTS OF C. IGNEUS (OR C. PICTUS) FOR
HYPOGLYCEMIC AND HYPOLIPIDEMIC ACTIVITY IN ALLOXAN DIABETIC RATS.
Panagal Mani1, R. Arun kumar*2, T.M.M.John Bastin3, S.jenifer3, Muthuvel Arumugam4,
1
Department of Zoology and Biotechnology, A.V.V.M Sri Pushpam College, Poondi,
Thanjavur-613 503, Tamil Nadu, India.
2
PG and Research Lab, Department of Biotechnology, Marudhu Pandiyar College, Thanjavur-
613 403, Tamil Nadu, India.
3
PG and Research Lab, Department of Biotechnology, Tamilvel Umamaheswaranar Karanthai
ArtsCollege, Thanjavur- 613002, Tamil Nadu, India.
4
Centre of Advanced Study in Marine Biology, Annamalai University, Parangipettai – 608 502,
Tamilnadu, India.
Received On: 10-02-2010 Accepted On: 25-02-2010
Abstract
Aims: To compare the possible therapeutic effects of C. igneus methanolic and aqueous extracts
against hyperglycemic and hyperlipidemic activity in alloxan induced diabetic rats.
Materials and Methods: Blood glucose level, serum lipid profile and pancreas histology as well as
phytochemical investigations were estimated after administration of the both extracts at dose level of
50, 100 and 200 mg/kg were comparable to that of glibenclamide (0.5mg/kg).
Result: All studies in aqueous and methanolic extracts of Costus igneus have shown better effect on
end of 30th day. The significantly reduce type 2 diabetes in rats and regulation of insulin secretion,
islet protection as well as produce higher antidiabetic compounds by methanolic extract at dose level
of 200 mg/kg.
Conclusion: Our results suggest that the Costus igneus methanolic extract anti-diabetic activity is
slightly higher to the aqueous extract and good agreement with the folk medicinal use of this plant.
Key words: Costus igneus, Flavonoids, Glibenclamide, Hypolipidaemic, Hypoglycemic.
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1. Introduction
Diabetes mellitus is a chronic metabolic disease which now afflicts 3% of the world population (1).
Current anti-diabetic drugs usually have adverse side effects and therefore, there is a need to find
safer and more effective antidiabetic drugs (2). Now-a-days herbal treatments are becoming
increasing by popular as the herbal preparations have no or least side effects (3). In developing
countries, the World Health Organization (WHO) estimates that about 80% of the population relies
on plant based preparations used in their traditional medicinal system and as the basic needs for
human primary health care (4).
Costus igneus belongs to the Costaceae family has been used to treat diabetes. Similar result also
posses that hypoglycemic potential of C. igneus leaves (5) and diosgenin in rhizomes (6). The
purpose of the present study is to compare the hypolipidemic and hypoglycemic effects of
methanolic and aqueous extracts of Costus igneus dried leaves in alloxan induced rats. This study is
based on their blood glucose level along with evaluated the serum total cholesterol, serum
triglycerides, serum LDL, serum VLDL, and serum HDL levels. Phytochemical nature of the dried
leaves extracts has also been carried out based on qualitative chemical tests.
2. Materials and Methods
2.1 Preparation of plant extract
Fresh plant material of C. igneus was purchased from local horticultural farm and various nurseries
and identified to confirm by the Taxonomist. Leaves were cut into pieces, shade dried and powered.
This leaves powder was continuously extracted successively with methanol and distilled water using
soxhlet upto 48 h. Each extracts were filtered and concentrated in rotatory evaporator at 35-40ºC
under reduced pressure to obtain a semisolid material, which was then lyophilized to get powders
(yield 28.4% and 26.9% w/w).
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2.2 Experimental Animals
Male Wistar albino rats, weighing about 150-200 g obtained from the Periyar College of
Pharmaceutical Science, Trichy, Tamil Nadu, India, were used for the study. Animals were housed
under standard conditions of temperature (24±2°C) and relative humidity (30-70%) with a 12:12
light: dark cycle, fed with standard pellet diet (Chakan Oil Mills, Sangli) and water ad libitum.
Ethical clearance was given by Institutional Animal Ethics Committee and conducted experimental
rules of Indian National Science Academy (CPCSEA/265).
2.3 Induction of diabetes in rats
The rats were administered with single intraperitoneal injection of 150mg/kg body weight of alloxan
monohydrate (Sigma, USA), dissolved in normal saline. Two days after alloxan injection, animals
with blood glucose greater than 250 mg/dl were separated and used for the study.
2.4 Experimental design
All the animals were randomly divided into the 9 groups with 7 animals in each group. First 3
groups were served as normal, diabetic, and glibenclamide (0.5 mg/kg) control, respectively. Where
as, groups 4, 5, 6 and 7, 8, 9 diabetic rats were received methanolic and aqueous extracts of C.
igneus (50, 100 and 200 mg/kg) and served as positive control. All the animals then received glucose
solution (3 g/kg) orally and these treatments continued for 30 days respectively. However, the
extracts were administrative intra peritoneally.
2.5 Biochemical assays
From all groups of rats (normal, diabetic control, extracts and standard treated) blood samples were
collected from rats at 10 days interval by cardiac puncture under light ether anesthesia, blood
glucose was analyzing through the glucometer (Johnson & Johnson Co., USA), (7). While, serum
cholesterol (COGENT kit, Span Diagnostic Ltd, India), triglycerides (ENZOPAK kit, Reckon
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Diagnostic Ltd, India) (8), HDL (COGENT kit, Span Diagnostic Ltd, India) (9) LDL and VLDL
(Semi auto analyzer, SECOMAM, France) were measured by using the standard calculation method.
2.6 Histopathological studies
Animals were sacrificed on 10, 20 and 30th day during prolonged treatment. Pancreases were
removed, kept in 10% formaline and immediately processed by the paraffin technique. In
histological examination, sections of .5 µ thickness were mounted on slides and stained with
hematoxylin and eosin. Stained sections were morphologically evaluated.
2.7 Phytochemical investigation
Methanol and aqueous extracts of C. igneus phytochemical analysis assessed the presence of
flavonoids, tannins, saponins, alkaloids, glycosides, lignins and phytosterol by using HPLC protocol
in Soil Testing Laboratory, Trichy.
2.8 Statistical analysis
All data were expressed as means ± S.E.M. Student ‘t’ test was used to compare the mean
values of test groups and control. Differences in mean values were considered significant at p < 0.01.
3. Results
3.1 Blood glucose level
In Table 1, blood glucose levels in methanolic and aqueous extracts of C. igneus treated groups
showed no significant differences at the end of 10th day, but those groups were lower than that in the
diabetic control group after 20th day of treatment. On 30th day of experiment, blood glucose levels in
the methanolic and aqueous extracts treated groups significantly decreased 91.2 and 95.8 mg/dl, by
their high dose 200 mg/kg respectively. The doses 50, 100 mg/kg treated groups does not shows any
significant variation on the blood glucose level throughout the experimental period as compared to
standard drug glibenclamide (83.2 mg/dl).
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Table 1: Effect of methanolic and aqueous extracts of C. igneus leaves on the blood glucose level in rats.
Experimental groups (mg/dl)
Treatment
days Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8 Group 9
control Diabetic Glibenclamid Methanol Methanol Methanol Aqueous Aqueous Aqueous
control e extract (50 extract (100 extract (200 extract (50 extract (100 extract (200
control mg/kg) mg/kg) mg/kg) mg/kg) mg/kg) mg/kg)
Day 1 91.3 ± 4.5 286.6± 2.9 148.6 ± 6.2 228.4±4.2 142.6 ± 4.8 121.6 ± 3.2 234.2 ± 6.2 241.3 ± 5.1 132.4 ±9.1
Day 10 98.0 ± 3.8 282.5± 3.7 98.6 ± 5.1 161.2 ± 8.9 137.4 ± 1.8 109.3 ± 4.1 172.3 ± 2.6 143.0 ± 3.2 112.3 ±3.5
Day 20 86.6 ± 4.1 290.7± 4.1 92.0 ± 4.8 153.9 ± 7.1 126.6 ± 1.9 98.0 ± 7.8 161.7 ± 3.9 131.8 ± 2.7 101.6±1.8
Day 30 95.0 ± 2.5 295.0± 3.3 83.2 ± 3.7 140.3 ± 1.6 103.8 ± 2.5 91.2 ± 6.7 152.0 ±4.6 118.7 ± 5.7 95.8 ± 1.4
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3.2 Effect of LDL on diabetic rats
In Figure 1, LDL levels of normal rats selected for the study were in the range of 72.66 mg/dl
and it was further increased to 198.30 mg/dl by induced diabetes on 30th day. While, the diabetic
rats treated with 200 mg/kg of methanolic and aqueous extracts of C. igneus decreased LDL
levels steadily to 81.06 mg/dl and 95.77 mg/dl respectively, which was comparable to that of
glibenclamide (61.30 mg/dl).
Fig 1: LDL level
3.3 Effect of VLDL on diabetic rats
In Figure 2, the normal VLDL level 15.04 mg/dl was increased upto 37.80 mg/dl by induced
diabetes. Whereas, higher dose treatment of methanolic and aqueous extracts of C. igneus (200
mg/kg) were highly decreased VLDL level upto 18.04, 22.71 mg/dl at end of 30th day was
compared to that of standard drug glibenclamide (14.92 mg/dl) treated diabetic rats.
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Fig 2: VLDL level
3.4 Effect of Triglycerides on diabetic rats
In Figure 3, the normal level of triglycerides was found to be 76.07 mg/dl, which was extremely
increased upto 134.00 mg/dl by alloxan treatment. After treatment, the triglycerides levels were
reduced upto 92.01, 96.58mg/dl by significant dose (200 mg/kg) reaction of methanolic and
aqueous extracts of C. igneus at end of 30th day. The higher dose of glibenclamide reduced
triglycerides level to 73.05 mg/dl.
Fig 3: Triglyceride level
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3.5 Effect of Total cholesterol on diabetic rats
In Figure 4, the normal level of total cholesterol 122.15 mg/dl was increased upto 273.19 mg/dl
by induced diabetes. Whereas, higher dose treatment of methanolic and aqueous extracts of C.
igneus (200 mg/kg) were highly decreased total cholesterol level upto 121.00, 136.14 mg/dl at
day 30 in diabetic rats, it was compared to that of standard drug glibenclamide (96.37 mg/dl).
Fig 4: Total Cholesterol Level
3.6 Effects HDL on diabetic rats
In Figure 5, diabetic animal models were exposed HDL level during hyperlipidemic. The normal
HDL level is 56.06 mg/dl; it was highly decreased upto 21.37 mg/dl by induction of diabetes.
The oral administrations of 200 mg/kg of C. igneus methanolic and aqueous extracts were
significantly improved HDL level upto 57.29, 51.17 mg/dl respectively. Glibenclamide treated
rats showed 63.20 mg/dl of HDL level at the end of 30th day.
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Fig 5: HDL level
3.7 Pancreas histology
Multiple section of pancreas were taken and studied for any histological changes. The pancreas
present in the group of animals treated with the higher doses of methanolic and aqueous extracts
of C. igneus (200 mg/kg) clearly showed that the partial restoration of normal cellular population
and enlarged size of β beta cells with hyperplasia on 30th day. The islets were normal in size,
shape and number comparatively similar to that of standard treated (Figure 6: A, B, C, and D).
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Figure- 6. Photomicrographs rat pancreas stained by haematoxylin and eosin of untreated (A)
and alloxan-induced diabetic (B) rats and effects of aqueous extract (C) methanolic extract (D)
of C. igneus leaves and glibenclamide(E).
3.8 Phytochemical analysis
In phytochemical analysis, methanolic extract of C. igneus produced higher concentration of
flavonoids, alkaloids, tannins, lignins, glycosides and presence of saponins, phytosterol than the
aqueous extract.
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4. Discussion
In the present study, the anti-diabetic activity of methanol and aqueous extracts of C. igneus
leaves was determined. Diabetes mellitus is a collection of disorders, which results from either
lack of insulin or factors, which interfere with the action of this hormone (10). In animals, it can
be induced by partial pancreatectomy or by the administration diabetogenic drugs such as
alloxan, streptozotocin, ditizona and anti-insulin serum (11). Now-a-day alloxan is widely used
to induce diabetes in experimental animals and it causes a massive reduction in insulin release by
the destruction of β-cells of the islets of langerhans and thereby induces hyperglycemia (12).
Present study indicates that the oral administration of methanolic and aqueous extract of C.
igneus higher doses (200 mg/kg) and glibenclamide (0.5 mg/kg) treated groups shows significant
reduction in blood glucose against diabetes such as 91.2, 95.8, 83.2 mg/dl on the 30th day in
alloxan induced rats. Hence, the methanolic extract was found to be almost significant as
standard drug in lowering blood glucose level, whereas the aqueous extract treated groups
showed blood glucose level that is comparatively less to methanol extract (200 mg/kg) and
standard drug.
In hypolipidemic action, an increased risk of coronary heart disease is associated with a high
serum concentration of total cholesterol, low-density lipoprotein, very low-density lipoprotein
and triglyceride. On the other hand, low serum concentration of high-density lipoprotein is also
responsible for coronary heart diseases. However, methanolic and aqueous extract at dose of 200
mg/kg has reversed the diabetes-induced hyperlipidemia compared to their standard drug.
Significant reduction of total cholesterol, triglycerides, LDL, VLDL, and increased HDL in
methanolic extract (200mg/kg) treated was comparative to standard drug treated groups and
reached high value compared to aqueous extract. In histopathological study, reversed
abnormalities are seen in the pancreas of diabetic animals at after treatment with higher doses
(200 mg/kg) of methanolic and aqueous extracts as well as considerable improvement in β-cell
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size with hyperplasia were comparison to that of glibenclamide indicates the anti-diabetic
potential of this plant. Moreover, fall produced normal doses (50 and 100 mg/kg) of both extracts
not possess significant anti-diabetic activities when compared to glibenclamide.
5. Conclusion
The results of the present investigation clearly indicate that the comparing the methanolic and
aqueous extracts of C. igneus shows a better activity to possess possible usefulness in the
treatment of diabetes. Among these two extracts, methanolic extract possess significant
hypoglycemic, hypolipidaemic activity and evaluate important antidiabetic compound in
phytochemical investigation. Hence, a great deal of our research gives a solid scientific approach
to the traditional uses of these medicinal species effects against diabetes mellitus.
Reference
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Corresponding author:
R. Arun Kumar,
Department of Biotechnology,
Marudhu Pandiyar College,
Thanjavur-613 403,
Tamil Nadu, India.
Email: arunkumarkarthi@yahoo.com.
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