Pharmaceutical Biology
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Antidiabetic and Antihyperlipidemic Effects of
Myristica fragrans. in Animal Models
D.K. Arulmozhi, R. Kurian, A. Veeranjaneyulu & S.L. Bodhankar
To cite this article: D.K. Arulmozhi, R. Kurian, A. Veeranjaneyulu & S.L. Bodhankar (2007)
Antidiabetic and Antihyperlipidemic Effects of Myristica�fragrans. in Animal Models, Pharmaceutical
Biology, 45:1, 64-68, DOI: 10.1080/13880200601028339
To link to this article: https://doi.org/10.1080/13880200601028339
Published online: 07 Oct 2008.
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Pharmaceutical Biology
2007, Vol. 45, No. 1, pp. 64–68
Antidiabetic and Antihyperlipidemic Effects of Myristica fragrans
in Animal Models
D.K. Arulmozhi, R. Kurian, A. Veeranjaneyulu, and S.L. Bodhankar
Department of Pharmacology, Bharati Vidyapeeth, Poona College of Pharmacy, Pune, Maharashtra, India
Abstract
The effect of the hydroalcoholic extract of fruits of
Myristica fragrans Houtt. (Myristicaceae) was investigated on chlorpromazine-induced glucose and triglyceride elevations in male Swiss albino mice. After 7 days
of oral administration, the extract, at doses of 150 and
450 mg=kg, ameliorated the metabolic abnormalities
caused by chlorpromazine as evidenced by significant
reduction of glucose and triglyceride (TG) levels (maximal effect of 41% and 53% reduction of glucose and
TG, respectively, at 450 mg dose, p < 0.01). The standard antidiabetic rosiglitazone at 10 mg significantly
(p < 0.01) reduced the TG (63%) and glucose (40%)
levels in this model, while the standard antidiabetic glimepiride has exhibited 55% and 16% reduction in TG
and glucose, respectively. In rats fed a high-cholesterol
diet, Myristica fragrans extract significantly reduced the
elevated TG (47% reduction at 450 mg, p < 0.01) and
cholesterol (66.7% reduction at 450 mg, p < 0.01), and
also exhibited a reduction in hepatic TG secretion after
tyloxapol administration. These data suggest that
Myristica fragrans extract ameliorates hyperglycemia
and abnormal lipid metabolism in animal models.
Keywords: Diabetes, high-cholesterol diet, lipid abnormalities, mice.
Introduction
Type 2 diabetes mellitus (T2DM) is a major and growing
health problem throughout the world. T2DM results from
both peripheral insulin resistance and impaired insulin
secretion. Insulin resistance arises as a consequence of
obesity, a sedentary lifestyle, and aging, with resulting
hyperglycemia and diabetes, blood pressure elevation,
and dyslipidemia, collectively called ‘‘metabolic syndrome
X’’ (Arulmozhi & Portha, 2006). A worldwide epidemic of
obesity, insulin resistance, and diabetes is one of the major
risk factors responsible for the ever-increasing incidence of
heart failure. Evidence from human and animal studies
suggests that lipid accumulation in the heart, skeletal muscle, pancreas, liver, and kidney plays an important role in
the pathogenesis of heart failure, obesity, and diabetes.
Cardiac lipid accumulation induces lipototoxicity, predisposing the myocytes to death and contractile dysfunction
(Zhou et al., 2000; Chiu et al., 2001).
Currently available therapies for diabetes include
insulin and various oral antidiabetic agents, such as sulfonylureas, metformin, a-glucosidase inhibitors, and the
thiazolidinedione class of glitazones. Each of these oral
agents can cause a number of serious side effects (Zang &
Moller, 2000). Hence, there is an imperative need for better
therapeutic agents with fewer side effects for the pharmacotherapy of type 2 diabetes. Plants have provided usable
sources of drugs, and many drugs are directly or indirectly
derived from plants. Phytomedicines have three distinct
advantages: (1) Botanical extracts can be directly evaluated
without initial chemical isolation for clinical efficacy,
(2) increased solubility=bioavailability in crude forms,
and (3) synergy between different active constituents. The
biologically active components of plants useful in the treatment of diabetes include flavanoids, alkaloids, glycosides,
polysaccharides, and peptidoglycans (Grover et al., 2002).
Myristica fragrans Houtt. (Myristicaceae) (MF) is an
evergreen aromatic tree commonly known as nutmeg
or mace, and has been used traditionally as a spice and
for medicinal purposes as carminative, digestive, and
expectorant in the Indian system of medicine. In the recent
Accepted: August 29, 2006
Address correspondence to: D.K. Arulmozhi, Ph.D., Department of Pharmacology, New Chemical Entity Research, Lupin Research
Park, Village Nande, Taluk Mulshi, Pune 411 042, Maharashtra, India. Tel.: þ 91 20 2512 6161; Fax: þ 91 20 2512 6298;
E-mail: adk_bits@yahoo.com
DOI: 10.1080/13880200601028339 # 2007 Informa Healthcare
Antidiabetic effects of Myristica fragrans
literature, MF has been investigated for its hypolipidemic,
antithrombotic, antiplatelet aggregating, antifungal,
aphrodisiac, anxiogenic, anti-ulcerogenic, antitumor,
and anti-inflammatory activities (Ozaki et al., 1989; Park
et al., 1998; Capasso et al., 2000; Sonavane et al., 2001,
2002; Morita et al., 2003; Chung et al., 2006). MF has been
reported to contain 25–30% fixed oils and 5–15% volatile
oils such as camphene, elemicin, eugenol, isoelemicin, isoeugenol, methoxyeugenol, pinen, sabinene, safrol, and
also chemical substances such as dihydroguaiaretic acid,
myristic acid, myristicin, and ligan (Forrest & Heacock,
1972; Isogai et al., 1973; Janssen et al., 1990).
In some preliminary studies, MF was found to have a
hypolipidemic effect in rabbits (Sharma et al., 1995;
Ram et al., 1996). However, no reports of detailed investigations of MF are available on the antidiabetic and
antihyperlipidemic effects. Hence, the objectives of the
current study are to investigate the antidiabetic effect
of MF in mice and its antihyperlipidemic effect in highcholesterol-fed rats.
Materials and Methods
65
National Toxicology Center, Pune, India. On arrival, the
animals were placed at random and allocated to treatment
groups in polypropylene cages with paddy husk as bedding.
Animals were housed at a temperature of 24 2C and
relative humidity of 30–70%. A 12:12 light:dark cycle
was followed. All animals had free access to water and standard pellet laboratory animal diet. All the experimental
procedures and protocols used in this study were reviewed
and approved by the Institutional Animal Care and Use
Committee of Poona College of Pharmacy, Pune, India.
Effect of MF on chlorpromazine-induced hyperglycemia
and hypertriglyceridemia
Male Swiss albino mice were used at 8 weeks of age. Mice
were administered chlorpromazine (10 mg=kg, p.o.), concomitantly with either MF (50, 150, or 450 mg=kg, p.o.) or
glimepiride (10 mg=kg, p.o.) or rosiglitazone (10 mg=kg,
p.o.) or fenofibrate (100 mg=kg, p.o.) for 7 days. Animals
in the control group received vehicle (0.3% Tween 80 in
water, 10 ml=kg, p.o.). Blood samples were collected in
fed state from the animals under mild ether anesthesia from
retro-orbital sinus 1 h after drug administration.
Plant material and extraction procedure
Effect of MF on hyperlipidemic rats
Pharmacognostically identified fruits of Myristica
fragrans Houtt. (Myristicaceae) were collected from the
local market and authenticated by Dr. A.M. Mujumdar,
taxonomist, Agharkar Research Institute (Pune, India).
A voucher specimen (no. M5621) was deposited in the
herbarium of the institute. The hydroalcoholic extract
was prepared as follows: dried and powdered rhizome
(100 g) was soaked in 400 mL of 50% ethanol for 16 h
at room temperature. The percolate was then decanted,
centrifuged, and filtered through Whatman (no. 1) filter
paper to obtain a clear extract. The extracted plant
material was again subjected to the same extraction procedure. The percolates were pooled, concentrated, and
lyophilized (15–18% yield).
Male Wistar rats were made hyperlipidemic by feeding a
high-fat diet containing 2% cholesterol and 1% sodium
cholate mixed with standard laboratory chow
(Chakrabarti et al., 2004) and treated orally with MF
(50, 150, or 450 mg=kg) or fenofibrate for 7 days. Effect
of MF on hepatic triglyceride output was measured by
intraperitoneal injection of tyloxapol at 300 mg=kg
(5 mL=kg in saline). Plasma triglyceride levels were measured at 0, 4, 6, and 24 h after injection.
Chemicals
Chlorpromazine (CPZ), cholesterol, sodium cholate,
tyloxapol, and fenofibrate were obtained from Sigma
Chemical Co. (St. Louis, MO, USA). Rosiglitazone and
glimepiride were gifts from Sun Pharma (Vadodara,
India) and Torrent Pharmaceuticals (Ahmedabad, India),
respectively. All the drugs were solubilized in 0.3% Tween
80 and administered orally between 1000 and 1,100 h for 7
days. All other chemicals and reagents were of pure analytical grade obtained from local suppliers.
Animals
Adult male Swiss albino mice (22–26 g) (six animals per
group per treatment) and male Wistar rats (four to six
animals per group per treatment) were obtained from the
Determination of plasma metabolic parameters
Plasma obtained from the mice and rats was used to estimate the metabolic parameters. Glucose, triglyceride,
and total cholesterol levels were measured spectrophotometrically using commercially available kits (Bayer
Diagnostics, Vadodara, India).
Statistical analysis
Values are expressed as mean values SEM. The statistical significance of differences between the mean values
was analyzed by ANOVA and Dunnett’s test. A p value
of < 0.05 was considered to be significant.
Results
Effect of MF on chlorpromazine-induced hyperglycemia
and hypertriglyceridemia
Administration
of
chlorpromazine
significantly
(p < 0.05) elevated the plasma triglyceride and glucose
66
D.K. Arulmozhi et al.
levels in Swiss albino mice. The triglyceride (TG) values
were found to be 79.33 6.55 mg=dL in vehicle-treated
animals and 125.00 13.15 mg=dL in chlorpromazinetreated animals. Treatment with MF dose-dependently
reduced the elevated TG levels in chlorpromazine
(CPZ)-treated animals and at doses of 150 mg and
450 mg, the effect (81.88 6.93 and 58.78 10.56,
respectively) was statistically significant (p < 0.01) and
comparable with that of the standard drugs glimepiride,
rosiglitazone, and fenofibrate.
Treatment of MF also significantly reduced the elevated glucose levels in CPZ-treated animals. At doses of
150 and 450 mg, the effect was significantly different
from the vehicle-treated animals (246.57 26.17 mg=dL
in vehicle vs. 175.00 6.28 and 143.72 6.16 for
150 mg and 450 mg, respectively). Interestingly, the glucose values were significantly reduced only by rosiglitazone, where glimepiride and fenofibrate had little and
no effect, respectively.
Effect of MF on hyperlipidemic rats
In high-cholesterol-fed rats, MF exhibited dosedependent reduction of triglyceride and cholesterol levels
with respect to vehicle-treated animals. A significant
(p < 0.01) maximum reduction of 47% at 450 mg=kg,
on TG levels, while 66.7% reduction (p < 0.01) was
observed for cholesterol with the same dose. The positive
control, fenofibrate, also significantly (p < 0.01) reduced
TG and cholesterol levels at 100 mg=kg (Table 1).
The low and intermediate doses of MF (50 and
150 mg=kg) did not produce any significant effect on
hepatic triglyceride secretion in high-cholesterol-fed animals, though a mild reduction in TG levels was observed
than in the vehicle animals. However, at 450 mg=kg, MF
significantly (p < 0.05) reduced the TG levels at 24 h after
tyloxapol administration. The positive control, fenofibrate, also reduced the TG levels at various time points
(Figure 3).
Figure 1. Effect of Myristica fragrans on chlorpromazineinduced elevations of plasma glucose levels in mice. Compounds
were administered orally for 7 days. Bars represent means SEM from n ¼ 6. p < 0.005 and p < 0.001 compared with
vehicle treatment. MF, Myristica fragrans; Fenofib, fenofibrate;
Rosi, rosiglitazone; CPZ, chlorpromazine; Glim, glimepiride.
#
p < 0.05 compared to vehicle=vehicle treated animals.
TG concentrations in chlorpromazine-induced type 2
diabetes in mice. This effort was successful, as attested
to by the results shown in Figures 1 and 2, documenting
the fact that MF is capable of significantly lowering
plasma glucose concentrations when given orally to a
mouse model of type 2 diabetes.
Three major classes of oral antidiabetic agents that are
in use today act through three distinct mechanisms:
Discussion
To our knowledge, there is no previous ethnomedical evidence that extracts of Myrsitica fragrans would be useful
in the treatment of hyperglycemia. Though reports on
hypolipidemic effects are available, only a single dose
of MF has been employed in rabbits (Sharma et al.,
1995; Ram et al., 1996). Our decision to explore this
possibility was based on the observation that terpenoidtype quinines isolated from the tree Pycnanthus
angolensis Welw (Myristicaceae), also known as African
nutmeg, have marked antidiabetic effects in mouse models of type 2 diabetes (Luo et al., 1999).
Based on this observation, we have evaluated the ability of crude extract of MF to lower plasma glucose and
Figure 2. Effect of Myristica fragrans on chlorpromazineinduced elevations of plasma triglyceride levels in mice. Compounds were administered orally for 7 days. Bars represent means SEM from n ¼ 6. p < 0.005 and p < 0.001 compared with
vehicle treatment. MF, Myristica fragrans; Fenofib, fenofibrate;
Rosi, rosiglitazone; CPZ, chlorpromazine; Glim, glimepiride.
#
p < 0.05 compared to vehicle=vehicle treated animals.
Antidiabetic effects of Myristica fragrans
67
Table 1. Effect of MF on metabolic parameters in high-cholesterol-fed rats.
Treatment
Vehicle
MF
Fenofibrate
Dose
Triglycerides (mg=dL)
Cholesterol (mg=dL)
10 mL=kg
50 mg=kg
150 mg=kg
450 mg=kg
100 mg=kg
141.82 23.24
154.63 45.20
109.32 13.49
75.56 8.34
64.01 11.70
294.31 29.83
217.94 19.52
190.54 4.44
98.25 18.16
178.35 11.00
Compounds are administered orally for 7 days. Values are expressed as mean SEM from n ¼ 5 to 6, p < 0.005 and
p < 0.001 compared with vehicle treatment. MF, Myristica fragrans.
sulfonylureas by inducing insulin secretion, acarbose by
inhibiting carbohydrate absorption, and thiazolidinediones by insulin sensitization.
It has been reported that chlorpromazine-induces
hyperglycemia in mice (Dwyer & Donohoe, 2003). The
antagonism at muscarinic acetylcholine receptors is suggested as one of the mechanism for the metabolic abnormalities with chlorpromazine (Xie & Lautt, 1995). MF
has recently been reported to possess procholinergic
activity (Parle et al., 2004) and significantly reduced the
carbachol-induced gastric acid secretion in preclinical
models (Jan et al., 2005). These results suggest that the
procholinergic effect may slow the digestion of food
and decrease the carbohydrate absorption. In the current
study, MF significantly reduced the plasma glucose levels
after 7 days of treatment. The effect observed at
450 mg=kg dose was comparable with that of known
thizolidinedione rosiglitazone (10 mg=kg). Also, the
elevated TG levels observed after the administration of
chlorpromazine was significantly lowered with MF
treatment. Because the chlorpromazine-induced metabolic abnormalities (elevated triglycerides and glucose)
have not been studied for the evaluation of antidiabetic
compounds, we have employed glimepiride (a sulfonylurea with extrapancreatic effects), rosiglitazone, a
thizolidinedione, and fenofibrate, a known lipid-lowering
agent, to validate the current model. As expected, all the
three agents significantly reduced the plasma TG levels
after 7 days of treatment, and fenofibrate did not exhibit
any effect on plasma glucose levels.
Male Wistar rats, when fed with a high-cholesterolcontaining diet, showed a significant increase in plasma
total and LDL-cholesterol and also plasma triglyceride
levels with a concomitant decrease in HDL-cholesterol
levels. No change in plasma glucose level was observed.
When treated with MF, these animals showed a dosedependent improvement in plasma lipid levels. The high
cholesterol diet fed rat model has been used previously
to study the efficacy of fibrates (Petit et al., 1988;
Chakrabarti et al., 2004). These animals are hypercholesterolemic, hypertriglyceridemic, but are nondiabetic.
Normally, rodent plasma total cholesterol contains a
very high proportion of HDL-cholesterol and very low
LDL-cholesterol. This makes therapeutic interpretation
of cholesterol lowering in normal rodents difficult. In
the case of hyperlipidemic rats, plasma cholesterol is predominantly LDL-cholesterol, which more closely reflects
the clinical situation. MF showed comparable efficacy at
doses of 150 and 450 mg=kg with fenofibrate (100 mg=kg)
in this model.
MF, at 450 mg=kg, significantly inhibited the lipoprotein secretion in high-cholesterol-fed rats. This assay is
based on intraperitoneally injected tyloxapol to prevent
the catabolism of TG-rich lipoproteins (Li et al., 1996).
Thus, after the administration of tyloxapol, there is a linear increase in the TG levels, which reflects the TG
secretion rate from the liver. It may be postulated that
MF has some effect on the liver TG secretion.
Conclusions
Figure 3. Effect of Myristica fragrans on hepatic triglyceride
secretion after the administration of tyloxapol in high cholesterol diet fed rats. Compounds were administered orally for
7 days. Each point represents mean SEM from n ¼ 4 to 6.
p < 0.005 and p < 0.001 compared with vehicle treatment.
MF, Myristica fragrans; Feno, fenofibrate.
In summary, for the first time we have demonstrated that
the ethanol extract of Myristica fragrans fruit ameliorates hyperglycemia and abnormal lipid metabolism in
animal models. Further pharmacodynamic investigations
in appropriate models of type 2 diabetes are required to
understand the precise mechanism of antihyperglycemia
and antihyperlipidemia exhibited by Myristica fragrans.
68
D.K. Arulmozhi et al.
Acknowledgments
We thank Dr. S.S. Kadam and Dr. K.R. Mahadik (Bharati Vidyapeeth Deemed University, Poona College of
Pharmacy, Pune) for their constant encouragement in
the work.
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