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International Journal of Toxicological and Pharmacological Research 2017; 9(1); 56-63
ISSN: 0975-5160
Research Article
*Author for Correspondence: arifur.mpharm@gmail.com
Phytochemical and Pharmacological Activity of Myristica fragrans
Houtt (Myristicaceae)
Md Arifur Rahman Chowdhury1,2*, Manirujjaman1, 3, Md Mazedul Haq4
1Department of Pharmacy, Southeast University, Banani, Dhaka, Bangladesh.
2Department of Pharmacy, State University of Bangladesh, Dhaka, Bangladesh.
3Faculty of Health, School of Clinical Sciences, Queensland University of Technology, Brisbane, Australia.
4 Department of Biochemistry, National University, Gazipur, Bangladesh.
Available Online: 1st March, 2017
ABSTRACT
Objective: Myristica fragrans Houtt commonly used as traditional medicine for alleviating of various disorders. The
purpose of our study was to map out the in vitro antioxidant property and in vivo anti-hyperglycemic and analgesic effect
of the methanolic extract of Myristica fragrans Houtt. (Seed and mace) (Myristicaceae) on Swiss albino mice. Methods:
The processed powder of Myristica fragrans Houtt (seed and mace) were subjected to methanolic extraction by soxhlet
filtration methods, and the desiccated extract was used for screening of antioxidant by DPPH free radical scavenging
assessment as well as total phenolic content by using folin-ciocalteu reagent.Anti-hyperglycemic effect and analgesic
action tested through alloxan induced antidiabetics test and acetic acid-tempted writhing test on mice. Results: In DPPH
free radical scavenging assessment, free radicals neutralization expressed as % of inhibition 49.69±0.06% also by IC50
values as 68.43 µg/ml surmise middle level of antioxidant property. The total phenolic content expressed as 186.25 mg/g
equivalent of gallic acid indicates, active phenolic content. Oral administration of 200 and 400 mg/kg of extract dose and
reference drug vildagliptin (50 mg/kg) for the duration of the 4-day study period, and initiated % of inhibition the blood
glucose level measured as 22.48%, 44.78% and 62.02% regard as the significant anti-hyperglycemic properties. The
analgesic activity was investigated by using the acetic acid-induced writhing test in mice, at the dose of 200 mg/kg body
and 400mg/kg weight, and resulting 50.4% and 68.10% correspondingly, which was considerably significant with a
standard drug. Conclusion: The present study suggests that methanolic extract of seed and mace of Myristica fragrans Houtt
can manage moderate oxidative stress as well as perform the painkilling action. Besides, prolong medication may enhance
the new dimension of anti-hyperglycemic activity.
Keywords: Antioxidant, Total Phenolic content, Myristica fragrans Houtt as (Mfsm), Analgesic activity, Writhing test,
Anti-hyperglycemic test.
INTRODUCTION
Which components can promote health and alleviate
illness manifested as “Medicine” and medicine belongs to
chemical compounds. The large diversified chemical
compounds like phytochemicals (alkaloids, flavonoids,
tannins, and phenolic) constituents are present in plants,
which are biologically active to show pharmacological
effects. Such chemical substances can act as an originator
for synthesis drugs.For reducing demoralizing illness with
significant morbidity and mortality, the medicinal plant
considered as a safe remedy (cardiovascular problems,
cancer, liver disorders, central nervous system, digestive
and metabolic disorders) and chemopreventive actions and
costs effectiveness, less toxic1,2,3-8,9.
In this phytochemical and pharmacological assay,
Myristica fragrans Houtt (Mfsm) (seed and mace) tested for
their oxidative stress inhibitory effect, analgesic, and anti-
hyperglycemic effect. Myristica fragrans Houtt.(Nutmeg,
mace –English; Jai phal –India; Seed and mace- Jai phal-
Jaiyatri Bangladesh; Pala -Indonesian) Belonging to the
family of Myristicaceae is a scented evergreen tree that
grows 30–39 ft (1 ft=30.48 cm) high with spreading
branches and yellow fleshy fruits, having an appearance
like apricot or peach. Both the seed (Nutmeg) and its fleshy
aril (mace) are known as spices10. From the ancient time,
it used as a traditional medicine for the remedial purpose
of a broad range of disease. Nutmeg useful for curing
Diarrhea, mouth sore and insomnia and inhibitory activity
against several kinds of anaerobic and aerobic
microorganisms11. Besides,Myristica fragrans still used
as, antioxidant, analgesics, stomachic’s, aphrodisiacs,
digestives, hypnotics; amenorrheal agents12,13. The
significant reducing capability of bacterial infection14 and
as an aphrodisiac, anti-rheumatoid, anti-malarial,
stimulant, and post childbirth15 also reported.From the
experimental evidence, Myristica fragrans Houtt exhibit
potent antifungal and antibacterial (the majority of gram
positive and gram negative) activities16,17-19. At the same
time, Myristica fragrans Houtt used as food
preservatives20, antiseptic and disinfectant21. In another
Md. Arifur et al. / Phytochemical and Pharmacological…
IJTPR, Volume 9, Issue 1, February- March 2017 Page 57
study, confirmed that nutmeg extract had significant
prevention capability of hypercholesterolemia and
atherosclerosis22 and hepatoprotective activity23. Even it
has proven high anti-inflammatory action24,25-27 and
antidepressant effect28.
Thus, we studied the methanolic extract of Myristica
fragrans Houtt(Mfsm) seed and mace for it antioxidant
property, along with analgesic and antidiabetic effects.
MATERIALS AND METHODS
Chemicals and solvents
DPPH (2, 2-Diphenyl-1-picrylhydrazyl) was obtained
from Sigma Chemical Co., USA. Methanol, ascorbic
acid,folin-acid, folin-ciocalteu/folins phenol reagent,
sodium carbonate (Na2CO3), gallic acid, alloxan from
(Fluka, Germany), vildagliptin-50mg/tablet collected from
drug manufacturer - Novartis, Bangladesh; and glucose
estimation kit (Human, Germany). All the chemicals used,
including the solvents, were of analytical grades.
Instruments
HACH DR 4000U UV-visible spectrophotometer
equipped with quartz cells of the 1-cm light path used to
determine the molecular absorption spectra and
absorbance at specific wavelengths.
Collection of the plant parts
The fresh, dried seed and mace of Myristica fragrans Houtt
(Mfsm) were collected from a local savory spice shop,
Mirpur, Dhaka, as well as from Sylhet district, Bangladesh.
Shapna Sultana, Lecturer, Department of Pharmacy,
Southeast University, Banani, primarily identified the
collected, dried seed and mace. As a final point of
attribution by a taxonomist at "Bangladesh National
Herbarium" Mirpur, Dhaka, Bangladesh a receipt
specimen was deposited in the herbarium division
numbered as DACB (Accession Number 43646).
Drying, pulverization, preservation of plant parts
The collected seed and mace were washed with fresh water
and dried for one week. The seed and mace collectively
grounded into a coarse powder with the help of a suitable
grinder machine and passing through a sieve, in
pharmaceutical technology lab, Southeast University,
Dhaka. As a final point, crushed powder stored in a cool,
dark, and dry place in an airtight container, and kept until
analysis commenced.
Extraction of the plant material
By using soxhlet apparatus dried plant material were
refluxed with methanol. The vapor flows through a coil
where they condense back to a liquid which is then
collected in the receiving vessel. The whole mixture was
successively filtered through a piece of clean, white cotton
material and Whatman filter paper (Bibby RE200, Sterilin
Ltd., UK).
Experimental animals
The Swiss albino mice of both sexes weighing (29.3±3.9)
were used to conduct the research and procured from the
animal research branch of the (icddr,b-International Centre
for Diarrhoeal Disease and Research) Bangladesh. They
kept under standard husbandry conditions (temperature
23±2 C relative humidity 55± 10 and 12 light and 12-hour
dark cycle).The animals fed with commercial diet pellets
and water ad libitum. Before experimentation session, the
animals were allowed to acclimatize to the atmosphere for
seven days. Animals were kept fasting overnight but
allowed free access to water.
Ethical approval
As per University requisition, icddr,b; provided the
animals for experimenting purpose. All the testing on
animals were performed by guidelines of the institutional
animal ethics committee29 Southeast University, Banani
Dhaka, Bangladesh.
In vivo test
Antioxidant activity measured by DPPH (1,1-diphenyl-2-
picrylhydrazyl) free radical scavenging assay
In a chemical reaction with dpph, antioxidants stabilized
the free radical and reduced to dpph-H. As a consequence;
the absorbance has decreased from the DPPH radical to the
DPPH-H form. The scale of yellowing indicates the
scavenging potential of the antioxidant (Hydrogen
donating capacity) compounds30. The scavenging reaction
between (DPPH) and plant extract [an antioxidant (H-A)]
can be expressed as:
(DPPH) + (H-A) -----------------------DPPH-H + (A)
(Purple) (Extract) (Yellow)
The free-radical scavenging activity of Myristica fragrans
Houtt (Mfsm) extracts was measured by a decrease in the
absorbance of a methanol solution of DPPH31. A stock
solution of DPPH (400 μg/ml) was prepared in methanol,
and 100 μl of this stock solution was added to 5 ml of
solutions of Myristica fragrans Houtt (Mfsm) extracts of
different concentrations (5, 10, 25 and 50 μg/ml)32. The
solutions were then mixed properly and kept in the dark for
20 min, and the absorbances were measured at 517 nm by
using a HACH DR 4000U UV-visible spectrophotometer.
Scavenging activity was expressed as the percentage
inhibition calculated using the following formula as, the
percentage of inhibition = [(A0 – A1) / A0] x 100; Where,
A0 = Absorbance of the control; A1 = Absorbance of the
plant extract/ standard. And the IC50 value was calculated
from the equation of line obtained by plotting a graph of
concentration (µg/ml) versus % inhibition33. Ascorbic acid
used as a reference standard. The radical scavenging
activity (RSA) or percentage inhibition evaluated by
comparing the values of absorbance for the investigational
samples and control following the equation as indicated34.
Total phenolic content test
By using folin-ciocalteu reagent (FCR - a hetero-poly
phosphotungstates-molybdates utilized for the
colorimetric assay of phenolic and polyphenolic
antioxidants), the total phenolic content of certain plant
materials can be measured, which help to measure
reducing capacity. Sequences of reversible one or two
electron reduction reactions lead to blue species possibly
(PMoW11 O40)4. In essences, it is assumed that the
molybdenum is easier to be reduced in the complex and
electron transfer reaction occurs between reluctant and Mo
(VI). The overall procedure performs by using folin-
ciocalteu reagent35 and aluminum chloride method36,37
correspondingly. The content of phenolic in the extract of
Md. Arifur et al. / Phytochemical and Pharmacological…
IJTPR, Volume 9, Issue 1, February- March 2017 Page 58
Myristica fragrans Houtt (Mfsm) was calculated from
regression equation of the calibration curve and expressed
as gallic acid standard /equivalent (GAE)38.
In vivo test
Analgesic activity (Acetic acid-tempted writhing in mice):
The analgesic activity of Myristica fragrans Houtt (Mfsm)
assayed by the acetic acid induced writhing test39. One
hour before peritoneal insertion of acetic acid 300 mg/kg
(3% solution in sterile distilled water) i.p, diclofenac-Na
used as reference standard drug and Myristica fragrans
Houtt (Mfsm) extract at different concentration
administrated orally. After 20 minutes of addition of acetic
acid, the writhing of each counted and the percent of
inhibition was calculated by comparing with control group.
If the sample possesses analgesic activity, it will
subordinate the number of writhing than the standard and
revealed the analgesic activity as % inhibit of writhing.
Anti-hyperglycemic activity on alloxan induced diabetic
mice
To complete the antidiabetic activity, indiscriminately
separated group of mice was injected intraperitoneally
(I.P.) at a dose of 120 mg/kg b.w. Freshly prepared alloxan
monohydrate (fluka, Germany). After an hour of alloxan
administration, animals were fed ad libitum and 1ml of
(100 mg/ml) glucose i.p. to combat ensuring severe
hypoglycemia after 72 hr of alloxan injection; the animals
were tested for evidence of diabetes by estimating their
blood glucose level using a glucometer. Mice showed FBG
> 150 mg/dl considered diabetic and selected for studies.
Animal grouping and experimental design
Animals selected fasted overnight and then divided into
four groups (n=4) as follows
Group-I: Normal control mice (non-alloxanized) that
administered distilled water only; Group-II:diabetic
control mice (untreated, alloxanized); Group-II: Diabetic
mice administered with Myristica fragrans Houtt (Mfsm)
(200 mg/kg/day) respectively; Group-IV: Diabetic mice
administered with Myristica fragrans Houtt (Mfsm) (400
mg/kg/day) respectively; Group-V: mice administered
once with vildagliptin (50 mg /kg) as reference standard
drug.
Statistical Analysis
All the tests were performed triplicates and values are
expressed as Mean ± Standard Deviation /SEM by using
Microsoft Excel, 2007.
RESULTS
DPPH free radical scavenging activity
Methanolic extract of Myristica fragrans Houtt (Mfsm)
demonstrated proton /H-donor activity and showed potent
scavenging rate of dpph. The percentage (%) of
scavenging was found to be concentration dependent, i.e.,
scavenging capacity increases with the increase of the
concentration of both the extracts. By contrasting % of
inhibition for (Mfsm and AA for the concentration of 80
µg/ml, found 49.69±0.06% and 88.22 ±0.10% (Table: 1
and Table: 2) respectively, which indicate the median
antioxidant capacity of sample Myristica fragrans Houtt
(Mfsm). Additionally, the IC50 values of (Mfsm) and AA is
68.43 µg/ml. and 12.82 µg/ml calculated by using
regression equation YMfsm = (0.383x +23.79; R2=0.575)
and (YAA =0.659x+41.55; R2=0.777) respectively.
Furthermore, the % of inhibition depicted in Fig:1 reveals
the insignificant difference of Mfsm and AA.
Determination of total phenolic content
The total phenolic content in the extracts of Myristica
fragrans Houtt (Mfsm) determined according to the
colorimetric folin-ciocalteu assay with gallic acid as a
standard compound. The total phenolic content was
calculated from regression equation curve
(y=0.012x+0.342, R2=0.929) and expressed as gallic acid
equivalent (GAE) (Table:3 and Table:4 ).A potent range of
total phenolic content was found in the plant materials to
be186.25 mg/g plant extract (in GAE-Table 3) for
Myristica fragrans Houtt (Mfsm) (Table 4).
Calculation
Regression equation y=0.012x+0.342, R2=0.929; y=0.789;
m=0.012; c=0.342
x= 37.25 micro gm/ml; C= 0.0373 mg/ml;
V=1 ml; m=.0002
A = (C × V)/m
= 186.25 mg/g Gallic Acid
Where, A = Total phenol content, mg/g plant extract,
in Gallic acid
C= the concentration of Gallic acid established from
the calibration curve, mg/ml
V = the volume of extract, ml; m = the weight of
pure plant methanolic extract, g
The amount of total phenol content of Myristica fragrans
Houtt (Mfsm) is 186.25 mg/g gallic acid (GA) Acid
Analgesic activity
Animal were divided into four groups (4 animals in each
group), as group 1: Control (Saline water treated); group
2: Positive control (Diclofenac treated);group 3: Myristica
fragrans Houtt (Mfsm) 200mg/kg; group 4: Myristica
fragrans Houtt (Mfsm) 400mg/kg.After administration of
the different dose of methanolic extract of Myristica
fragrans Houtt (Mfsm) and standard drug diclofenac as the
positive control, the numbers of writhing reduced and
ensured analgesic effects. By comparing the number of
writhing with the untreated control group Myristica
fragrans Houtt (Mfsm) showed almost similar % of
inhibition as 68.10% at the dose of 400mg/kg. (Table 5)
Antidiabetic activity on alloxan-induced diabetic mice
The consequences after chronic administration of
Myristica fragrans Houtt (Mfsm) showed meaningful anti-
hyperglycemic action between investigational and diabetic
control mice. At a dose of 200 mg/kg body weight,
Myristica fragrans Houtt (Mfsm) significantly lowered
blood glucose level and showed reduction of 22.48 %
while at 400 mg/kg Myristica fragrans Houtt (Mfsm) body
weight dose, produced maximum reduction of 44.78% of
blood glucose level, respectively, inhibition of blood
glucose level 62.01 % was found for vildagliptin (50 mg
/kg) on day 4 as a peak. (Table 6).
DISCUSSION
In vitro antioxidant activity
In vitro findings indicated that Myristica fragrans Houtt
(Mfsm) recognized their anti-oxidative abilities regarding
Md. Arifur et al. / Phytochemical and Pharmacological…
IJTPR, Volume 9, Issue 1, February- March 2017 Page 59
Table 3: Absorbance of gallic acid at different
concentration.
Conc.
Abs
Abs
Abs
Avg
200
2.890
2.870
2.930
2.897
150
2.110
2.190
2.170
2.157
100
1.390
1.420
1.450
1.420
50
1.021
1.040
0.990
1.017
Note: All absorbance presented as Abs and
concentration as Conc.
DPPH radical scavenging activity and total phenolic
content.
DPPH radical scavenging activity
Myristica fragrans Houtt traditionally used as medicinal
plant over the decades. Many experimental studies have
reported that the methanolic/acetone extract of nutmeg
seed showed good antioxidant activity by methods of 1,1-
diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing
antioxidant power (FRAP) due to high content of
tannin,flavonoids and terpenoids40,41-44. The first objective
of this work was to evaluate the phytochemicals
composition on the Mfsm .In the present study, the Mfsm
confirmed good antioxidant properties as IC50 values,
contrast with AA is 12.82 µg/ml and 68.43 µg/ml
correspondingly. The obtained result demonstrated that the
Mfsm showed an important ability to donate a proton or
neutralizing capacity of free radicles such as DPPH
radicles with % of inhibition 49.69±0.06% compared with
AA (Ascorbic acid) 88.22 ± 0.10% at a concentration of 80
µg/ml.
According to the experimental assay of Srinivasanet al.,
2005& Gupta et al. 2013 [45-46] spices possession as
medicinal properties and our finding may anticipate the
Table 1: The percentage of inhibition of methanolic extracts of Myristica fragrans Houtt (Mfsm).
Conc.
Abs 1
Abs 2
Abs 3
% of inhibition
Avg %
SEM
IC50
0
1.775
1.777
1.779
0.000
0.000
0.000
0.00
0.00
68.43
5
1.591
1.588
1.589
10.366
10.636
10.680
10.56
0.10
10
1.207
1.209
1.203
32.000
31.964
32.378
32.11
0.13
20
1.010
1.030
1.070
43.099
42.037
39.854
41.66
0.96
40
0.989
0.991
0.983
44.282
44.232
44.744
44.42
0.16
80
0.891
0.894
0.897
49.803
49.690
49.578
49.69
0.06
Note: All values of absorbance are mean ± SEM of triplicates.IC50 (regression equation calculated the inhibitory
concentration by half or 50%).
Table 2: The percentage of inhibition of ascorbic acid (AA).
Conc.
Abs 1
Abs 2
Abs 3
% of inhibition
AVG%
SEM
IC50
0
1.785
1.810
1.779
0.000
0.000
0.000
0.00
0.00
5
1.281
1.279
1.265
28.235
29.337
28.893
28.82
0.32
10
0.839
0.832
0.827
52.997
54.033
53.513
53.51
0.30
12.82
20
0.669
0.671
0.674
62.521
62.928
62.114
62.52
0.24
40
0.419
0.401
0.418
76.527
77.845
76.504
76.96
0.44
80
0.209
0.211
0.213
88.291
88.343
88.027
88.22
0.10
Note: All values of absorbance are mean±SEM of triplicates.IC50 (regression equation calculated the inhibitory
concentration by half or 50% ).
Figure 1: The antioxidant activity of methanolic extracts of Myristica fragrans Houtt (Mfsm) and standard ascorbic
acid (AA) determined by using DPPH method.
Table 4: Absorbance of methanolic extract of Myristica
fragrans Houtt (Mfsm).
Total phenolic content
Myristica fragrans Houtt (Mfsm)
Conc.
Abs
Abs
Abs
AVG
ST. DE
0.0002
0.781
0.797
0.789
0.789
0.008
Note: All absorbance presented as Abs and
concentration as Conc.
0.00
20.00
40.00
60.00
80.00
100.00
5
% Inhibition
Concentration (µg/ml)
Ascorbic Acid Myristica fragrans Houtt (Mfsm)
Md. Arifur et al. / Phytochemical and Pharmacological…
IJTPR, Volume 9, Issue 1, February- March 2017 Page 60
ability to care for the consumer’s health from various free
radical-related diseases.
Total phenol content
The second purpose of this study was to evaluate the
phenolic compound by considering Plants are the primary
source of the phenolic compound47. Many studies focused
on the correlation of antioxidant activity to phenolic
compounds content. The results of Kumar et al.48 Yao et
al.49 and Hinnenburg et al.50 stated a strong correlation
between phenol and antioxidant activity.
Proceeding randomized experiments show different types
of phenolic compounds, which a hydroxyl group attached
to the benzene ring and reveal as an active antioxidant51,52.
Through our evaluation, we found the amount of total
phenol content of Myristica fragrans Houtt (Mfsm) is
186.25 /g Gallic Acid, which indicates potent phenolic
content and helps to initiate the anti-hyperglycemic test
and analgesic test.
In vivo anti-diabetic effect
The previous randomized clinical trial demonstrated that
Myristica fragrans Houtt can reduce the blood sugar level
reduced significantly in alloxan induced diabetic rats as
well as increase the insulin secretion53. Also, Myristica
fragrans Houtt (seed) possess a significant dual agonist
PPAR α/γ that ensure the potential anti-diabetic agent for
the treatment of type 2 diabetes54.
In our experiment type -1 (Insulin-dependent), diabetes
study model has been developed to investigate the anti-
hyperglycemic activity of Myristica fragrans Houtt
(Mfsm). By using alloxan (a beta-cytotoxin) insulin-
secreting pancreatic β-cells destructed (a reactive oxygen
species-dependent oxidative damage [55] and resulting in
the diminished level of serum insulin56.
In our study investigation vildagliptin, 50 mg/kg/day as
standard drug and Myristica fragrans Houtt (Mfsm) extract
dose at 200 and 400mg/kg administrated and 62.01
%,22.48 % and 44.78%, inhibition the blood glucose level
correspondingly on day four (04) as a peak. Chronic
administration of Mfsm in diabetic mice resulted in an
effective lowering of blood glucose level suggesting that
the extracts might possess insulin-like effect on peripheral
tissues either by promoting glucose uptake and metabolism
or inhibiting hepatic gluconeogenesis since alloxan
treatment causes permanent destruction of β-cells.
Analgesic effect
The writhing test is a very active method for preliminary
evaluation of anti-nociceptive activity and the % inhibition
of writhing values obtained in animals using this test
correlated with the analgesic doses in humans,[57] still it
Table 5: Analgesic activity of methanolic extract of Myristica fragrans Houtt (Mfsm) on mice presented.
Group
Number of writhing
Mean
SD
SEM
Mean ±SEM
% of Inhibition
m1
m2
m3
m4
Control
32
31
29
33
31.25
1.71
0.85
31.25±0.85
00.00
Positive control
8
7
9
7
7.75
0.96
0.48
7.75±0.48
73.28
G -1( Mfsm)200mg/kg
16
14
17
15
15.5
1.29
0.65
15.5±0.65
50.4
G-2 (Mfsm) 400mg/kg
9
10
7
11
9.25
1.71
0.85
9.25±0.85
68.10
Note: All values of writhing are mean±SEM (n=4).The percentage of inhibition of writhing in acetic acid induced mice.
Table 6: Blood glucose level of alloxan-induced diabetic mice after treatment with the methanolic extracts of Myristica
fragrans Houtt (Mfsm) 200mg/kg and 400mg/kg -body weight for consecutive four (04) days.
Normal control
M1
M2
M3
M4
Mean
SD
SEM
% of Inhibition
5.7
5.4
6
5.5
5.65
0.26
0.13
------
5.4
6.1
4.9
5.7
5.525
0.51
0.25
5.2
5.7
5.3
5.5
5.425
0.22
0.11
6.3
5.1
5.8
5.3
5.625
0.54
0.27
Alloxanized
control mice-untreated
13.4
13.9
15.2
14.9
14.35
0.84
0.42
-----
12.6
13.3
13.5
14.7
13.52
0.87
0.44
16.4
17.2
16.7
16.9
16.8
0.34
0.17
15.3
15.7
16.2
16.4
15.9
0.50
0.25
Myristica fragrans Houtt
(Mfsm) 200mg/kg
12.4
10.9
10.4
9.9
10.9
1.08
0.54
20.16
12.9
11.1
10.8
10
11.2
1.22
0.61
22.48
13.7
13.1
12.3
11.2
12.57
1.08
0.54
18.25
13.1
11.8
11.9
10.3
11.77
1.15
0.57
21.37
Myristica fragrans Houtt
(Mfsm) 400mg/kg
12.6
10.6
9.1
7.4
9.93
2.21
1.11
41.27
12.3
9.4
8.9
7.8
9.6
1.92
0.96
36.59
13.4
10.3
9.2
7.4
10.08
2.52
1.26
44.78
12.7
9.8
8.5
7.6
9.65
2.22
1.11
40.16
Vildagliptin50mg/kg
11.3
6.9
6.5
5.2
7.475
2.65
1.33
53.98
12.5
7.2
5.9
5.1
7.675
3.33
1.67
59.2
13.1
7.3
6.4
5.4
8.05
3.54
1.73
58.77
12.9
7.6
6.1
4.9
7.875
3.53
1.76
62.01
Note: The value of glucose level expressed as Mean ± SEM where N=4.Glucose level suppression expressed as a form
of % of inhibition.
Md. Arifur et al. / Phytochemical and Pharmacological…
IJTPR, Volume 9, Issue 1, February- March 2017 Page 61
cannot indicate whether the effects result from central /or
peripheral actions. The analgesic potential of the extract
was shown by acetic acid test to be significant but was not
accurate58.
The acetic acid-induced writhing model represents pain
sensation by triggering localized inflammatory response.
Such pain stimulus leads to the release of free arachidonic
acid from tissue phospholipids59.
The acetic acid-induced writhing response is a painful
procedure to evaluate peripherally acting analgesics. The
response is thought to be mediated by peritoneal mast
cells60 acid-sensing ion channels61 and the prostaglandin
pathways62.
The previous investigation showed that several nutmeg
(Myristica fragrans) preparations are used to relief sprains,
rheumatism and paralysis63 with analgesics agents as
similar to non-steroidal anti-inflammatory drugs64.
In our study, the acetic acid induced writhing reflexes were
used to explicate central and peripheral antinociceptive
effects. The mean number of abdominal constriction after
I.P. injection of acetic acid on mice compared after
treatment with diclofenac sodium (50 mg/kg) and
methanolic extract of Myristica fragrans Houtt (Mfsm) at
the dose of 200mg/kg and 400mg/kg. A dose-dependent
reduction in the number of writhing was observed in
different concentration and depicted as % inhibition of
writhing response as 73.28%, 50.4% and 68.10%
correspondingly.
CONCLUSION
The complete test indicates that Myristica fragrans Houtt
(Mfsm) have antioxidant potential as well as phenolic
content property which help to manage the hyperglycemic
condition by 40-43% and showed useful analgesic activity
in acetic acid induced writhing method. Further studies are
needed to spot out the bioactive structural association with
the anti-hyperglycemic and analgesic activity.
ACKNOWLEDGEMENTS
The authors are grateful to the Department of Pharmacy,
Southeast University Banani, Dhaka, Bangladesh and the
State University of Bangladesh.
COMPETING INTERESTS
The authors have declared that no competing interests
exist.
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