INTRODUCTION:
Rheumatoid arthritis is an autoimmune disease in which
there is joint inflammation, synovial proliferation and destruction of articular cartilage.1
It is a common disease having peak incidence in 3rd
to 4th decades of life with 3-5 times higher preponderance in
female.2 Its prevalence
depends upon age.3 Herbal drugs constitute a
major part in all the traditional system of medicine. Herbal medicine is a
triumph of popular therapeutic diversity.4
The factors responsible for the continued and extensive use of herbal remedies
in India are their effectiveness, easy availability, low cost, comparatively
less toxic effects and shortage of practitioners of modern medicine in rural
areas.5 Number of synthetic medicines has
been derived from medicinal herbs.6
The sapodilla is large, evergreen, forest tree more than 30 m in height and with
a diameter up to 1.5 m, under cultivation it varies between 9 and 15 m,
depending on location, and generally does not exceed 50 cm in diameter. The
gummy latex of sapodilla, called chicle, is used to
make chewing gums, and the fruit is used to treat diarrhea and pulmonary
diseases. 7 The crushed seeds are claimed to expel bladder and kidney
stones and effective in rheumatism.8
Leaf decoction used for fever, hemorrhage, wounds and ulcers. 9 Hence, the present
study was undertaken to evaluate in vitro
antiarthritic activity of plant extract.
MATERIALS AND
METHODS:
Plant material:
The leaves of plant Manilkara
zapota were collected from local area of Bhopal,
M.P., India, and
authenticated by Mr Anil Prakash,
department of Biotechnology, Barkatullah university,
Bhopal, M.P. where a voucher specimen No. has been submitted. (Voucher specimen
No. 28 SAP)
Preparation of plant extract:
Collected leaves of Manilkara
zapota were converted into moderately coarse
powder and extracted with solvent ethyl alcohol for 27 hours by soxhlet. The solvent was removed under reduced pressure and
the yield of extract was calculated.
ASSESSMENT OF IN VITRO ANTI-ARTHRITIC ACTIVITY:
Inhibition of protein denaturation
method:10
The reaction mixture (0.5 ml) consisted of
0.45 ml bovine serum albumin (5% aqueous solution) and 0.05 ml of Manilkara zapota extract (100 and 250 mcg/ml of final volume). pH was adjusted at 6.3 using a small amount of 1 N HCl. The samples were incubated at 37˚C for 20 min and
then heated at 57˚C for 30 min. After cooling the samples, 2.5 ml
phosphate buffer saline (pH 6.3) was added to each tube. Turbidity was measured
spectrophotometrically at 660 nm for control test
0.05 ml distilled water was used instead of extracts while product control test
lacked bovine serum albumin. The percentage inhibition of protein denaturation was calculated as follows.
Percent inhibition=
100- (O.D. of test – O.D. of product
control) X 100
O D.. of control
The control represents 100% protein denaturation. The results were compared with acetyl
salicylic acid (250 mcg/ml) treated samples.
Statistical
Analysis:
Data are presented as the mean ± SEM of
each triplicate test. The analysis was performed by using Dunnett
vs. Control test and by ANOVA. P‘0.05 were considered to be statistically
significant.
RESULTS AND
DISCUSSION:
Anti-arthritic effect of ethanolic extract of Manilkara
zapota was studied significantly by using in-vitro
inhibition of protein denaturation model. The effect
of ethanolic extract of Manilkara zapota
on inhibition of protein denaturation is shown in
table 1. Extract of Manilkara
zapota at two different concentrations (dose
levels) provided significant protection against denaturation
of proteins. Most of the investigators have reported that denaturation
of protein is one of the cause of rheumatoid arthritis. Production of
auto-antigens in certain rheumatic diseases may be due to in vivo denaturation of proteins.
Mechanism of denaturation probably involves alteration
in electrostatic, hydrogen, hydrophobic and disulphide bonding. Obtained data
stated that Manilkara could be used as
potent anti-arthritic agent.
Table 1 Effect of ethanolic extract of Manilkara zapota on
inhibition of protein denaturation
|
S. N. |
Groups |
Protein denaturation (%) |
|
1 |
Manilkara zapota (100 mcg/ml) |
58.89 ± 6.29 |
|
2 |
Manilkara zapota (250 mcg/ml) |
75.84 ± 2.31 |
|
3 |
Acetyl salicylic
acid |
51.25 ± 7.14 |
Values (Mean ± SD)
are presented in comparison to control and standard.
CONCLUSION:
From the results obtained in the present
study, it may be concluded that Manilkara zapota possess significant anti-arthritic activity.
Hence it could be beneficial for further work as active anti-arthritic agent.
ACKNOWLEDGEMENT:
Mr. Yogesh Shivhare acknowledged to Mr. Rakesh
Kumar Punekar vice-principal, RKDF college of
Pharmacy, for their kind support during the progress of this work.
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Received on 22.10.2011 Accepted on 28.11.2011
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Asian J. Pharm. Tech. 1(4): Oct. - Dec. 2011; Page 123-124