This is a project entitled of Evaluation of Ex-VIVO Anti inflammatory, anti asthmatic activities of Methanolic extract of Mimosa arenosa. This research was conducted under the guidance of Dr. S VIDYADHARA, and Dr. D. Sandeep in CHEBROLU HANUMAIAH INSTITUTE OF PHARMACEUTICAL SCIENCES, Guntur. Andhra Pradesh, India
Evaluation of Ex-VIVO Anti inflammatory, anti asthmaticactivitiesof methanolic extract of Mimosa arenosa.pptx
1. EVALUATION OF EX-VIVO ANTI-INFLAMMATORY AND
ANTI-ASTHMATIC ACTIVITIES OF METHANOLIC EXTRACT
OF Mimosa arenosa
K. V. Surya Siva Kumar*, D. Sandeep, S. Vidyadhara
Chebrolu Hanumaiah Institute of Pharmaceutical
Sciences, Chandramoulipuram, Chowdavaram,
Guntur, Andhra Pradesh - 522019
2. The key features of inflammation are redness, warmth,
swelling and pain.
Causes:
Physical – Burns, Injuries, Dirt.
Chemical – Irritants, Alcohol.
Biological – Pathogens, Hypersensitivity, Stress.
Treatment:
Non- specific COX inhibitors: Aspirin, Ibuprofen, Diclofenac.
Selective COX-2 inhibitors: Celecoxib
Preferential COX-2 inhibitors: Nimesulide
INFLAMMATION
3. Key features – Breathlessness, Mucosal edema and
Increased secretions.
Causes:
Trigger factors - Infection, Irritants, Pollution, Exercise,
Exposure to cold air, Psychogenic status etc.
Allergic mediators – Histamine, PAF and Leukotrienes.
Mast cells and Allergic antibodies (IgE)
Treatment:
Brochodilators. Eg: Salbutamol, Terbutaline.
Anti-cholinergics. Eg: Ipratropium, Atropine.
Anti-histaminics. Eg: Ketotifen
ASTHMA
5. MATERIALS AND METHODS
REQUIREMENTS – Indomethacin, Histamine, Acetylcholine
EXPERIMENTAL ANIMALS - Healthy adult male albino rats
weighing
250–300g.
Pharmacological Evaluation methods:
1. Ex-Vivo Anti-inflammatory activity - Heat induced haemolysis
& Protein denaturation
2. Ex-Vivo Anti-asthmatic activity – Antihistaminic model using
chick ileum
Preparation of Extract:
Methanolic extract of Mimosa was prepared by maceration
process.
6. EX-VIVO ANTI-INFLAMMATORY ACTIVITY
Heat induced haemolysis:
20µL of uncoagulated fresh rat blood was added to vials containing 1 mL of
0.1M PBS (pH 7.4).
2ml of extracts were added to the vials so as to achieve the final concentrations
of 100, 150, 200, 250 and 300 µg/ml of each sample.
PBS (1ml) and rat blood was added to control vials.
Then the solutions were subjected to centrifugation at 3000 rpm for 10 minutes.
The contents in vials were incubated at 370C for 15 minutes. Then the mixtures
were heated for 25 minutes at 540C.
The supernatant was measured at 540 nm in a spectrophotometer.
%Inhibition of haemolysis
= Absorbance in control group – Absorbance in test group X 100
Absorbance in control group
7. EX-VIVO ANTI-INFLAMMATORY ACTIVITY
Protein Denaturation:
0.2 mL of egg albumin was added to vials containing 2.5 mL of 0.1 M PBS (pH
6.4).
2 ml of extract is added to the vials, so as to achieve the final concentrations of
100, 150, 200, 250 and 300 µg/ml of each sample.
PBS (2.5 ml) and egg albumin was added to control vials.
Then the mixtures were incubated at 37 ± 20C in a BOD incubator for 15
minutes and then heated at 700C for 5 minutes.
After cooling, their absorbance was measured at 660 nm using vehicle as blank.
%Inhibition of denaturation
= Absorbance in control group – Absorbance in test group X 100
Absorbance in control group
8. EX-VIVO ANTI-ASTHMATIC ACTIVITY
Isolated chick ileum suspended in tyrode solution maintained at 37±0.5°C
Record a dose response curve for histamine at variant molar
concentrations
Then add methanolic extract of Mimosa to reservoir and repeat the
same doses of histamine
Graph of percentage of maximum contractile response on Y-axis and
Log dose of histamine on X-axis was plotted to record dose response
curve of histamine, in absence and in presence of extract
10. Effect of Methanolic extract of Mimosa arenosa on Heat
induced haemolysis of Erythrocyte membrane
S. No
Concentration
(µg/ml)
% Inhibition of Heat induced haemolysis (Mean ± SEM)
SD (Indomethacin) MME
1 100 34.26 ± 1.1222 36.31 ± 1.5015
2 150 44.74 ± 0.6854 48.12± 1.1698
3 200 58.75 ± 1.1847 69.34 ± 0.7996
4 250 66.04 ± 1.5487 72.50 ± 1.0886
5 300 78.44 ± 0.4442 84.02 ± 1.9981
11. Effect of Methanolic extract of Mimosa arenosa on Protein
Denaturation
S. No
Concentration
(µg/ml)
% Inhibition of Protein Denaturation (Mean ± SEM)
SD (Indomethacin) MME
1 100 40.26 ± 1.1224 43.12 ± 1.4263
2 150 44.64 ± 0.9033 49.76 ± 1.1219
3 200 50.75 ± 1.7243 55.25 ± 0.4200
4 250 62.51 ± 1.5542 69.40 ± 1.4322
5 300 72.47 ± 0.9447 76.92 ± 1.3537
12. Ex-Vivo Anti-Asthmatic effect of MME
S. No
Dose of
Histamine
(in µg/ml)
Log Dose of
Histamine
%
Contraction of
Histamine
% Contraction of
HIST + MME
% Contraction of
CPM
1 10 1 76.66 66.66 55.44
2 20 1.3 83.33 81.48 69.66
3 40 1.6 90 88.88 81.13
4 80 1.9 100 100 100
13. DISCUSSION AND CONCLUSION:
The % inhibition of heat induced haemolysis and protein denaturation of methanolic extract
of Mimosa were increased when compared to that of the standard group.
The dose response curve for histamine in the presence of methanolic extract of Mimosa
shifted towards right hand side which indicates the anti-asthmatic activity of Mimosa.
Thus from the present study, it was concluded that the Methanolic extract of Mimosa
arenosa whole plant shows significant anti-inflammatory and anti-asthmatic activities.
Further research work is needed to be done to evaluate its clear mechanism of action.
REFERENCES:
Mahesh S, Paschapur, Patil MB, Ravi Kumar and Sachin. “Evaluation of anti-inflammatory
activity of ethanolic extract of Borassus flabellifer L. male flowers in experimental animals”.
Journal of Medicinal Plant Research. 2009; 3: 49-54.
Suralkar M. M. and Sarda P. P. “Antihistaminic and Antispasmodic Potential of Pongamia
pinnata”. International Journal of Pharmaceutical and Phytopharmacological Research.
2014; 3(2): 34 – 38.
