Investigations on In-Vitro Anti-Inflammatory Activity Of

“INVESTIGATIONS ON IN-VITRO ANTI-INFLAMMATORY ACTIVITY OF

MIMOSA PUDICA LINN”

SYNOPSIS FOR

M.PHARM DISSERTATION

SUBMITTED TO

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA

BY

SUMAIYA SARVATH I M PHARM

DEPARTMENT OF PHARMACOGNOSY

AL-AMEEN COLLEGE OF PHARMACY

BANGALORE - 560027

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE, KARNATAKA.

ANNEXURE – II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / NAME OF THE CANDIDATE AND ADDRESS / SUMAIYA SARVATH
D/O M. ATHA ULLA KHAN
DOOR NO 4317, MASJID MOHALLA,SHETHALLY BEDI, BEHIND RHAMANIA GIRL’S HIGH SCHOOL.RAMANAGARAM-571511
BANGALORE
KARNATAKA.
2. / NAME OF THE INSTITUTION / AL- AMEEN COLLEGE OF PHARMACY
HOSUR ROAD, BANGALORE – 560027
3. / COURSE OF STUDY AND SUBJECT / M. PHARM - PHARMACOGNOSY
4. / DATE OF ADMISSION / JUNE – 2009
5. / TITLE OF THE TOPIC :
“INVESTIGATIONS ON IN-VITRO ANTI-INFLAMMATORY ACTIVITY OF MIMOSA PUDICA LINN”
6.0 / BRIEF RESUME OF THE INTENDED WORK :
6.1 / NEED OF STUDY
Mimosa pudica Linn (Mimosaceae) is a diffuse under shrub, commonly known as the “Touch-me-not” plant. The local name of M. pudica is lajvanti or chuimui. This plant is native of tropical America and Brazil but is cosmopolitan. M. pudica is an important herb in the Ayurvedic, unani, and Herbal medicine. In Ayurvedic and unani medicine, roots and leaves of M. pudica are commonly used as bitter, cooling, astringent, to treat inflammation, piles, leprosy, small pox, fistula, burning sensation. In ethnomedicine it used for ulcers, fever, headache, migraine, dysentery, diarrhea, to stop menstruation, conjunctivitis, cuts, wounds, odema, rheumatism, myalgia, skin diseases and kidney diseases. The roots are used as temporary birth control in India.1,2,3
The extracts of roots and leaves of M pudica have been reported for its pharmacological activities such as, ovulation reduction,4 anticonvulsant,5 antidepressant,6 antidiabetic,7 antimicrobial,8 wound healing,9 snake venom induced hyaluronidase and protease inhibition, snake venom neutralization,10 and antioxidant activities11.
The phytochemical constituents of M pudica: are alkaloids, steroids, tannins, triterpenes, flavonoids, glycosides, quinines, phenols, saponin, coumarin, c-glycosylflavones5,8.
M. pudica has been widely used to treat oedema, rheumatism, myalgia and inflammation in traditional and tribal medicine, however no scientific investigations are reported for its anti-inflammatory properties.
The inflammatory response involves a complex array of enzyme activation, mediator release, fluid extravasations, cell migration, tissue breakdown and repair which are aimed at host defense and usually activated in most disease condition12. Several phenomena alter the antigenicity of endogenous proteins, including protein denaturation and glycosylation. Protein denaturation may occur during chronic inflammatory phenomena in vivo and albumin denaturation was observed in patients with rheumatic diseases and in rats with inflammatory lesions13. Lysosomal enzymes released during inflammation have been implied in acute or chronic inflammation. Many of the NSAIDs such as diclofenac act by inhibiting these lysosomal enzymes or by stabilizing the lysosomal membrane. Since the membrane of RBC is structurally similar to lysosomal membrane, the effect of any substance on stabilization of RBC membrane may be extrapolated to the stabilization of lysosomal membrane14,15.
It has been reported that leucocyte proteinases play an important role in the development of tissue damage during inflammatory reactions and significant level of protection was provided by proteinase inhibitors. Hence inhibition of albumin denaturation, RBC membrane stabilization and protease inhibition afford. Protection against chronic inflammatory conditions15.
We propose to study anti inflammatory activity of M. pudica by studying the effects of the different extracts on protein denaturation inhibition, proteinase inhibition and human RBCmembrane stabilisation, in-vitro.
Reactive oxygen species (ROS)play an important role in the pathogenesis of inflammatory diseases. Antioxidants capable of scavenging ROS are expected to improve these disorders16. Hence we propose to study the antioxidant properties of the extracts for its ferric reducimg anti-oxidant power and DPPH radical scavenging activity.
6.2 /

LITERATURE REVIEW

Mimosa pudica Linn (Mimosaceae) is a diffuse undershrub of cosmopoliton distribution. It is used to treat various ailments such as piles,fistula, glandular swellings, inflammation, asthama, leucorrhoea, leprosy, jaundice ete. in traditional and ethnomedicine1,2,3.
The roots and leaves of M. pudica have been reported for its pharmacological activities such as, aphrodisiac,3 ovulation reduction,4 anticonvulsant,5 antidepressant,6 antidiabetic,7 antimicrobial,8 wound healing,9 snake venom induced hyaluronidase and protease inhibition, snake venom neutralization,10 and antioxidant activities11.
Orally administered ethanolic extract of roots of M. pudica significantly increased libido and testosterone levels in male mice3. Root powder administered orally caused significant reduction in number of normal ova and a significant increase in number of degenerated ova in female albino rats4. An aqueous decoction of leaves of the plant given i.p protected mice against pentylentetrazol and strychnine-induced seizures5. Aqueous extract of M. pudica reduced immobility in the forced swimming test and increased rate of reinforcers in the differential reinforcement of low rates of response at 72 sec Suggesting its antidepressant effects in rats6. Ethanolic extracts of leaves of M. pudica showed a significant decrease in blood glucose level in alloxan-induced diabetes in albino rats7.
Methanolic extracts of leaves of M. pudica excihibited antimicrobial activity against Aspergillus fumigatus, Citrobacter divergens and Klebsiella pneumonia by well diffusion method. This extract showed presence of terpenoids, flavonoids, steroids, glycosides, alkaloids, quinines, phenols, tannins, saponins and coumarins8.
Methanolic extracts of root and shoot of M. pudica Linn showed vary good wound healing activity in Wistar albino rats as compared to the standard drug gentamicin9. The aqueous root extract of M. pudica inhibited the hyaluronidase and protease activities in Indian snake venomes10. Dichloromethane (DCM) and methanol extracts of stems of M. pudica exhibited considerable anti-oxidant and bacteriostatic activity against Bacillus subtilis, E.coli, Staphylococcus aureus, and Pseudomonas auruginosa. However, no significant toxicity was observed towards brine shrimps11.
Inflammatory response involves a complex series of reactions human red blood cells (HRBC) membrane stabilization in vitro as an indicator of anti-inflammatory activity were studied for n-hexane, chloroform, ethyl acetate and methanolic extract of leaves of S.cochinchnensis. The methanolic extract show 67% of HRBC in hypotonic solution as against 74% protection showed by diclophenac12.
Protein changes such as albumin denaturation and glycosylation patterns are altered during chronic inflammatory conditions. Substance capable of preventing these changes could be of potential value in conditions like rheumatism. Saso et.al. have studied the effect of several exogeneous and endogeneous compounds such as fish oil, bile salt and fatty acids on inhibition of protein denaturation inhibition13.
The powder drug of leaves of Abutilon indicum were extracted successively with several solvents; ethanolic, chloroform and aqueous extract were screened for in vitro anti-inflammatory activity. Both extracts showed a biphasic effect in prevention of hypotonicity induced HRBC membrane lysis indicative of anti-inflammatory activity14.
A polyherbal preparation has been evaluated for antiarthritic and anti-inflammatory activity in adjuvant induced non-established and established arthtitis in-vivo. The preparation also has been tested for protection against protein denaturation, RBC membrane and proteinase activity in-vitro. The preparation showed anti-inflammatory activity in all in-vivo and in-vitro models tested15.
Reactive oxygen species (ROS) play an important role in the pathogenesis of inflammatory diseases. Antioxidants which can scavange Reactive Oxygen Species are expected to improve such disorders. The anti-inflammatory and anti-oxidant activities of the aqueous and methonolic extracts of Cassia fistula Linn bark were evaluated in acute and chronic models in Wistar albino rats. Both Extracts showed significant anti-oxidant effects. The extracts significantly inhibited CCL4 and FeSO 4 initiated lipid-peroxidation in rat liver and kidney homogenates as well as scavenged DPPH, Nitric oxide and hydroxy radicals in in-vitro assays16 .
6.3 / AIMS AND OBJECTIVES OF THE STUDY
AIM:-
To Investigate the phytochemical composition and In-vitro anti-inflammatory activity of Mimosa pudica Linn.
/ OBJECTIVES: -
1.  Collection and Pharmacognostic evaluation of the drug.
2.  Preparation of various extracts by successive solvent extraction.
3.  Standardization of In-vitro Bioassays
a.  Standardization of membrane stabilization activity using human RBC.
b.  Standardization of protein denaturation inhibition activity.
c.  Standardization of protease inhibitory activity.
4.  Determination of these activites for the extracts prepared.
5.  The antioxidant activity of the different extracts.
6.  Phytochemical investigation of different extracts.
7.0
7.1 / MATERIALS AND METHODS
SOURCES OF DATA
§  Al-Ameen college of Pharmacy Information Center, Bangalore.
§  Literature survey.
§  Journal and Publications.
§  Web search on - Science direct, Pub med, Medline, interscience, elsevier, highwire press, Ethnobiomed, Google.
§  J-Gate@HELINET search.
PLACE OF WORK:-
·  Al-Ameen college of Pharmacy, Bangalore.
7.2 /

METHOD OF COLLECTION OF DATA

1.  The plant will be collected and authenticated at IIHR, Bangalore.
2.  Pharmacognostic Evaluation -Macroscopy, Microscopy and Proximate value determination.
3.  Successive solvent extraction of roots, aerial parts and entire plant using Petroleum ether, Chloroform and Alcohol.
4.  In-vitro Bioassays for membrane stabilization activity12, protein denaturation inhibition13 and protease inhibition15.
5.  Antioxidant activity of extracts showing good anti-inflammatory activity from the In-vitro assay.
6.  Phytochemical investigations of the extracts showimg activity.
7.3 / Does the study require any investigations or interventions to be conducted on patient or other humans or animals? If so please describe briefly.
-NA-
7.4 / Has ethical clearance been obtained from your institution in case of the above query?
-NA-
8.0 / REFERENCES:
1.  Kritikar and Basu. Indian Medicinal Plants, International Book Distributiors,
Derhadun, India 1987;2:915-917.
2.  Cherly L. Ethnomedicine used in Trinidad and Tobago for reproductive problems. J.Ethnobiology and Ethnomedicine 2007; 3:1-12.
3.  Milind P, Anupam P. Aprodisiac Activity of Roots of Mimosa pudica Linn. Ethanolic Extract in Mice, Inter.J .Pharmaceuti.Sci. Nanotechnol 2009; 2(1):477-486.
4.  Valsala S, Karpagaganapathy PR. Effect of Mimosa pudica root powder on oestrous cycle and ovulation in cycling female albino rat, Rattus norvegicus. Phytother Res 2002;16(2):190-192.
5.  Bum EN, Dawack DL, Schmutz M, Rakotonirina A, Rakotonirina SV, Portet C, Jeker A, Olpe HR, Herrling P. Anticonvulsant activity of Mimosa pudica decoction. Fitoterapia 2004;75:309-314.
6.  Molina M, Contreas CM, Tellez AP. Mimosa pudica may posses antidepressant action in the rat. Phytomedicine 1999;6(5):319-323.
7.  Sutar NG, Sutar UN, Behera BC. Antidiabetic Activity of the Leaves of Mimosa pudica Linn in albino rats. J.Herbal Medicine and Toxicology 2009;3(1):123-126.
8.  Gandiraja N, Sriram S, Meenaa V, Srilakshmi JK, Sasikumar C, Rajeswari R. Phytochemical Screening and Antimicrobial Activity of the Plant Extracts of Mimosa pudica L.Against selected Microbes. Ethnobotanical Leaflets 2009;13:618-24.
9.  Kannan S, Jesuraj SAV, Jeeva KES, Saminathan K, Suthakaran R, Ravi KM, Parimala DB. Wound healing activity of Mimosa pudica Linn formulation. Inter. J. Pharmtech Research 2009; 1:1554-1558.
10.  Girish KS, Mohanakumari HP, Nagaraju S, Vishwanath BS, Kemparaju K. Hyaluronidase and Protease activities from Indian snake venoms: neutralization by Mimosa pudica root extract. Fitoterapia 2004;75:378-380.
11.  Genest S, Kerr C, Shah A, Rahman MM, Saif GMM, Nigam P, Nahar L, Sarker SD. Comparative bioactivity studies on two Mimosa species. Boletin Latinoamericano y del caribe de plantas Medicinales y Aromaticas 2008;7(1):38-43.
12.  Rajendran V, Lakshmi KS. In vitro and in vivo anti-inflammatory activity of leaves of symplocos cochinchnesis(lour) Moore ssp laurina. J.Pharmacol 2008;3:121-124.
13.  Luciano S. Giovanni V, Maria LC, Eleonora M, Laura B, Gabriela M, Claudio P, Eleonora G, Bruno S. Inhibition of Protien Denaturation by Fatty Acids,Bile salts and other Natural Substances: A New Hypothesis for the Mechanism of Action of Fish Oil in Rheumatic Diseases. Jpn. J. Pharmacol 1999;79:89-99.
14.  Rajurkar R, Jain R, Matake N, Aswar P, Khadbadi SS. Ati-inflammatory Action of Abutilon indicum(L) Sweet Leaves by HRBC Membrane Stabilization. Res J. Pharm. Tech. 2009;2(2):415-416.
15.  Chatterjee S, Das SN. Anti-arthritic and Anti-inflammatory effect of a poly-herbal drug (EASE@) : its mechanisam of action. Indian.J.Pharmacology 1996;28:166-119.
16.  Ilavarassan R, Mallika M, Venkataraman S. Anti-inflammatory and Antioxidant Activities of Cassia Fistula Linn bark extracts. Afr. J. Trad. CAM 2005;2(1):70-85.
9. / SIGNATURE OF THE CANDIDATE:
10. / REMARKS OF THE GUIDE: / RECOMMENDED FOR RESEARCH AND SUBMISSION
11. / NAME AND DESIGNATION OF THE GUIDE: / Mrs. BHUVANESHWARI J
Lecturer, Department of Pharmacognosy,
Al-Ameen College of Pharmacy,
Bangalore.
11.1. / SIGNATURE:
11.2 / HEAD OF THE DEPARTMENT: / Dr. SALMA KHANAM
PROF. AND H.O.D.
Department of Pharmacognosy,
Al-Ameen College of Pharmacy,
Bangalore.
11.3 / SIGNATURE:
11.4 / PRINCIPAL: / Prof. B.G. SHIVANANDA
Al-Ameen College of Pharmacy,
Hosur road, Bangalore.
11.5 / REMARKS OF THE PRINCIPAL: / RECOMMENDED FOR RESEARCH AND SUBMISSION OF DISSERTATION
11.6 / SIGNATURE:

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