EVALUATION OF ANTIOXIDANT AND ANTIULCER
ACTIVITY OF ROOTS OF ZIZYPHUS JUJUBA LINN
GUIDANCE
Mr. P V SREEDHAR REDDY,
SUBMITTED BY: [Link].,Ph.D.
Asso. Professor,
NAME : GUNDABATHINA ASWITHA Department of Pharmacology
HT NO. : 20DK1S0106
SAASTRA COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH
Varigonda(V),[Link],SPSR Nellore
INTRODUCTION
• Peptic ulcers are a form of acid peptic [Link] can be classified according to their location and
other factors.
• It is break in inner lining of stomach and duodenum.
• They are two types
• A) Gastric ulcer – It occurs in the stomach.
• B)Duodenal ulcer-It occurs in the first part
of small intestine.
• Common cause is Helicobacter pylori
• NSAID’S.
• Common symptoms for ulcer is stomach
• Pain ,burning sensation, vomiting.
Microscopic view of peptic ulcer
PATHOPHYSIOLOGY AND ETIOLOGY
ANTIOXIDANT AND ANTI-ULCER ACTIVITY
• Oxidation is a chemical reaction that can produce free radicals there by leading to chain reaction
that may damage cell of organism.
• Antioxidant are the compounds that inhibit oxidation.
• It is a substance that protect cells from the damage caused by the free radical scavangers.
• Antioxidants like ascorbic acid ,hydrogen peroxide,DPPH radical scavenger activity
• Anti ulcer activity is to treat ulcers especially ulcers of the wall of stomach and duodenum by
antiulcer drugs by inhibition of gastric acid production,neutralization of acid from acid injury.
• Drugs like proton pump inhibitors eg:omeprazole,pantoprazolelansoprazole;H2 receptor blockers
eg:cimetidine, ranitidine;Ulcer protectives eg: sucralfate;Antibiotics eg:metronidazole,amoxicillin.
PLANT PROFILE
• Botanical name : Myrica nagi
• Family : Myricaceae
• VERNACULAR NAMES
• Hindi : Kaiphal
• Kannada : Kirishivani
• Malayalam : Maruta
• Sanskrit : katphala
• Tamil : Chavviyaci
• Telugu : Kaidarayamu
• Marathi : Kayaphal
Phytochemistry:
The bark is yellow and contains the chemical substances myricetin, myricitrin
and glycosides. Myricetin is important chemical constituents found in Myrica
nagi. Myricetin 50, yellow-beige powder crystalline powder, a flavonol,
consisting of 3-hydroxyflavone backbone and 6 hydroxyl groups has been
extracted from the leaves and fruits of the species..
Traditional uses:
-Powdered bark used as dressing for old wounds and ulcers.
- In Cambodia, bark is used for dysentery and gingivitis.
- Some Benue tribes use the leaves as ingredient in a prescription for
gonorrhoea
- Pounded leaves are applied as dressing to wounds. Leaves, in plaster
form, used in strangury. Paste made from tender leaves and twigs applied
to boils, abscesses and carbuncles to promote suppuration.
- Dried and ripe fruit used is mildly laxative; also, used as expectorant.
- In China, fruit used for coughs. Also, used for insomnia and anxiety
AIM & OBJECTIVE
AIM :
The aim of the present study was designed to evaluation of antioxidant and antiulcer activity of leaves of
myrica nagi
OBJECTIVES :
Collection of Plant material (leaves of myrica nagi).
Preliminary phytochemical screening of crude extract.
Evaluation of antioxidant activity by DPPH,reducing power and hydrogen peroxide free radical
scavenging method.
Evaluation of antiulcer activity in ethanol induced ulcer model and aspirin-induced gastric lesions in rats.
Treatment protocol with extracts.
Selection, grouping and acclimatization of animals.
Parameters to study ulcer index and % protection.
MATERIALS AND METHODS
Plant Collection and Authentication:
• The root of Myrica nagi was collected from the tirumala forests, Tirupati,Chittoor district, A.P,
India in the month of August 2020. The plant sample was further verified and authentified by Dr.
K. Madhava Chetty, Department of Botany, Sri Venkateswara University, Tirupati.
Experimental animals:
• Healthy adult female albino rats of wistar strain were procured from raghavendra enterprises,
Bangalore weighing between 150-200 g were used in the present study.
Extraction:
• The leaves of Myrica nagi were shade dried and reduced to coarse powder in a mechanical
grinder . The powdered material obtained was then subjected to maceration using water for 72 hrs
and the aqueous root was used for further study.
Determination of preliminary phytochemical screenings:
1. Test for Carbohydrates : Molisch’s test
2. Test for reducing sugars : Benedict’s test,Fehling’s test
3. Test for Alkaloids :Dragendorff’s test,Mayer’s test,Hager’s test,Wagner’s test
4. Test for phytosterols :Salkowski’s test,Libermann Burchard’s test
5. Test for Phenols :Ferric chloride test
6. Test for Tannins :Gelatin test
7. Test for Flavonoids :Alkaline reagent test, Lead acetate test,Shinoda test
8. Test for Prpteins and Aminocids :Xanthoprotein test, Ninhydrine test,Biuret test
Experimental study:
1. Throughout the study, rats were housed in temperature-controlled
rooms with12-h light 12-h dark cycle and were free access to food
and water.
2. Animals were divided into four groups as control (n=6), Aspirin
20mg/[Link],p o (n=6), Aspirin20 mg/kg+OMZ 50mg/kg (n=6),
Aqu root extract 150mg/kg+Aspirin 20 mg/kg(n=6), Aqu root
extract 250mg/kg+Aspirin 20mg/kg (n=6) where n was the number
of animals included in the study.
3. Animals were divided into four groups as control (n=6),Ethanol
1ml/[Link] (n=6),Ethanol 1ml/[Link]+OMZ 50mg/kg
(n=6),Aqueous leaves extract 150mg/kg+ethanol [Link]
(n=6),Aqueous leaves extract 250mg/kg+ethanol 1ml/[Link](n=6)
where n was the number of animals included in the study.
TREATMENT PROTOCAL
• GROUP I : Negative control group
(n=6, the animals were given Aspirin 20mg/kg only)
• GROUP II : positive control group
(n=6, the animals were given OMZ 50mg/kg+Aspirin 20mg/kg for 2Hr).
• GROUP III : Treatment group
(n=6, the animals were given ALMN150 mg/kg+Aspirin20mg/kg for 2Hr).
• GROUP IV : Treatment group
(n=6, the animals were given ALMN 250mg/kg+Aspirin 20mg/kg for 2 Hr).
TREATMENT PROTOCOL
• GROUP I : Negative control group
(n=6, the animals were given Ethanol 1mg/kg only)
• GROUP II : positive control group
(n=6, the animals were given OMZ 50mg/kg+Ethanol 1ml/kg for 2Hr).
• GROUP III : Treatment group
(n=6, the animals were given AEZJ150 mg/kg+Ehanol 1ml/kg for
2Hr).
• GROUP IV : Treatment group
(n=6, the animals were given AEZJ 250mgkg+Ethanol 1ml/kg for 2 Hr).
EXPERIMENTAL DESIGN:
Rats were treated as per the treatment protocol. The protocol was approved by the Institute’s Animal
Ethical Committee. Body weights of these rats were monitored sequentially in control and
experimental animals for a period of 28 days. Theywere sacrificed on day 29. the stomach were
opened along the greater curvature,washed gently with saline water to remove gastrin content and
blood clots and examined 10X magnifier lence to assess the formation of ulcer.
Parameters to study:
[Link] index : The ulcer index was calculated by adding the total number of ulcers per stomachand
total severity of ulcer per stomach
formulae : UI =UN +U S +UP * 10-1
[Link] inhibition: The percentage inhibition was calculated as
% inhibition = ulcer index control – ulcer indextest* 100
Ulcer indexcontroi
[Link] analysis: Results are expressed as mean ± SEM and were performed one way analysis of ANOVA method.
RESULTS
PHYTOCHEMICAL ANALYSIS OF ALMN
[Link] PHYTOCHEMICAL TEST AQUEOUS ROOT EXTRACT
OF ZIZIPHUS JUJUBA LINN
1 Test for alkaloids +ve
2 Test for anthraquinones -ve
3 Test for carbohydrates +ve
4 Test for flavonoids +ve
5 Test for glycosides +ve
6 Test for proteins +ve
7 Test for tannins +ve
8 Test for terpinoids +ve
10 Test for coumarines +ve
COMPARISION OF PERCENTAGE INHIBITION OF AQUEOUS ROOT EXTRACT OF
MYRICA NAGI WITH THE STANDARD ASCORBIC ACID
DPPH
100
A
b 90
s
o 80
r
b 70
a
n 60
c Standard Ascorbic acid
e 50
[Link]
a 40
t
30
5
1 20
7
10
n
m 0
100 120 140 160 180 200
Concentration µg/ml
The ALMN showed potent antioxidant activity with the percentage inhibition ranging from
30% to 90 % at various concentrations and is compared with the standard ascorbic acid
COMPARISION OF PERCENTAGE INHIBITION OF AQUEOUS ROOT EXTRACT OF MYRICA
NAGI WITH THE STANDARD ASCORBIC ACID BY H 2 O 2 METHOD
H2O2
A
b 120
s
o
r 100
b
a
n 80
c Standard Ascorbic Acid
e Aq. Extract
2 60
3
0 40
n
m
20
0
0.1 0.2 0.4 0.600000000000001 0.8
Concentration µg/ml
The ALMN showed maximum percentage inhibition at 0.8 mcg/ml i.e 87% and is
comparable with thestandard which indicates that the extract is almost equipotent in
scavenging the free radicals as standard ascorbic acid
Anti-ulcer activity of ALMN. At the dose of 150 mg/kg bwtand 250
mg/[Link] in Ethanol induced ulcer model
GROUPS TREATMENT MEAN ULCER INDEX % INHIBIT ION OF
± SEM ULCER
INDEX
No inhibition
GROUP 1 NEGATIVE CONTROL 2.5* ± 0.341
( ETHANOL 1ML/KG BWT)
GROUP 2 POSITIVE CONTROL 0.5* ± 0.129
(OMEPRAZOLE 50 MG/KG BWT) 80
GROUP 3 AQUEOUS ROOT EXTRACT ( 150 1.583**±0.374
MG/KG BWT 36.68
GROUP 4 AQUEOUS ROOT EXTRACT ( 250 0.75*** ± 0.111
MG/KG BWT 70.00
Treatment schedule – Anti-ulcer activity of ARZJ in
Aspirin induced ulcer model.
S .no Groups Treatment Mean ulcer % Inhibit ion
index ±SEM of index
1 Group 1 Negative control 2.167*U±0.307 No inhibition
(Aspirin 20 mg/kg
bwt )
2 Group 2 Positive control 0.416*±0.153 80.77
(Omeprazol
50mg/kg)
3 Group 3 Aqueous root extract 0.583*±0.153 73.08
(150 mg/kg)
4 Group 4 Aqueous root extract 0.5*±0.129 76.92
(250 mg/kg)
A) NEGATIVE CONTROL B) POSITIVE-CONTROL
( OMEPRAZOLE 50 MG/KG)
(ASPIRIN
20MG/[Link])
C)AQUEOUS ROOT D)AQUEOUS ROOT EXTRACT
EXTRACT
(150 MG/[Link]) ( 250 MG/KG BWT)
Treatment schedule-Anti-ulcer activity of ALMN in
Ethanol induced ulcer model
[Link] GROUPS TREATMENT MEAN ULCER INDEX % INHIBITION OF
±SEM ULCER INDEX
1 Group1 Negative control 2.5*±0.341 No inhibition
(ethanol 1ml/[Link])
2 Group2 Positive control 0.5*±0.129 80
(omeprazole
50mg/[Link])
3 Group3 Aqueous root extract 1.583*±0.374 36.68
(150mg/[Link])
4 Group4 Aqueous root extract 0.75*±0.111 70.00
(250 mg/[Link])
a) Negative control b) Positive control
(Ethanol 20 mg/kg bwt) (Omeprazole 50 mg/kg bwt)
c) Aqueous root extract
d) Aqueous root extract
(150 mg/kg bwt)
(250 mg/kg bwt)
DISCUSSION
• Gastric and duodenal ulcers are commom problem today because of defencive
and offencive factors.
• A wide variety of pharmacodynamics effect such as anti
inflammatory,antimicrobial,antiallergic activities attributed to flavonoids.
Flavonoids kown to inhibit several enzymes like alkaline phosphate,
lipases,H˖ATPase and Na˖/k˖ ATPse.
• The ALMN showed inhibition of free [Link] ALMN at the dose of
150mg/kg bwt inhibited ulcer index by 36.68% and at the dose of 250mg/kg
bwt inhibited ulcer index by 70.00% .
• The ALMN at the dose of 250mg/kg bwt was found to impart better
antiulcerogenic activity when compared to the dose of 150mg/kg bwt.
• In ALMN presence of flavonoids in the extract suppress the aggressive factors
which may lead to the reduction of gastric ulceration.
• From this ALMN developed as therapeutic antioxidant and antiulcerogenic
agent.
CONCLUSION
• The results obtained from the qualitative phytochemical study reveals the presence of
alkaloids,carbohydrates, Phenolic compound,glycosides,sapponnins,tannins, and flavonoids.
• ALMN is a good antiulcer agent.
• It showed a strong antioxidant activity by inhibiting DPPH,H₂O₂,and reducing power activities
when compared with standard L-Ascorbic acid.
• ALMN shows a substantial and significant protection against gastric.
• Further studied may be required to isolate and characteristic various phyto constituent and the
study of the agent in long term use.
• The antiulcer activity long with its safety profile could make the root of ziziphus jujuba a good
candidate for the treatment of PUD in humans.
REFERENCES
• Subhose V,Srinivas P,Narayana [Link] principles of pharmaceutical science
in Ayurveda. Bull Indian InstHist Med Hyderabad.2005;35(2):83-89.
• Mukherjee PK,Shau M, Suresh [Link] Herbal [Link]
Pharmacist;1998(8):21-24.
• Tandon V,Kappor B,Gupta [Link] drug research in India,A trend analysis
using IJP as marker(1995-August2003).Indian Journal pharmacol
2004;36(2):99-100.
• Allison MC ,Howatson AG,Torrance CJ, Lee FD, Russell
[Link] associated with the use of nonsteroidal anti-
inflammatory drugs. N EngIJ Med1992;327:749-54.
• Lenz HJ,Ferrari-Taylor J,Isenberg [Link] and five percent ethanol are potent
stimulants of gastric acid secretion in [Link].1983;85:1082-
7.
• Naftali [Link] H,.myrica nagi Extract for the Treatment of chronic
Idiopathic Constipation’
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