Antioxidant, Anti-Inflammatory and Anti-Nociceptive Effects of Ammannia

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Journal of Ethnopharmacology 140 (2012) 230–233

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Journal of Ethnopharmacology

jo urnal homepage: www.elsevier.com/locate/jethpharm

Antioxidant, anti-inﬂammatory and anti-nociceptive effects of

Ammannia baccifera L. (Lythracceae), a folklore medicinal plant

a b a,∗

Nataraj Loganayaki , Perumal Siddhuraju , Sellamuthu Manian

Department of Botany, School of Life Sciences, Bharathiar University, Coimbatore 641046, TN, India

Department of Environmental Sciences, School of Life Sciences, Bharathiar University, Coimbatore 641046, TN, India

a r t i c l e i n f o

a b s t r a c t

Article history: Ethnopharmacological relevance: Ammannia baccifera L. has been reported as folklore remedy for the

Received 23 June 2011

treatment of inﬂammation and tumor in the state of Rajasthan, India.

Received in revised form 13 October 2011

Aim of the study: The present study was designed to investigate the antioxidant, anti-inﬂammatory and

Accepted 2 January 2012

anti-nociceptive effects of the methanol extract from the aerial parts of Ammannia baccifera under in vitro

Available online 20 January 2012

and in vivo models.

•

Materials and methods: The in vitro antioxidant activity of the extract was measured using DPPH , super-

Keywords:

oxide, hydroxyl and nitric oxide radicals. The anti-inﬂammatory activity was evaluated using carrageenan

Ammannia baccifera

Anti-inﬂammatory induced paw edema. Analgesic activity of the methanol extract was estimated against acetic acid-induced

Antioxidant writhing and hot plate methods.

Paw edema Results: The IC50 value for free radical scavenging activity of this extract was signiﬁcantly superior over

Hot plate the positive standards butylated hydroxyl anisole (BHA) and rutin. The extract exhibited signiﬁcant anti-

inﬂammatory and analgesic activities at the dose of 100 and 200 mg/kg p.o. The analgesic effect of the

higher dose of the extract (200 mg/kg) was comparable with the standard drugs aspirin and morphine.

Conclusion: The present study scientiﬁcally validated the traditional use of this plant against inﬂamma- tion.

1. Introduction rhamnetin and its 3-rhamnosylglucoside were found to be present

in Ammannia multiﬂora (Balraj and Nagarajan, 1981). In the present

Ammannia baccifera L. belonging to the family Lythraceae is study, we investigated the analgesic and anti-inﬂammatory prop-

distributed commonly through out India, often as a weed in rice erties of the methanol extract of Ammannia baccifera using several

ﬁelds. In Ayurveda, the leaves are considered to have laxadive, experimental models with mice and rats for the purpose of validat-

stomachic, hepatopathy and aphrodisiac properties and are use- ing its ethnomedical use. Natural active principles, endowed with

ful for treating strangury. In Unani they are used as an appetizer anti-inﬂammatory activity, frequently exhibit antioxidant power;

and for the treatment of rheumatic pains and fevers. In Siddha the moreover many plant extracts interacting with several biosynthetic

whole plant is used in the treatment of glandular swellings, leuko- pathways of inﬂammation mediators in general possess antioxi-

rrhea, snake-bite poisoning abscesses, ulcers and polyuria. Leaves dant property (Speroni et al., 2006). For this reason, the extracts

contain lawsone as the major chemical constituent. The herb is were also assessed for antioxidant capacity, using different in vitro

reported to possess anti-typhoid, anti-tubercular and anti-tumor chemical methods.

properties (Parrotta, 2001; Khare, 2007). The different extracts of

the plant are reported to have antimicrobial activity against human

and plant pathogenic fungi (Ray et al., 2004). Four ﬂavonol and three 2. Materials and methods

ﬂavone glycosides quercetin 3-␤-d-glucoside, rutin, luteolin 7-␤-

d ␤ d

-glucoside, isorhamnetin 3-rutinoside, apigenin 7- - -glucoside, 2.1. Plant material

vitexin and kaempferol 3-rhamnoglucoside were isolated and char-

acterized from Ammannia coccinea (Graham et al., 1980). Free Aerial part of Ammannia baccifera were collected from local

areas of Coimbatore district, Tamil Nadu, India, authenticated

and deposited in the Botany Herbarium, Bharathiar University

∗ (BUH006148). The plant material was shade dried, powdered and

Corresponding author. Tel.: +91 9442170766; fax: +91 422 2422387.

E-mail address: manian [email protected] (S. Manian). used for methanol extraction.

N. Loganayaki et al. / Journal of Ethnopharmacology 140 (2012) 230–233 231

2.2. Preparation of extract Group 1: received the vehicle saline (10 ml/kg p.o.).

Group 2: treated with standard drug.

The plant material was successively extracted with petroleum Group 3: treated with methanol extract of Ammannia baccifera

ether (dewaxing) and methanol using soxhlet apparatus. The sol- (100 mg/kg p.o).

vent was removed and used for further experiment. Group 4: treated with methanol extract of Ammannia baccifera

(200 mg/kg p.o).

2.3. Estimation of total phenolics, tannins and ﬂavonoids

2.5.4.1. Acetic acid-induced writhing. The peripheral analgesic

drug, aspirin was used as positive control (25 mg/kg, p.o.). Acetic

The total phenolic content and tannins of the extract was deter-

acid-induced writhing was determined using the method of

mined and calculated as g gallic acid equivalents (GAE) from the

Dongmo et al. (2005).

calibration curve (Makkar, 1999). The total ﬂavonoid content of

sample extract was determined following a colorimetric method

2.5.4.2. Hot-plate test in mice. The central analgesic drug morphine

and values were expressed as g rutin equivalent (RE)/100 g of

(5 mg/kg p.o.), was used as positive control. Latency to exhibit

extract (Zhishen et al., 1999).

antinociceptive responses was determined by MacDonald et al.

(1946). Latency was recorded 15, 30, 45, 60, 90 and 120 min after

2.4. In vitro antioxidant activity administration of the test substances.

•

The free radical scavenging assays include DPPH (Blois, 1958), 2.6. Statistical analysis

Superoxide (Beauchamp and Fridovich, 1971), Hydroxyl (Klein

et al., 1991) and Nitric oxide (Sreejayan and Rao, 1997) scavenging The results were expressed as mean ± S. E. M. Statistical anal-

activities were measured using standard procedures. ysis was carried out by analysis of variance (ANOVA) followed by

Dunnett’s test. All calculations were performed using: SPSS, version

11.0.

2.5. In vivo studies

3. Results

2.5.1. Animals

Male Swiss albino mice (25–30 g) and Wistar albino rats

The quantitative estimations of total phenolics, tannins and

(150–175 g) were used for the study. All animal experiments were

ﬂavonoids from methanol extract of Ammannia baccifera revealed

conducted with the permission from Institutional Ethical Commit-

that the extract has 95.7 ± 1.6, 6.2 ± 4.2 and 43.3 ± 0.1 g/100 g

tee (IAEC No. KMCRET/Ph.D/01/2010-11).

extract of total phenolics, tannins and ﬂavonoids, respectively.

The antioxidant activity of methanol extract of Ammannia bac-

•

2.5.2. Acute toxicity

cifera was evaluated using DPPH , super oxide, hydroxyl and

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Acute oral toxicity studies were performed according to OECD

nitric oxide radicals. The IC50 value of DPPH scavenging activ-

421 (Organization for Economic Co-operation and Development).

ity of methanol extract of Ammannia baccifera was 8.3 ␮g/mL.

This value was lower than those of positive standards BHA

(9.7 ␮g/mL) and rutin (17.4 ␮g/mL). Like that, super oxide and

2.5.3. Carrageenan induced paw edema

hydroxyl radical scavenging activities also methanol extract of

For the experiment, the male Wistar rats (150–175 g) were

Ammannia baccifera (25.9 ± 2.1 and 54.6 ± 1.3 ␮g/mL, respectively)

divided into four groups (n = 6). The ﬁrst group received distilled

exhibited higher radical scavenging activity over positive stan-

water (10 ml/kg p.o.), while the second group was treated with

± ± ␮

dards BHA (79.6 1.3 and 75.7 3.7 g/mL, respectively) and rutin

standard drug indomethacin (10 mg/kg p.o.). The third and the

(62.3 ± 4.7 and 66.3 ± 2.9 ␮g/mL, respectively). However in nitric

fourth groups were administered with the methanol extract of

oxide scavenging activity, methanol extract of Ammannia bac-

Ammannia baccifera (100 and 200 mg/kg p.o. respectively). Acute

± ␮

cifera showed higher radical scavenging activity (98.6 5.7 g/mL)

inﬂammation was induced and paw volume was measured imme-

over BHA (113.9 ± 8.1 ␮g/mL) and comparable activity with rutin

diately at 1 h, 2 h, 3 h, 4 h, and 5 h after the carrageenan injection,

(94.7 ± 6.3 ␮g/mL).

by using digital Lethylsmometer (Winter and Poster, 1957).

The extract did not alter the general behavior and failed to pro-

duce any mortality even at the highest dose (2000 mg/kg, p.o.)

2.5.4. Analgesic activity

studied after 3 days and found to be safe.

For the experiment, Swiss albino mice (20–25 g) were divided

The inhibitory effects of Ammannia baccifera and endomethacin

into four groups (n = 6).

on carrageenon induced paw edema are reported in Table 1. For

Table 1

Anti-inﬂammatory effects of methanol extract from Ammannia baccifera on carrageenan induced paw edema in rats.

Dose Edema rate

Treatment (mg/kg b.w.) 1 h 2 h 3 h 4 h 5 h

Vehicle 26.8 ± 7.7 35.4 ± 10.2 49.9 ± 9.2 66.1 ± 13.6 81.7 ± 10.9

* ** * *

Indomethacin 25 23.2 ± 6.4 27.2 ± 9.6 27.7 ± 7.4 29.7 ± 9.9 30.3 ± 12.8

(13.4%) (23.2%) (44.4%) (55.1%) (62.9%)

Extract 100 24.4 ± 5.1 29.9 ± 4.7 37.3 ± 16.7 44.9 ± 19.2 44.8 ± 15.8

(8.9%) (15.5%) (25.2%) (32.0%) (43.2%)

* ** * *

200 23.9 ± 3.4 26.6 ± 5.7 28.9 ± 8.5 30.1 ± 7.4 33.2 ± 9.9

(10.8%) (24.8%) (42.0%) (54.4%) (59.3%)

The data represent the mean SEM (n = 6). Given in the brackets are percent decrease of edema over control values. Control (vehicle) – distilled water.

p < 0.05 compared to corresponding control.

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