Preliminary screening of methanolic extracts of Celastrus paniculatus and Tecomella undulata for analgesic and anti-inflammatory activities

ELSEVIER SCIENCE IRELAND

Journal

of Ethnopharmacology

42

( 1994)

193-198

Preliminary screening of methanolic extracts of Celastrus paniculatus and Tecomella undulata for analgesic and antiinflammatory activities Fayyaz Ahmad, Rafeeq Alam Khan*, Shahid Rasheed Departmentof Pharmacology,Faculty of Pharmacy, University (Received

26 April

1993; revision received 5 January

of Karachi,

Kurachi-75270,

1994: accepted 2.5 March

Pakisran

1994)

Abstract

Flowers of Celastruspaniculatus and whole plant of Tecomella undulata were extracted individually in absolute methanol. Using the hot water tail immersion test in mice and carrageenan induced pedal edema in rats. both extracts were tested for their oral analgesic and anti-inflammatory potentials. Results showed that C. paniculatus had both analgesic and anti-inflammatory activities, while T. undulata had only analgesic potential when compared with aspirin. Key

words:

Celastruspaniculatus; Tecomella undukata; Methanolic extract; Analgesic testing; Anti-inflammatory

1. Introduction Celastrus paniculatus

Willd. (Celastraceae) and (Roxb.) Seeman (Bignoniaceae) are widely distributed medicinal plants in various regions of southeast Asia (Riaz, 1989). Both have been known as master medicinal plants from ancient days in Ayurvedic medicinal practice but experimental work on these plants was not instituted before 1927 (Nadkarni, 1927). Drugs currently in use for the management of pain and inflammatory conditions are either narcotics (e.g. opioids). non-narcotics (e.g. salycylates) or glucocorticoid-like (e.g. hydrocortisone). All these drugs present well-known side-effects and toxic effects on chronic administration. It is Tecomelfa

undulata

* Corresponding 0378-8741/94/$07.00 SSDI

author.

0

0378-8741(94)01126-K

1994 Elsevier Science Ireland

testing

therefore essential that efforts should be made to study promising medicinal plants so as to develop new or more safe drugs (Ikram, 1983). Plants still present a large untrapped source of structurally novel compounds that might serve as a lead for the development of novel drugs (Ibn al-Baytar and Din Abdullah Bin, 1197- 1248). The lack of safe and effective analgesic and antiinflammatory drugs now in use prompted the present study, in which C. paniculatus and T. undulata were selected for their reported biological activities in ancient Ayurvedic books. The oil extracted from the seeds of C. paniculatus has been shown to exert tranquilizing and sedative effects on rats, cats and dogs (Seth et al., 1963). A partially purified aqueous extract of T. undulutu produced a moderate relaxation of the tone of dog intestine with no significant effect on

Ltd. All rights reserved

F. Ahmad er al. /J.

194

blood pressure or respiration (Bhattacharya 1971).

et al.,

2. Materials and methods 2.1. Plant material and drugs Plants were collected from areas around Karachi during July and August 1991. Both taxa were identified with the help of Flora of Pakistan (Nasir and Ali, 1977). Voucher specimens Ahmad (1080KUH and 4945KUH) were deposited in the Karachi University Herbarium, Department of Botany, University of Karachi. Flowers of C. paniculatus and whole plants of T. undulata were dried in shade at 21-30°C for 15-30 days. These parts were then chopped and ground. Approximately 1 kg of ground plant material was soaked in 500 ml absolute methanol for 6 weeks. The alcoholic extracts were then evaporated under reduced pressure using a rotary evaporator (Eyela) and then freeze-dried and stored at -30°C for further evaluation. In terms of dry starting material, the yield of the methanol extract of C. paniculatus flowers was 5.9X, while the yield of the extract of whole plant T. undulata was 6.2%. Acetylsalicylic acid 300 mg, and the extracts of C. paniculatus and T. undulata in the quantities of 300,500 and 1000 mg were homogenized in appropriate amounts of 1.5% aqueous gum tragacanth. The suspensions were then administered orally (mice, 10 ml/kg; rats, 5 ml/kg) to the test animals on the basis of body weight. 2.2. Animals Albino mice weighing between 20 and 25 g were used for the analgesic studies and albino rats between 160-210 g were used for the antiinflammatory studies. These animals were of either sex and bred at the Animal House of Wellcome Pakistan Ltd. All animals were maintained on groups of five at 22 f 1“C with light/dark cycle of 12/l 2 h and with food (prepared locally) and water freely allowed. 2.3. Analgesic testing In the present study, analgesia was assessed

Ethnopharmacol.

42 (1994)

193-198

according to the method of Di Stasi et al. (1988). Mice were divided into 5 groups with each group containing 5 animals. Each mouse was held in a suitable restrainer with the whole tail extending out. An area of the tail, 2-3 cm in length, was marked and immersed in a water bath thermostatically maintained at 5 1“C. The withdrawal time of the tail from hot water (in seconds) was noted as the reaction time or tail flick latency. Control animals received 10 ml/kg of 0.9% NaCl orally while plant extracts in doses of 300, 500 and 1000 mg/kg body weight were given orally by intubation. The initial readings were taken immediately before administration of test and standard drugs and then 60, 90, 120, 150, 180 and 210 min after administration. The criterion for analgesia was a postdrug latency which was greater than two times the average predrug latency (Janssen et al., 1963). Tail flick latency difference (TFLD) or mean increase in latency after drug administration was used to measure the analgesia produced by test and standard drugs. Analgesia TFLD was calculated as follows: Analgesia TFLD = (postdrug tail flick latency) - (predrug tail flick latency). 2.4. Anti-inflammatory testing In present study, anti-inflammatory activity was determined in rats by measuring the mean increase in hind paw volume after the subplantar injection of carageenan (Winters et al., 1962). All rats were divided into five groups, each group having five rats. The animals were injected with 0.1 ml 1”/;1 carrageenan in 0.9% saline (Sigma, USA) in the right hind foot under the plantar aponeurosis (Ocete et al., 1989). The test groups were given orally 300, 500 and 1000 mg/kg of the methanolic extracts of the two plants 1 h before the carrageenan injection. The controls were given the same volume of saline, Another group of rats was treated with 300 mg/kg of acetylsalicylic acid (Aspro, Nicholas Pakistan Ltd.) orally 1 h before carageenan injection. The inflammation was quantitated in terms of millilitres using a plethysmometer (7150 Ugo Basile) immediately before carrageenan injection and then

F. Ahmad

et al. /J.

Eth~~~aco~.

42 (1994)

193-198

195

the methanolic extracts of C. pa~~cu~at~ and T. un~lata are shown in Tables 1 and 2, respectively. The methanolic extract of C. paniculatus showed a varying degree of analgesic activity at all three doses. Its analgesic activity was more potent at 500 mg/kg than at the other dose levels as indicated in Table 1. Similarly, the methanolic extract of 7’. undulata showed significant analgesic activity at all three dose levels, relative to control group (Table 1). Results indicate that the anti-inhalator activity of the methanoli~ extract of C. ~~i~lat~~ was highly significant at all three dose levels (Table 2). Whereas the methanolic extract of T.undulata failed to exert any significant inhibitory effect on carrageenan-induced paw edema in rats (Table 2), indicating little to no potential as an anti-inflammatory agent.

1, 2, 3, 4 and 5 h after carrageenan injection. The percent inhibition of edema was calculated for each group with respect to its vehicle-treated control group. The inflammatory activity was calculated by the following relationship:

AyBxloo A (Planichamy and Nagarajan, 1990) where A and B denote mean increase in paw volume of control and drug-treated animals respectively. 2.5. Statistical anaiysis Values for analgesic activity were expressed as mean increase in latency after drug administration f S.E.M. in terms of seconds whereas values for anti-inflammatory activity were expressed as mean increase in paw volume f S.E.M. in terms of millilitres. The significance of difference between means was determined by Dunnett’s t-test and values of P < 0.05 were considered significant and P < 0.01 as highly si~i~cant. All statistical procedures were performed according to the method of Alcaraz and Jimenez (1989).

4. Discussion C. panicuiatus and T.u~dulata were selected for the present study because of their ayurvedic reputation and some previously reported biological activity (Seth et al., 1963; Neogi and Bahga, 1964/1965; Bhattacharya et al., 1971; Wangoo and Bidwai, 1988). To date, however, no work had been done to test the analgesic and antiinflammatory potential of these plants. According to the present study, the methanolic extract of C.

3. Results Results of the analgesic and anti-inflammatory effects of acetylsalicylic acid (standard drug) and Table

I

Analgesiceffectsof methanolicextractsof Cefastrus Treatment

Oral dose

Mean

(mglkg)

+60

Saline control Celastrus paniculatus extract

300 500 1000

0.06 * 0.17 0.70 f 0.40 1.10 f 0.19** 0.73 l 0.34

0.39 1.38 1.68 I..50

Tecomella undulata extract

300 500 1000

Acetyl-saiicylic acid n/group = 5. *P c 0.05. *+P < 0.01.

300

panicuiutus

increase in latency

f S.E.M.

+W

and TecomeNu

unduiara

using

the mouse tail immersion

method

(s) +I20

+I50

0.31 0.16 0.22** 0.22.

0.33 * 0.29 2.1 I f 0.08** 2.39 f 0.26** 2.47 f 0.54**

0.50 2.71 3.06 3.00

0.86 i 0.24’ 0.68 f 0.1 I 0.88 f 0.24’

1.19 f 0.21 1.80 f 0.49+* I.71 f 0.28’

1.45 f 0.25. 1.81 f 0.20** 1.66 f 0.29’

2.30 f 0.32 2.12 f 0.14 3.56 f l.48**

2.76 zt 0.08** 2.96 f 0.37** 4.78 zt l.l9**

2.46 rt 0.23* 4.02 f 0.73** 3.34 f 0.83**

0.90

2.22 f 0.19*+

2.31 f 0.14’.

3.48 f 0.14**

3.84 f 0.20**

4.14 f 0.24**

l

0.15’

f f f f

+I80 f f f: f

0.22 0.22.’ 0.36’ 0.34*

0.37 3.20 3.63 3.34

+2lO zt f zt f

0.28 I .93*’ 0.37** 0.39**

0.46 4.30 4.07 2.93

(min) f f * zt

0.32 0. I3+* 0.41” O.SO**

**p

< 0.01.

n/group = 5. ‘P < 0.05.

Acetylsalicylic acid

500

300

1000

300

unduluru extract

1000

500

300

0.75 f 0.04

0‘80 f 0.01 0.80 f 0.01 0.80 l 0.02

extracts

0.93 f 0.04**

2.00 f 0.04 2.05 f 0.06 2.01 f 0.03 I.1 I f 0.07**

1.94 f 0.06 I .9-l f 0.12 1.91 f 0.06 I .os f 0.07.’

I .62 f 0.03 I .82 f 0.07 1.85 f 0.06 0.99 l 0.04**

1.24 f 0.04 1.36 f 0.05 1.36 zt 0.06

+4 h I.57 f 0.02 1.47 * 0.01** I.19 f o.oi** 1.19 I: 0.01**

+3 h 1.39 + 0.05 I.29 f 0.01** I.11 f 0.02** 1.10 f 0.05**

+2 h

(ml)

1.33 f 0.03 1.24 f 0.01** I.15 l 0.01** 0.99 f o.oO**

h

f S.E.M.

I.12 f 0.06**

1.90 f 0.10 1.94 * 0.09 I .92 f 0.06

54.80 f 1.90

-2.80 f II.10 -31.80 * II.30 -27.30 * 9.90

58.60 * 3.60 72.30 + 5.50 41.25 f 5.70

+I h

Mean

61.60 f 4.60

-2.50 f 5.20 -27.90 f 8.80 -31.90 f 6.60

(‘K)

56.70 zt 4.80

-12.50 f 6.60 -18.00 f 10.60 -9.30 t 4.50

29.10 f 2.20 57.25 f 2.70 46.90 f 5.10

+3 h

+ S.E.M.

rat paw edema

30.40 f 2.60 47.10 f 2.50 58.10 f 3.00

+2 h

inhibition

on carrageenan-induced

I.52 * 0.04 1.48 f 0.01** 1.13 It 0.02** 1.14 f 0.01**

+5 h

of Cekrstrrts ~~~~ric~r&rrtr.sand T~~~nre//u unduhrlr

I.11 l 0.04 0.97 f 0.01** 0.93 l 0.01** 0.97 f 0.01**

+I

paw volume

0.70 f 0.03 0.80 f 0.006 0.82 k 0.02 0.73 f 0.01

’

(mg/kg)

0.5

Mean

Oral dose

ePfects of merhanolic

rec~m~i~u

C&rrrUS pmiculurus extract

Saline control

Treatment

Table 2 Anti-inflammatory

58.30 f: 5.70

-4.30 f 4.70 -9.20 * 5.11 -5.55 f 2.50

23.00 f 2.00 57.22 * 2.00 47.60 f 2.60

+4 h

z B a % + B s

f 5.21 54.20 * 5.80

3 L f 8.70 -11.90

- 10.36 f 9.90 -14.30

17.20 1.30 61.69 f 1.90 49.60 f 1.90 t

+5 h

F. Ahmad et al. /J.

Ethnopharmacol.

42 ( 1994)

possessed both significant analgesic and anti-inflammatory activities (Tables 1 and 2) but the results of analgesic activity were not clearly dose dependent, i.e. the plant extract at 500 mg/kg showed more potent analgesic effect than at 1000 mg/kg This may be due to the small numbers of animals involved or to some pharmacokinetic variation. At present, the reason for this discrepancy in results is not understood and further investigations using more animals per dose should be carried out. Nevertheless C. paniculatus extract showed significant analgesic effects at all dose levels and were quite comparable with those of acetylsalicylic acid, the reference analgesic drug, both in terms of intensity and duration of analgesia (Table 1). C. paniculatus clearly exerted very significant inhibitory effects (P < 0.01) on edema induced by carrageenan at all three dose levels. While 300 mg of the extract appeared to be equally active at 300 mg of acetylsalicylic acid at +l h, it appeared to be clearly less potent than the reference drug at all other times. The oil of this plant has been used in indigenous medicine as a remedy for rheumatism and gout (Kumaraswamy and Manjunath, 1936). Triterpenoids have also been isolated from C. paniculatus and such compounds are known to possess anti-inflammatory potential (Winter et al., 1962). Thus our results support the claimed uses of this plant in folk medicine. The methanolic extract of the plant T. undulata showed significant analgesic activity, although these results were not clearly dose dependent. These analgesic effects at all three doses were generally less than those seen for acetylsalicylic acid. The earlier phytochemical studies of T. undulara showed that plant contained some sterols (Chawla et al., 1977) and such compounds can possess antiinflammatory potential (Winter et al.. 1962). Later, Taneja et al. (1975) isolated some glucosides from T. undulata which also may possess antiinflammatory activity (Della et al., 1986). However, the present study showed that the plant extract tested here clearly failed to inhibit the carrageenan-induced edema. The active component may be insufficient in quantity in the crude extract to manifest activity at the dose levels employed. paniculatus

197

193-198

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