Pharm. Méd Trad Afr. 2004. Vol.13,

Pharm. Méd. Trad. Afr. 2004. Vol.

ACllTE AND CHRONIC ANTIINFLA.MATORY PROPERTIES OF Kalanchoe crenata Andr. (Crassulaceae) EXTRACTS

Agathe Lambou FOn0², Télesphore Benoît NGUELEFACK\ Théophile DIM0^2*, Emmanuel A. ASONGALEl\'e. Pierre KAMTCHOlJlNG²

'Department of Animal Biology, Faculty of Science, University of Dschang, BP 67 Dschang, Cameroon

2Departement of Animal Biology and Physiology, Faculty of Science, University of Yaoundé 1, BP 812 Yaoundé, Cameroon

J Department of Physiological Sciences, Faculty of Medicine and Biomedical Sciences, Pharmacology and Toxicology Unit, University of Yaoundé 1 P.O. Box 8283, Yaoundé.

*corresponding author: Email: ~

Abstract

Kalanchae crenata is used in folk medicine for the treatrnent of severa! inflammatory disorders. Methylene chloride/methanol extract of K. crenata was successively exhausted in hexane, methylene chloride. ethyl acetate and n-butanol. The anti-inflarnniatory profile of the obtained extractswas investigated on carrageenan-induced paw oederna. The oral administration of n-butanol extract (600 mg/kg) produced a maximum inhibition of about 45~'o of carrageenan- induced paw oedema, The n-butanol extract also exhibited acute anti-inflammatory activity on histamine(47.51%). serotonin (54.51%) and forrnalin (3597%) induced paw oedema. In the chronic inflammation, this extract showed signiftcant maximum inhibition of 58.33°ô on the ninth day of treatment. The ulcerogenic assay shows that ulcer indices afterp. (J. treatrnent with n-butanol extract were 0.0±00 at 300 mg/kg and 0.4± 0.2 at 600 mg/kg. On the basis of these findings, it may be inferred thatK. creuatais an anti-inflammatory and anti-arthritic agent which blocks the cyclooxygenase and histamine pathways The results are in agreement with the traditional use of the plant in inflammatory conditions.

Key words: carrageenan. histamine, 5-HT, formaI in, anti-inflammatory activity, Kalanchoe crenata

1-1ntroduction

Kalanchoe l'reno/a Andr. (Crassulaceae) commonly known as "never die" or "Dogs livet" has been traditionally used for the treatment of many ailments such as earache, srnall pox, headaches. inflammation. pain, asthrna, palpitations, convulsion and general debilitv. Aqueous and alcoholic extracts ofK. crenuto's leaves contain alkaloids and saponins (Sofowora, 1993)

ln early studies. aqueous and ethanolic extracts ofK. crenata was found to inhibit pain induced by acetic acid, forrnalin hot plate and pressure (Nguelefackelal, 2004) This work was

Pharm. Méd. Trad. Afr. 2004. VoU3, pp.57-66

aimed at providing scientific validation of the clairned ethnopharrnacological properties, bv investigating anti-inflamrnatory properties of the leaves extracts

2-Materials and methods 2- /-Plant materials

K.crenatas leaves were collected in Dschang City (West Cameroon) in October 2002.

and authentified by comparison with a voucher specimen number 50103IYA in the National Herbarium, Yaounde. Cameroon. 2 kg of the air dried leaves were ground to a fine powder and extracted with methylene chloride/methanol (CH2Cb/CH,OH) for 3 days (72 h) The resulting extract was concentrated in vacuum over rotavapor to obtain 234.7g ofCH2CI 2/ CH10H extract.

212.4 g of this extract were exhausted successively with hexane, methylene chloride, ethyl acetate and n-butanol The following fractions were obtained: hexane (92.2 g); CH2C1 1(4.7 g);

ethylacetate (7.1 g): n-butanol (37 g) and aqueous residue (235 g). CH2CI 2/MeOH. CH2C1 2and n-butanol extracts were dissolved in 2.5~·o DMSO and 2.5~..oTween 20; while hexane and ethyl acetate extracts were dissolved in 3% DMSO before giving orally at the doses of 300 and 600 mg/kg of the animals body weight.

2-2. Phytochemical screening

The total extract as weil as its fractions were submitted to the test of Liberman Buchard, the ferric chloride test, the copo of magnesium test and the Vanillin-sulphuric acid test in the goal to determine the presence of sterols. phenolic cornpounds, flavonoids and saponins respectively

2-2 Chemicals

Indomethacin (Sigma), pyrilamine Maleate (Sigma). diclofenac, carrageenan (Sigma), Histamine (Fluka), 5-hydroxytryptomine (S-HT) (Fluka) and formaldehyde (Roth) were used.

OlmI of oederna-inducing agents was administered into the plantar surface of the right hind paw of the animaIs.

2-3 Animais

Male and female Wistar rats (140-190 g) were used for the investigations. They were obtained from the Animal House, Department of Animal Biology and Physiology, University of Yaoundé 1 Cameroon The animaIs were divided into groups of five and fasted 24 h before the experimern.

2--/ Pharmacological studies

2--/-/ Carrageenan-inducedpawoedema

The extracts (experimental groups) a control vehicle (solution contain 2.5% DMSO and 2.5% tween 20) (control group) and indornethacin 10mglkg (reference group) were given oraJly one hour before subplantar administration of 1% carrageenan suspension in 0.9% NaCI The volumes of the injected paws were measured 30. 60, 120, 180. 240, 300 and 360 min after

58

Pharm. Méd. Trad. Afr. 2004. Vol.13,

induction ofinflammation using Ugo Basile Plethysmometer N" 7! 50 as described bv winter el

al. (1^962). The oederna was expressed as an increase in paw volume due ta carrageenan injection

2--1-/ Histamin and Serotonin-inducedpaH' oedema

ln another set of experiment serotonin and histamine were used as phlogogen agents The n-butanol extract ofK. creuata(experimenta! groups) and control vehicle (solution contain 25%

DMSO and 2.5 % Tween 20) were administered one hour before injection of intlammatory mediators

The respective strength of oedemogens, the volume injecred and the time of determination of oederna volumes are indicated in parcntheses ; serotonin (10.¹g/ml, Olmi. 30 min) : histamine ^(10.1 g/ml, 0,1 ml, 60 min) (Singh I!I al, 1996). The reference groups were treated with indomethacin (10 mg/kg) or pyrilamine rnaleate (1 mg/kg) respectively.

The oedema volume was determined as mention previously.

2--1--1. Formalin-inducedpail ocdctna

The inflammation was produced by subaponevrotic injection of 0.1 ml of 2°/0 forrnaldehydeone hour after oral administration of n-butanol extract ofK. creuata.diclofenac(5 mg/kg) or control vehicJe (solution contain 2.5% DMSO and 2.5% tween 20) The oederna volume was determined 1. 2 and 4 hours after injection of formaldehyde.

The same animais were further treated with the extract or diclofenac tor the following 9 days A second injection of formaldehyde was done on the third day (Hosseinzadeh and Younesi, 2002) The dai/y changes in paw size were measured plethvsmographically.

2--1--1.1llcerogenic activtty

Two hours after test drugs administration {n-butanol extract ofK.crenata, indomethacin ( 10 mg/kg) or control vehicle), animais were sacrified by cervical dislocation, stomach isolated and opened along the greater curvature. Ulcerated surface was measured and scores were attributed according to the table described byMartinelal. (1993J.

2--1-5. Statistical analysis

Ail values were presented as mean values ± S.E.M from fivc rats. The statistical significance between the treated groups and the control group was calculated through the analysis of variance (ANOVA) followed by the Students "t" test. P values less than 00:;

(P<OOS)were considered significant.

3-Results

Pharm. Méd Trad Afr. 2004. Vol.13,

erhvl acetate fractions. Flavonoids and saponins have been detected inCH¹0 H/CH2(ï èextract and its n-butanol fraction

3-2. ('arrageenan inducedpmi!oedema

The effect of the K. crettata 'sextracts on carrageenan induced-paw oedema is shown in table 1. The CH2Cl 2/MeOH, ethyl acetate extracts and aqueous residue showed at 600 mg/kg a maximum anti-oedernatous effect of about 41.75; 40.10 andJ571~'o respectively one hour afler carrageenan administration. The hexane, CH2Ch and n-butanol extracts showed a maximum anti-inflarnrnatory etTect ofabout44.34 (30 min),39.01 (1 h) and45.80%(1 h) after carrageenan administration respectively. This effect was maintained up to 25.16 and 30% for the hexane, CH2Chand n-butanol extracts respectively.

The anti-inflammatory effect induced by indomethacin progressively increased and reached a maximum (6591%) at 3 h, and the effect was maintained up to 6 hours.

3-3. Serotonin and histamine inducedpOli'oedema

Table 2 shows the effect ofthe n-butanol extract ofK. crcuataon serotonin and histamine induced paw oederna. At the dose of300 mg/kg, the n-butanol extract ofK. Crenatasignificantly reduced the paw oedema formation induced by histamine (41.98%).The percent age inhibition at the dose0[600 mg/kg was 54.51 and 4751% on serotonin and histamine induced inflammation respectively. Pyrilamine maleate at the dose of l mg/kg inhibited histamine-induced inflammation by 58.01% while indomethacin exhibited a percent inhibition of 48.49% on inflammation inducedby serotonin.

3--1. Formalininduced pail' oedema

The n-butanol extract of K. crenata (300 and 600 mg/kg) and diclofenac (5 mg/kg) produced significant inhibition of forrnalin induced inflammation by 32.57, 35.97 and 43.62%

respectively 4 h after formalin administration (table 3).

ln the chronic inflammation, the extract showed significant inhibition On the seventh day of experiment. the inflammation of the paw of animais treated with extract (600 mg/kg) and diclofenac⁽⁵ mg/kg) was3140and3107% respectively, showing an inhibition of 5594%. An inflammation of 21.37% was observed in rats treated with extract (600 mg/kg) on the ninth day, presenting a maximum inhibition 01'58.33%

3-5. Ulcerogenic activitv

As shown in table 4, 300 mg/kg of the n-butanol extract ofK. crenata fail to induce gastric ulceration. At the dose of600 mg/kg, 2 over 5 rats treated show blood vessels dilatation, corresponding to general ulcer index of 0.4. Ali the animais that received indornerhacin presented significant ulceration surface of6.85 mrrr'. lndomethacin as weIl as n-buranol exrract (300 mg/kg) significant reduced mucus weight.

60

Pharm. Méd. Trad. Afr. 2004. Vo!.13, pp.57-66

4. Discussion

The experimental data of the present study indicates that K. crcnata extracts possesses significant anti-inflarnmatory activities on various phlogistic agents.

Carrageenan induced intlammation involved three distinct phases of mediators release including serotonin and histamine in first phase, kinins in second phase and prostaglandin in third phase (Singh etal.. 1996) CHzCh/iVfeOH extract of K crenata and aqueous residue have significantly inhibited carrageenan induced paw oedema only in the first phase, suggesting an inhibitory effect on the release of histamine and/or serotonin Anti-inflamrnatory effect of the ethyl acetate extract was prolonged to the third hour, suggesting an inhibition of the release of kinins. Hexane, CHzClz and n-butanol extracts showed a significant inhibition of the oedema on the three phases. This oedematous response was also significantly reduced in rats pre-treated with indomethacin, compound known to be cyclooxygenase inhibitors. According to Chawla el al.(1987) and Dongrno el al. (2001), 5-lipoxygenase inhibitors also possess anti-inflarnmatory activity on carrageenan-induced oederna The inhibirory effect of the hexane and n-butanol extract on the third phase of the oedema could be due to an inhibition of 5-lipoxygenase and/or cyclooxygenase, both enzymes involved in the formation of 1eukotrienes and prostaglandins respectivelv The n-butanol extract was more active than others extracts and was chosen for further studies. Results obtained with n-butanol extract on the carrageenan-induced oedema were confirmed on the acute inflammation induced by formalin. According to Yuh-Fung etal. (1995), acute inflammation induced by formalin results from cell damage which provoked the production of endogenous mediators such as histamine, serotonin, prostaglandins and bradykinin. Formalin-induced oedema was significantly inhibited by n-butanol extract of K.

crenata.

ln order to ascertain the effect of the extract on mediators' activities, the extract was tested against inflammation induced by histamine and serotonine. lt has been observed that n- butanol extract ofK creuata inhibited histamine and serotonin-induced oederna. The inhibitory effect ofK. crenata inferred that the extract mal' be acting like pyrilamine as an antagonist of these mediators which also significantly inhibited inflammation induced by histamine

n-butanol extract ofK. crenata was further essayed on chronic inflammation induced bv formalin.It is weil known that inhibition of formalin-induced pedal oederna in rats is one of the most suitable test procedures to screen anti-arthritic and anti-inflammatory agents as it closely resembles human arthritis (Greenwald, 1991). Formalin-induced arthritis is a model used for the evaluation of an agent with probable anti-proliferative activity. This experiment is associated with the proliferative phase of inflammation (Banerjee el al., 2000) Since n-hutanol extract ofK.

creuata could significantly inhibit this mode! of inflammation. it can be though that it possesses

"

Pharm. Méd. Trad. Afr. 2004. vot.ts, pp.57-66

anti-proliferative and anti-arthritic activities which have been observed with diclofenac. a Icyc)ooxygenase inhibitor

Since results from the present study strongly indicate the non steroidal anti-inflammatory like activity of the extract, its ulcerogenic effect was tested on starved animais Non steroidal anti-inflamrnatory drugs are thought to impair the rnucosal defence properties of the stomach and intestine. They act by inhibition of cyclooxygenase, therefore inhibiting the production of gastric prostaglandins which leads to reduction of gastric mucus production and an increase in mucosa\

permeability (Jain et al.. 2002). According to Kryvola el al. (2003), anti-histaminic agents protect the gastric mucosa against ulceration by diminishing gastric acid production. The n- butanol extract ofK.crenatawhich reduced the mucus weight but induced little ulceration of the gastric mucosa could act by dual inhibition of cyclo-oxygenase and histamine.

The presence of flavonoids in the n-butanol extract ofK.L'relia/a may account for its anti- inflarnmatory activity. Many compound From this class have been found to exhibit anti- inflarnrnatorveffects (Martini et al., :2004;Tokerc:/al ..2004)

ln conclusion, the results show the marked and dose-related anti-inflarnrnatory and anti- arthritic activities of theK. L'ret/a/aextracts. These results corroborated the traditional use of the plant in inflarnmatorv conditions Their active principles specifically inhibit certain chemicals- induced oederna in rats.

References

J - Banerjee, S., Sur, T K, MandaI, S, Chandra Das, P., Sikdar, S.(2000). Assessment of the anti-inflarumatory effects of Swertia chirata in acute and chronic experimentalmodels in male albino rats.ludiun Jourual(~rPharmacology 32,21-24

2- Chawla, A. S., Singh, M., Murthy, M. S, Gupta, M P, and Singh, H. (1987). Anti- intlammatory action of ferulic acid and its esters in carrageenan-induced paw oedema mode!. Indian Journal

(i

3- Dongrno, A. B., Karnanyi, A., Anchang, M. S, Chungag-Anye Nkeh, B., Njarnen. D., Nguelefack, T B., Nole. 1., Wagner H. (2001) Anti-intlammatory and analgesie properties of the stem bark extract of /~/y/hrophlel/m suaveolens (Caesalpiniaceae), Guillemina Perrottet.Journal oflrhnopharmacology 7(2-3), 137-141

4- Greenwald RA (1991) Animal models for evaluation of arthritic drugs Meth t-ind Cli«

Pharmacol Vs,75-83.

5- Hosseinzadeh, H. and Younesi, H (2002). Antinociceptive and anti-inflarnrnatory effects of Crocus sativusL. stigma and petai extracts in rnice.HM!' Pharmacology2,1-8

6- Krylova, S.G, Razina, T. G., Zueva, E. P, Amosova, E. N.. Shilova, N. V, Dugina, Y L.

and Epstein. 0 1. (2003) Analgesie and anti-inflamrnatory activity of antibodies to

Pharm. Méd Trad Afr. 2004. VoU3,

histamine under experimental conditions Hnlletm oilxperimental Ifi%g\'UIIUMedicine, supplement 1.83-34

7- Iain, N K, Kulkami, S K, Singh, A (2002) Modulation ofNSAID-induced antinociceptive and anti-inflarnrnatory effects bya2 adrenoceptor agonists with gastro protective effects.

I.fe Sciences 70. 2857-2869

8- Martin. 1\1. J, Motilva, V and Alarcon De La Lastra (1993) Quercetin and Naringenin Effecton u1cerformationand gastric secretion in the rat. PhytotherapyRescarch, 7. 150- 153

9- Martini. N. D. Katerere, D. R. P and EJoff J N (2004) Biological acnvrty of five antibacterial flavonoids from Combretum erythrophylltttn (Cornbretaceae). .lottrnal of

j-j/1J/ojJhm'l1loc%j!;Y93, 207-212

10- Nguelefack, T B. Fotio, LA, Watcho, P.,Wansi, S. Dimo ,T, Kamanyi, A (2004) Analgesie activities ofaqueous and ethanolic extracts of the leaves of Kalauchoe cretuua (Crassulaceae) PhytotherupvResearch 18.385-388

11- Singh, S. Majurndar, D. K, Rehan H M. S (1996) Evaluation of anti-intlammatory potential of tixed oil of Ocimum sanctum (Holybasil) and its possible mechanisrn of action . Journal ofEthnopharmacologie 54, 19-26

12- Singh, GB. Bani. S, Singh, S, Bane Rjee S. K (1997) Anti-inflammatorv actions of Euphorbiasplendens extracts. Phytotherupy Research 1L 76-78

13- Sofowora, A (1993). Medicinal plants and traditional medicine in Africa. 2"^d ed polygraphic ventures. LTD Ibadan pp 207-209

14- Toker, G, Kupeli, E.. Mernisoglu. M, Yesilada E, (2004). Flavonoids with antinociceptive andanti-inflamrnatory activities from the leaves ofTrùa wgellleu(silverlinden) Journal ofl-rhnopharmacology95. 393-397

15- Winter, C. A, Risley,E. A and Nuss, G. W.(1962) Carrageenan-induced oedema in hind paw of rat as an assay for anti-inflarnmatory drugs. Proceediug» oi' societv fi)"

l-xperimental HIO/oK)' m/LI Medicine 11 l, 544-547

16- Yuh-Fung,

c.,

Table 1: Effcct ofK.crenaui cxtracts on carragccnan induccd pa" ocdcma in rats Doses

5il 611

Jh 4h 2h

- p()lii-celllage,; dtuhibit1 < 1 1 \ - - ' - - - -

--.L.----;-__

1~llmin Ih

~h ~h 5h (,h

luflunuuations(!',VL11mIt 2h

- - - _^{. .}----:-:---111

(rn~lkt!1 30min 'l'rai telllellls

0.00 Il.(11)

0.(1) O,llI) IUiO

. _ - - _ . _ - - - - _ . _ - -0.00 O.()O

(I.(,(l±(J.(I~

O.6-1±(l.fJ~

0.7010II.II.~

IU'h0.02 OHl±(J.O.~

IU61oO.Dl (J2.~±n.zz

Coutrul /1

CII,Cl,ICIhOB ~IiO O.2l\±no~ IU7±0.04 07210 0.0(, OX(~±00'1 0.(,7±(lOR 072±o.o« 0.77± OOt) -25.21 -3.2') -X.OK ·1.00 ~.2!1 -11.1/3 -If,l(,

*p<O.OS. **p<O.O 1 comparcd with control

O.SII±IlIl' 075±IUl4 1221>1) -lOlO ,,8.<>2 2~.XI Il.71l±O.OI~ n.(,)1oo.1O 140.00 ~17) 2.~.<j5 6(JI

o.5ü o.1l5 OA2± (L05* 1 17..l9 J')OI 26.1).4 17.5~

"I:j

f ~

~

~l::l

~

~

at-v

a~

~:-- ...

~w

~

';'-l 0-0- 'ilk

~2.21 5 X6 3271

5401 :'~~,) -.~4.25 ·1 (,1 l,

) I~ -1.5~ (,2S

5.1~ -S..'3 R.i>l~

Il.110 ·23.71' ·12

rt

IfJ.2X 16..1) .~().22

lOlO 308)

-li.57 -1111 -7~X

57.~2

2h.no 25 ')2 .~X~2

21.71 197) 2~55

l3.~2 5X<> ulX

-22.2X 1.25

lUX

.~l)._'-f

ilS') 1 -.'.59

2(;.'14

~:'.Xll

57.7'1, -7.6') --l2X5

35.71 -IX.l)1) 31.30 3').1J

().~X±0.0.' O.~I±0.02 1 -1-1.'-1 27.~7 2'J.'1~ 32.X~

0,(. I±(l.O~ 1).f)I~n.o« ¹ -9.5f,

0.6:'±O.()~ n.!>.?: ()02 1 !If,,! ~()70 15.:'6 1(,7'1

O(,I± 0.(J7 Of,m n 0(, ¹ Id) 25.27 5 ..~8 21H1) 0.71± 0.06 O.7(~0.0(, 1-21.7.' --I.3l) _11'17 -<J.77

lI~-,1o (I.IJ~. li~X1o()o(,·· 1 2'-l..l<j

O.52±O(J.1* ():'(~0.05·· i -2:121 7.(,') 1~.l)7 17.0--1

0.?X± II.IJ:' IU;:,± 0112 0.87±(J.O~ O.77± Il.02 Il.17± 11.07·· Il.2'1± 0 ..1l7·· U2'i± 0.04^u U.3Il± 1/.05··

02R±O.fn O.3.1±001 11.:'()±1i.0~ Oc>()±D.Il--I" 0.541oDU2 ....

o l '-J±O.OI O.22± OO'-l· Il.~<J±l!.ll:'· Il (.:'±li07 Il.5R± 0 07 lI.21±1l.0~ ^[1.l')10lI.O~u lI.Sfl±OO~ ON,±. O.Où O.M,± 0 03

Il.21± 0.02 o.ntIl.O-I 0 f,l±(J.D~ 0I).~±lI.1l5" lI:,l± IJ()(\·

0.27± 1J.(l4 [13-1± 0.D3 lI.7-l± Il.lIÙ (Un1oo.o« IJ.7.:!±. O.M

D.17± 0.02 0.20± D.02·· 1l..'ll± 011-1" [1~X± O.{J.~·· (I.-IX:!.(J.o~'"

I/.I:'± lU» II 1H±o.o.~n 02X±(Ulh"

0.25± lI.02 tU')±()(J.~ 0 (,')± 0.05 (J.70± 0 03 O.6.H D.05 O.17±o.m O.2± 10.03·" (J·II± (JlI7· O.Wttl.02*· (J7~±1!.l15 D.12101I.01· 1)2(,±Il liS 0~c>±Il.02'' O.S.\±.0.lI4'" 0.51± 002'"

OU±IJ.(J2" 021100.02"* 0.50±o.OX 0.7:'±OIO O.()(,1o{J.IO

O.l~±0.(J 1* (J.2~±0.01" OAS±lun

Jil 300

.~Oll .~t~)

iloo

(,(10

(,(JO

.~(J(I

600 .100

(,(JO (,Of)

residuc

~'lrarl

cxtrurt exuuct

AqUl:OUS

n-butauol f:t1l\lm:elille

hulomcthaci»

CII,Chc'.\lnlcl 11",;\lIe cxtruct

01+:>-

Pharm. Méd. Trad. Afr. 2004. Vol.13, pp.

Table 2. Effcctofn-butanol cxtractofK crenataon histamineandscrotonin induced paw ocdcma

- - - - ----_._--_._...^_--~_._-_..._---~---_.--_._----_.._----".". -

Group Dose(p.o) Meanocdcrna volume (ml) Pcrccntagcinhibition - - - _^..

Histamine Serotonin Histamine Serotonin Control 10ml/kg

o

n-butanol 300 mg/kg 0.21 ±0.04* OAI ±0.02 4198 2140

extract 600 mg/kg 019±OOI** 0.24±(JO/** 47.51. 54.51

Pyrilamine 1mg/kg

o

lndomethacin 10mg/kg 027±002** 4849

*p<O.O:'. **p<O.O1 comparcd with control

TablcJ .Effcct of n-butanol ofK.crenataonforrnalin induccd pa« ocdema

._----.---_._-.~--_._---_._--.._._--_.,---

Dose Mean oedema volume (ml) Percent inhibition

Group .__. _ - - - - _ .--_.

__

(p.Cl) Ih 2h 4h lh 2h 4h

-~-_..- - - -

Control 10 ml/kg O.'i2±0.04 063 ± ()O6 O.70±OO6

n-butanol 300 mg/kg OAI±00-1- OAl)± (l.O-l- 0.-1-7±O.Oô** 1992 2158 32.57 extract 600 mu/ku_{o ~} 035±0.03* (UX±00'+** OA2±()O5** 3295 3L).36 35.97 Diclofenac 5 mg/kg 0.32+0.02** OA3 ± 001 * 03lJ±002** 37.93 3047 43.62

*p<005. **p<O.O1 comparcd with control

Pharm. Méd. Trad. Afr. 2004. VoU

120

100

~

;:R0 80 c0

~ 60

EE

CIl 40

<;;::

C

20

0

dl d2 d3 d4 d5 d6 d7 dB ^d9

1 1 1

--<>-control - - n-butaro 1300

- ·6· - n-butarol600 _ _ _ _ diclo fenac

d10 Time (day)

Figure 1. Effect of n-butanol extract of K. crenata on chronie inflammation induced by formalin *p<()(J:ï. **p<lJ.()l. ***p<()()(J 1cornparcdwith control

Table 4. Ulcerogcnic activitics of n-butanol cxtract ofK crcnata

Doses Mucus wcight Ulccnucd Ulceration ulccratcd ulccratcd

Drugs (mg/Kg) ( mg) surface(IIUU') Indice surface('Yu) animais«~.,.)

Control (.2'>X ±~sn 0.0±00 0.0±0.0 0 0

n-butanol WO ~(j'x,±1.6~* oo±oo 0.0±00 0 0

cxtract

Blood vcsscl s

(,00 _~5.32±(lO') O~±0.2 ⁽⁾ ~O

dilation

indomcthacin 10 _~O.S()± 3.5')** (,.X5±^1.')X** 2.66±O.l(~*"'* tU\! JOO ---

*p<O.OS, **p<O.O 1, ***p<O.OO 1 eompared with eor.tro1

66