Acute and chronic antiinflammatory profile of the ivy plant, Hedera helix, in rats.
Hedera helix is a plant well-known as ivy or English ivy, and a member of the Araliaceae family. In the present study, we tested the possible antiinflammatory effects of a crude saponin extract (CSE) and a saponin's purified extracts (SPE) of Hedera helix in carrageenan- and cotton-pellet-induced acute and chronic inflammation models in rats. Both the CSE and SPE of Hedera helix were found to have antiinflammatory effects. The most potent drug screened was indomethacin (89.2% acute antiinflammatory effect), while the most potent extract screened was the CSE of Hedera helix at 100 and 200 mg/kg body wt. doses with 77% acute antiinflammatory effects. For testing chronic antiinflammatory (antiproliferative) effects, the cotton-pellet-granuloma test was conducted. Indomethacin was found to be the most potent drug in the chronic phase of inflammation, with 66% effect. The SPE of Hedera helix was more potent than the CSE in its chronic antiinflammatory effect (60% and 49%, respectively).
Key words: Inflammation, antiinflammatory, Hedera helix, ivy plant, carrageenan, cotton pellet, rats
Inflammatory diseases are treated currently with steroidal and nonsteroidal antiinflammatory drugs (NSAIDs). NSAIDs exert their effects by inhibiting the metabolism of arachidonic acid by both cyclooxygenase and lipoxygenase enzyme pathways (Insel, 1996). Despite their widespread use, NSAIDs are often associated with severe adverse effects, the most common being gastrointestinal bleeding (Fung and Kirschenbaum, 1999). Because of these effects, safer compounds are needed.
Hedera helix is well-known as ivy or English ivy, and is a member of the Araliaceae family. The fresh leaves and fruits are toxic, causing gastrointestinal irritation, bloody diarrhea and death (Baytop, 1984). Its best-known effect is to cause contact dermatitis, and several cases have been reported (Garcia et al. 1995, Massmanian et al. 1988, Hausen et al. 1987). Additionally, antibacterial (Cioaca et al. 1978), antihelmintic (Julien et al. 1985), leishmanicidic (Majester-Savornin et al. 1991), in-vitro antispazmodic (Trute et al. 1997) and antifungal (Moulin-Traffort et al. 1998) effects of Hedera helix extracts have been reported.
To investigate the effects of drugs on the acute phase of inflammation, models induced by pro-inflammatory agents such as carrageenan, dextrane, formaldehyde, serotonin histamine and bradykinin in rat paws are employed (Campos et al. 1995). Chronic inflammation models produced by implanting a foreign body under the skin are used to study the effects of a drug on the proliferation phase of inflammation (Gilman et al. 1985, Suleyman et al. 1999). Carrageenan, a mucopolysaccaride, is perhaps the most commonly-used and well-studied of these phlogistics (Leme et al. 1973), producing a maximal edematous in 3 h. While the carrageenan model is typically associated with activation of the cyclo-oxygenase pathway and is sensitive to glucocorticoids and prostaglandin synthesis antagonists, the early phase of the carrageenan response is due to the release of serotonin and histamine (DiRosa et al. 1971).
In the present study, we tested the possible antiinflammatory effects of the crude saponin extract (CSE) and the saponin's purified extracts (SPE) of Hedera helix in the carrageenan- and cotton-pellet-induced acute and chronic inflammation models in rats.
* Materials and Methods
The leaves of Hedera helix were collected in Marseille, France, in September 1999. The material was identified by Riad Elias, a staff member of Laboratory of Pharmacognosy and Homeopathy, Pharmacy Faculty of the Mediterranean University, Marseille, France.
Hedera helix extract
The materials were shade-dried. One kg of crushed plant leaves were extracted three times with seven liters of 80% EtOH. After evaporation of the organic solvent, the aqueuos phase was treated with one liter of CH[Cl.sub.3] four times, and then with one liter of BuOH three times. The BuOH was evaporated and the residue was dried in a vacuum oven at 50[degrees]C. The yield of CSE was 100 g for Hedera helix. The obtained CSE was dissolved in 400 ml of MeOH and precipitated in 2 liters of acetone. After filtration, the residue was dried in a vacuum oven at 50[degrees]C. The yield of SPE was 80 g for Hedera helix. Both extracts contain mono- and bidesmoside triterpene glycoside derivatives of oleanolic acid and hederagenin, as described previously (Elias et al. 1991).
In this study, 66 adult male Wistar albino rats weighing 180-210 g each, obtained from Ataturk University, Faculty of Medicine, Department of Pharmacology Experimental Animal Laboratory, were used. The rats were fed standard laboratory chow and tap water before the experiment. The animal laboratory was equipped with automatic temperature (22 [+ or -] 1[degrees]C) and lighting controls (14 hours light/10 hours dark). Rats were divided into groups, each containing 6 individuals and each of the groups were kept in different cages. The ethical guidelines for investigations using conscious animals were obeyed and the procedures were approved by the University ethics committee.
Antiinflammatory effects of CSE and SPE of Hedera helix were investigated in an aseptic arthritis model, which was induced by carrageenan and cotton-pellet-granuloma tests in subsequent doses. The ratio of the antiinflammatory effects of Hedera helix extracts were calculated using the following equation: Antiinflammatory activity (%) = (1 - D/C) * 100 where D represents the percentage difference in paw volume after the compounds were administered to the rats, and C represents the percentage volume difference in the control group (Suleyman et al. 1999).
Carrageenan-induced paw edema in rats
Hedera helix CSE and SPE in 50, 100 and 200 mg/kg body wt. doses, and indomethacin 20 mg/kg body wt. doses were given to rats orally by feeding tube once daily for 4 days. Two h after final administration of the compounds, 0.1 ml (1%, w/v) carrageenan solution in distilled water was injected subcutaneously into the plantar surface of the right hind paw. The paw volume was measured using a plethysmometer: before injection and 5 times at 1-h intervals (Birch et al. 1992). The antiinflammatory activity in animals receiving Hedera helix extracts was compared with that in indomethacin and control groups.
In this series, the effects of Hedera helix extracts and indomethacin on the proliferation phase of inflammation were studied using a cotton-pellet-granuloma test. Doses of 100 mg/kg body wt. CSE; and 100 mg/kg body wt. SPE of Hedera helix, and of 20 mg/kg body wt. indomethacin were administered orally to 3 groups separately. The same volume of distilled water (1 ml) was administered to the control group. After 30 min, the animals were anesthetized with 25 mg/kg thiopenthal sodium, intraperitoneally. Under sterile conditions, cotton pellets, weighing 7 mg each, were implanted an interscapular distance under the skin. The same doses of Hedera helix extracts and indomethacin were administered once a day for a period of 7 days. The rats were killed by a higher dose of thiopenthal sodium on the eighth day, and the pellets surrounded by granuloma tissues were dissected out. The moist pellets were weighed and then dried at 70[degrees]C, after which, they were weighed again and the antiproliferative effect of Hedera helix extracts compared with the control and indomethacin groups.
Values are presented as mean [+ or -] SEM. Independent samples-t test and analysis of variance (ANOVA, Dunnet method) were used for the evaluation of data and p < 0.05 was accepted as statistically significant.
Despite the fact that they have weaker antiinflammatory effects than indomethacin, both CSE and SPE of Hedera helix were found to have antiinflammatory effects. For the acute phase of inflammation, carrageenan-induced paw-volume increase, and the effects of the indomethacin and Hedera helix extracts were evaluated. Because of the most potent effect was seen in the fourth hour, prior to others, statistical data for that period was regarded to evaluate acute antiinflammatory effect. The most potent drug was found to be indomethacin (89.2% acute antiinflammatory effect) while the most potent extract was found to be CSE of Hedera helix at doses of 100 and 200 mg/kg body wt., with 77% acute antiinflammatory effects. The same drug in a dose of 50 mg/kg body wt. exerted a 51% antiinflammatory effect. For the SPE of Hedera helix, acute antiinflammatory effects of 41.5% for 50, 60% for 100 mg/kg doses and 67.7% for 200 mg/kg doses were found. For further information, please refer to Table 1.
For testing chronic antiinflammatory (antiproliferative) effects, the cotton-pellet-granuloma test was conducted. Indomethacin was found to be the most potent drug in the chronic phase of inflammation, with 66% effect. The SPE of Hedera helix was found to be more potent than the CSE in terms of chronic antiinflammatory effect, at 60% and 49%, respectively. For further information, please refer to Fig. 1.
[FIGURE 1 OMITTED]
Triterpene saponins of Hedera helix have already been tested in several applications. Buddlejasaponin's and saikosaponin's in-vivo antiinflammatory effects on mouse ear edema (Bermejo-Benito et al. 1998) and zanhasaponin's acute and chronic antiinflammatory effects in different models (Cuellar et al. 1997) have been reported. For the first time in the literature, we have demonstrated the antiinflammatory effects of Hedera helix due to mono and bidesmoside triterpene glycosides. Despite the fact that they have weaker antiinflammatory potential than indomethacin, in all of the groups tested, we found quite effective antiinflammatory action. The CSE of Hedera helix was more potent than the SPE form in the acute phase, while the SPE form showed better results than the CSE form of Hedera helix in the chronic phase of inflammation.
It was reported that the antiinflammatory effects of several agents result in the partial inhibition of inflammation-mediator release (Amadio et al. 1993). Subcutaneous injection of carrageenan into the rat paw produces plasma extravazation (Szolcsanyi et al. 1998), and inflammation characterized by increased tissue water and plasma protein exudation with neutrophil extravazation and metabolism of arachidonic acid by both cyclooxygenase and lipoxygenase enzyme pathways (Gamache et al. 1986). There are biphasic effects in carrageenan-induced edema. The first phase begins immediately after injection and diminishes in 1 h. The second phase begins at 1 h and remains through 3 h (Garcia-Pastor et al. 1999). It is suggested that the early hyperemia of carrageenan-induced edema results from the release of histamine and serotonin (Kulkami et al. 1986). Both Hedera helix extracts and indomethacin started to block inflammation in the first measurement. So it seems possible that Hedera helix blocks histamine and/or serotonin release in the first phase of acute inflammation. On the other hand, the delayed phase of carrageenan-induced edema results mainly from the potentiating effect of prostaglandins on mediator release, especially of bradykinin. Hydrocortisone and some NSAIDs inhibit strongly the second phase of carrageenan-induced edema, but some others are effective against both phases (Kulkami et al. 1986). In the light of these data and Table 1, Hedera helix extracts seem more effective in the first phase of acute inflammation than in the second phase. Therefore, Hedera helix extracts may block histamin and/or serotonin release better than the prostaglandin and/or bradykinin.
It was also observed that both extracts of Hedera helix are effective in chronic inflammation, despite the fact that the effect was not strong as indomethacin. Chronic inflammation is a reaction arising when the acute response is insufficent to eliminate proinflammatory agents, and induces a proliferation of fibroblasts and the infiltration of neutrophils and exudation (Dunne, 1990). Chronic inflammation occurs by means of the development of proliferative cells. NSAIDs cause a decrease in granuloma tissue arising as a result of cellular reaction, released by inhibiting granulocyte infiltration to the foreign body implanted (Ionac et al. 1996). According to the chronic inflammation model of the cotton pellet, the Hedera helix SPE and CSE may exert their effects by inhibiting the functions of macrophages and fibrosis.
The results of the present study show that Hedera helix CSE and SPE are potent inhibitors of acute and chronic inflammation. The mechanism of the effect may depend on inhibition of the formation of several inflammation mediators. Detailed studies are needed to clarify the mechanism(s) of the antiinflammatory effects of Hedera helix triterpene saponins.
Table 1. The effects of CSE (Crude saponin extract) and SPE (Saponin's Purified Extract) of Hedera helix in subsequent doses and indomethacin (Indomet.) in carrageenan-induced acute paw edema. Right paw volume (ml x 100) Drugs Dose n Before 1 h mg/kg body wt. Vehicle -- 6 77 [+ or -] 2.5 104 [+ or -] 6.4 * Indomet. 20 6 101 [+ or -] 2.9 112 [+ or -] 3 * CSE 50 6 112 [+ or -] 1.6 113 [+ or -] 1.4 CSE 100 6 117 [+ or -] 3.2 128 [+ or -] 3.2 * CSE 200 6 113 [+ or -] 3.1 113 [+ or -] 3.6 SPE 50 6 110 [+ or -] 1.3 115 [+ or -] 2.5 SPE 100 6 115 [+ or -] 4 123 [+ or -] 3 SPE 200 6 113 [+ or -] 1.3 130 [+ or -] 3.6 *** Right paw volume (ml x 100) Drugs 2 h 3 h Vehicle 134 [+ or -] 2.1 **** 139 [+ or -] 2.7 **** Indomet. 110 [+ or -] 2.4 * 109 [+ or -] 2.1 CSE 129 [+ or -] 4.5 * 142 [+ or -] 3.1 **** CSE 130 [+ or -] 4.5 * 131 [+ or -] 4.4 * CSE 119 [+ or -] 4.4 126 [+ or -] 4 * SPE 150 [+ or -] 3.2 **** 149 [+ or -] 4.4 **** SPE 137 [+ or -] 3.3 *** 139 [+ or -] 3 **** SPE 147 [+ or -] 4.8 **** 136 [+ or -] 2.4 **** Right paw volume (ml x 100) Drugs 4 h 5 h Vehicle 142 [+ or -] 1.7 **** 140 [+ or -] 1.6 **** Indomet. 108 [+ or -] 4.1 105 [+ or -] 3.7 CSE 145 [+ or -] 3.2 **** 156 [+ or -] 5.1 **** CSE 132 [+ or -] 5.1 * 129 [+ or -] 5.1 CSE 128 [+ or -] 3.3 ** 124 [+ or -] 3.4 * SPE 149 [+ or -] 6.5 * 146 [+ or -] 5.7 **** SPE 141 [+ or -] 4.2 **** 138 [+ or -] 4 *** SPE 134 [+ or -] 1.7 **** 130 [+ or -] 2 **** * p < 0.05, ** p < 0.01, *** p < 0.005 and **** p < 0.001 in respect to the values "before injection" for each group.
Amadio P Jr, Cummings DM, Amadio P (1993) Nonsteroidal anti-inflammatory drugs. Tailoring therapy to achieve results and avoid toxicity. Postgrad Med 93: 73-76
Baytop T (1984) Therapy with medicinal plants in Turkey (Past and present). Publications of the Istanbul University, p 225, Istanbul
Bermejo-Benito P, Abad-Martinez MJ, Silvan-Sen AM, Sanz-Gomez A, Fernandez-Matellano L, Sanchez-Contreras S, Diaz-Lanza AM (1998) In vivo and in vitro antiinflammatory activity of saikosaponins. Life Sci 63: 1147-1156
Birch PJ, Harrison SM, Hayes AG, Rogers H, Tyers MB (1992) The non-peptide NK1 receptor antagonist, (+/-)-CP-96,345, produces antinociceptive and anti-oedema effects in the rat. Br J Pharmacol 105: 508-510
Campos MM, Mata LV, Calixto JB (1995) Expression of B1 kinin receptors mediating paw edema and formalin-induced nociception. Modulation by glucocorticoids. Can J Physiol Pharmacol 73: 812-819
Cioaca C, Margineanu C, Cucu V (1978) The saponins of Hedera helix with antibacterial activity. Pharmazie 33: 609-610
Cuellar MJ, Giner RM, Recio MC, Just MJ, Manez S, Cerda M, Hostettmann K, Rios JL (1997) Zanhasaponins A and B, antiphospholipase A2 saponins from an antiinflammatory extract of Zanha africana root bark. J Nat Prod 60: 1158-1160
DiRosa M, Giroud JP, Willoughby DA (1971) Studies of the mediators of the acute inflammatory response induced in rats in different sites by carrageenan and turpentine. J Pathol 104: 15-29
Dunne MW (1990) Pathophysiology: Concepts of Altered Health States with Contributors. Ed by Porth CM, Lippincott, Philadelphia, pp 165-176
Elias R, Diaz-Lanza AN, Vidal-Ollivier E, Balansard G, Faure R, Babadjamian A (1991) Triterpene saponins from leaves of Hedera helix. J Natl Prod 54: 98-103
Fung HB, Kirschenbaum HL (1999) Selective cyclooxygenase-2 inhibitors for the treatment of arthritis. Clin Ther 21: 1131-1157
Gamache DA, Povlishock JT, Ellis EF (1986) Carrageenan-induced brain inflammation. Characterization of the model. J Neurosurg 65: 679-685
Garcia M, Fernandez E, Navarro JA, del Pozo MD, Fernandez de Correz L (1995) Allergic contact dermatitis from Hedera helix L. Contact Dermatitis 33: 133-134
Garcia-Pastor P, Randazzo A, Gomez-Paloma L, Alcaraz MJ, Paya, M (1999) Effects of petrosaspongiolide M, a novel phospholipase A2 inhibitor, on acute and chronic inflammation. J Pharmacol Exp Ther 289: 166-172
Gilman SC, Carlson RP, Chang J, Lewis AJ (1985) The antiinflammatory activity of the immunomodulator Wy-18,251 (3-(p-chlorophenyl)thiazolo[3,2-a]benzimidazole2-acetic acid). Agents Actions 17: 53-59
Hausen BM, Brohan J, Konig WA, Faasch H, Hahn H, Bruhn, G (1987) Allergic and irritant contact dermatitis from falcarinol and didehydrofalcarinol in common ivy (Hedera helix L.). Contact Dermatitis 17: 1-9
Insel PA (1996) Analgesic, Antipyretic and Antiinflammatory Agents and Drugs Employed in the Treatment of Gout. ed. By Hardman JG, Limbird LE, McGraw-Hill, New York, pp 617-657
Ionac M, Parnham MJ, Plauchithiu M, Brune K (1996) Oxaceprol, an atypical inhibitor of inflammation and joint damage. Pharmacol Res 33: 367-373
Julien J, Gasquet M, Maillard C, Balansard G, Timon-David P (1985) Extracts of the ivy plant, Hedera helix, and their anthelminthic activity on liver flukes. Planta Med 17: 205-208
Kulkarni SK, Mehta AK, Kunchandy J (1986) Anti-inflammatory actions of clonidine, guanfacine and B-HT 920 against various inflammagen-induced acute paw oedema in rats. Arch Int Pharmacodyn Ther 279: 324-334
Leme JG, Hamamura L, Leite MP, Silva MR (1973) Pharmacological analysis of the acute inflammatory process induced in the rats paw by local injection of carrageenan and by heating. Br J Pharmacol 48: 88-96
Majester-Savornin B, Elias R, Diaz-Lanza AM, Balansard G, Gasquet M, Delmas F (1991) Saponins of the ivy plant, Hedera helix, and their leishmanicidic activity. Planta Med 57: 260-262
Massmanian A, Valcuende-Cavero F, Ramirez-Bosca A, Castells-Rodellas A (1988) Contact dermatitis from variegated ivy (Hedera helix subsp, canariensis Willd.). Contact Dermatitis 18: 247-248
Moulin-Traffort J, Favel A, Elias R, Regli P (1998) Study of the action of alpha-hederin on the ultrastructure of Candida albicans. Mycoses 41: 411-416
Suleyman H, Demirezer LO, Kuruuzum A, Banoglu ZN, Gocer F, Ozbakir G, Gepdiremen A (1999) Antiinflammatory effect of the aqueous extract from Rumex patientia L. roots. J Ethnopharmacol 65: 141-148
Szolcsanyi J, Helyes Z, Oroszi G, Nemeth J, Pinter E (1998) Release of somatostatin and its role in the mediation of the anti-inflammatory effect induced by antidromic stimulation of sensory fibres of rat sciatic nerve. Br J Pharmacol 123: 936-942
Trute A, Gross J, Mutschler E, Nahrstedt A (1997) In vitro antispasmodic compounds of the dry extract obtained from Hedera helix. Planta Med 63: 125-129
H. Suleyman, Ataturk University, Medical Faculty, Department of Pharmacology, TR-25240 Erzurum, Turkey Tel.: ++90-442-2361212/2423; Fax: ++90-442-2360968; e-mail: email@example.com
H. Suleyman (1), V. Mshvildadze (2), A. Gepdiremen (1), and R. Elias (3)
(1) Ataturk University, Medical Faculty, Department of Pharmacology, Erzurum, Turkey
(2) Institute of Pharmacochemistry, Tbilisi, Georgia
(3) Laboratory of Pharmacognosy and Homeopathy, Pharmacy Faculty of Mediterranean University, Marseille, 13385 Marseille, France
|Printer friendly Cite/link Email Feedback|
|Author:||Suleyman, H.; Mshvildadze, V.; Gepdiremen, A.; Elias, R.|
|Publication:||Phytomedicine: International Journal of Phytotherapy & Phytopharmacology|
|Date:||Jun 1, 2003|
|Previous Article:||A randomised double blind placebo controlled clinical trial of a standardised extract of fresh Crataegus berries (Crataegisan[R]) in the treatment of...|
|Next Article:||Anti-inflammatory activity of shikonin derivatives from Arnebia hispidissima.|