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Hypericum triquetrifolium Turra. extract exhibits antiinflammatory activity in the rat
Journal of Ethnopharmacology 80 (2002) 207 /209 www.elsevier.com/locate/jethpharm
Hypericum triquetrifolium Turra. extract exhibits antiinflammatory activity in the rat Bintug Ozturk a, Sebnem Apaydin b,*, Esin Goldeli b, Iskender Ince b, Ulvi Zeybek a a
Department of Pharmaceutical Botany, Faculty of Pharmacy, Ege University, 35100 Bornova, Izmir, Turkey b Centre for Drug R&D and Pharmacokinetic Applications, Ege University, 35100 Bornova, Izmir, Turkey Accepted 8 February 2002
Abstract The aim of the present study was to explore the probable antiinflammatory effect of Hypericum triquetrifolium Turra. in a rat model of carrageenan induced inflammation. Male Wistar rats were treated intraperitoneally with 0.4% dimethylsulphoxide (DMSO) (as control group) and H. triquetrifolium extract (25, 50, 60 mg/kg), 30 min before 0.1 ml 1% carrageenan injection. Paw volume was measured before and 1, 2, 3, 4, 5 and 6 h after the injection of carrageenan. The results are expressed as the mean9/s.e. mean and the statistical significance of differences between groups was analyzed by One Way Analysis of Variance (ANOVA). The intraplantar injection of carrageenan caused a time-dependent paw edema in the rat although saline injection caused no swelling. Intraperitoneal administration of H. triquetrifolium extract (25, 50, 60 mg/kg) inhibited paw swelling dose-dependently at 2, 3, 4, 5 and 6 h after carrageenan injection (P B/0.05). We can conclude that H. triquetrifolium extract may exert an antiinflammatory effect in rats. # 2002 Elsevier Science Ireland Ltd. All rights reserved. Keywords: Hypericum triquetrifolium ; Antiinflammatory effect; Rat; Plant extract
1. Introduction Hypericum species are known by their antidepressant (Harrer and Schulz, 1994; Linde et al., 1996; Vorbach et al., 1997; Wheatly, 1997; Volz, 1997), analgesic (Apaydin et al., 1999), spasmolytic (Jakovljevic et al., 2000) antiviral and wound healing (Miller 1998) effects for many years. Extracts of Hypericum perforatum (St John’s wort) are licensed as antidepressants in different countries (Mu¨ller et al., 1997). On the other hand, antiinflammatory effects of Hypericum species are shown in only a limited number of studies (Shipochliev et al., 1981). In a previous study, it was mentioned that Hypericum triquetrifolium Turra. extract inhibited the second phase of formalin test in small doses such as 10 mg kg1 in mice. Since it was clearly shown that peripheral inflammatory processes were involved in the second phase of formalin test
* Corresponding author. Tel: 90-232-3392754; fax: 90-2323425088. E-mail address: [email protected] (S. Apaydin).
(Tjolsen et al., 1992) it can be suggested that the extract may exert antiinflammatory activity. The aim of the present study was to investigate the antiinflammatory activity of the total extract of H. triquetrifolium Turra.
2. Methodology Fresh plants from wild collections, gathered at the start of the flowering period in July 1999 from Karaali, Manisa (a city located in the western part of Turkey) were used. Mainly, the aerial parts of the plants that have a high proportion of buds and flowers were selected. The plant was identified by Professor Ozcan Secmen, Ph.D from Ege University, Faculty of Science, Department of Biology, Section of Botany. The voucher specimen of the plants used in the present study was kept for record in the herbarium of Ege University, Faculty of Pharmacy, Department of Pharmaceutical Botany (voucher no. 5435).
0378-8741/02/$ - see front matter # 2002 Elsevier Science Ireland Ltd. All rights reserved. PII: S 0 3 7 8 - 8 7 4 1 ( 0 2 ) 0 0 0 4 4 - 2
208
B. Ozturk et al. / Journal of Ethnopharmacology 80 (2002) 207 /209
2.1. Extraction of H. triquetrifolium Turra. The crude drug was dried in shade and fine powder of the plant was obtained by a mill (Brabender OHG, Duisburg). A modified method of Wagner and Bladt (1994) was used for the extraction of the powdered plant. Methanol at 80 8C was used for soxhlet extraction, using 750 ml methanol for 100 g crude drug and the extract was dried in vacuo (yield 38.22%) After the lyophilization of the extract (yield 78.07%), it was administered to rats immediately after dissolved in 10 ml of 0.4% DMSO. 2.2. Animals Male Wistar rats (200 /300 g) were used. Animals were housed in a room maintained at 229/1 8C with an alternating 12 h light-dark cycle. Food and water were available ad libitum. The animals were transported to a quiet laboratory at least 1 h before the experiment. 2.3. Assesment of antiinflammatory effect of H. triquetrifolium Turra. The antiinflammatory activity was evaluated by the carrageenan-induced paw edema test in the rat (Winter et al., 1962; Schapoval et al., 1998). Male Wistar rats were deprived of food overnight and treated intraperitoneally with 0.4% dimethylsulphoxide (DMSO) (as control group) and H. triquetrifolium extract (25, 50, 60 mg/kg), 30 min before 0.1 ml 1% carrageenan in isotonic saline was injected subplantarly into one of the hind paws. The contralateral paw was injected with 0.1 ml saline and used as a control. Paw volume was measured by water plethysmometer (Ugo Basile) before and 1, 2, 3, 4, 5 and 6 h after the injection of carrageenan into the plantar region of the right hind paw.
injection caused no swelling (data not shown). In carrageenan-induced paw edema in rats, intraperitoneal administration of H. triquetrifolium extract (25, 50, 60 mg/kg) inhibited paw swelling dose-dependently at 2, 3, 4, 5 and 6 h after carrageenan injection (P B/0.05). Percent increment in paw swelling was calculated by using the values before carrageenan injection (Fig. 1). Although Hypericum species are traditionally used to relieve inflammation (Miller, 1998), there are a few number of studies indicating the antiinflammatory effects of them. The effects of proinflammatory agents such as carrageenan and prostaglandin E1 were significantly inhibited by St John’s wort (H. perforatum L.) in Wistar albino rats (Shipochliev et al., 1981). The antiinflammatory activities of Hypericum lalandii (Recio et al., 1995) and a methanolic extract of Hypericum empetrifolium Willd. were demonstrated in rats (Trovato et al., 2001) by reducing the edema of carrageenan injection. Although we have demonstrated the antiinflammatory activity of H. triquetrifolium Turra. extract, the mechanism of this antiinflammatory effect could not be stated. The antiinflammatory effect of the plant extract may be due to its possible antioxidant activity since
2.4. Drugs The preparation of the plant extract was described above in detail. Carrageenan and DMSO were purchased from Sigma Chemical Co. (St. Louis, USA). 2.5. Statistical analysis The results are expressed as the mean9/s.e. mean and the statistical significance of differences between groups was analyzed by one way analysis of variance (ANOVA). P B/0.05 was considered as significant.
3. Results and discussion The intraplantar injection of carrageenan caused a time-dependent paw edema in the rat although saline
Fig. 1. Inhibition of carrageenan-induced paw edema by intraperitoneal administration of H. triquetrifolium Turra in rats. Data are expressed as mean and vertical lines show s.e. mean (n 10 for each group). * P B 0.05 versus control. Percent increment in paw swelling was calculated by using the values before carrageenan injection
B. Ozturk et al. / Journal of Ethnopharmacology 80 (2002) 207 /209
some of the Hypericum species such as Hypericum perforatum L. were clearly shown to exert strong antioxidant activity (Bolshakova et al., 1998; Tripathi and Pandey, 1999) by inhibiting free radical production (Hunt et al., 2001).
4. Conclusions The present data clearly showed that total extract of H. triquetrifolium Turra. exhibits antiinflammatory activity on inhibiting the edema formation after carrageenan subplantar injection in the rat. Further studies must be conducted in order to clarify the exact antiinflammatory mechanism of this plant extract and to figure out which constituent of the extract exerts this activity.
Acknowledgements This study is designed and performed in Ege University, Center for Drug R&D and Pharmacokinetic Applications.
References Apaydin, S., Zeybek, U., Ince, I., Elgin, G., Karamenderes, C., Ozturk, B., Tuglular, I. 1999. Hypericum triquetrifolium Turra extract exhibits antinociceptive activity in the mouse. Journal of Ethnopharmacology 67, 307 /312. Bolshakova, I.V., Lozovskaia, E.L., Sapezhinskii, I.I. 1998. Antioxidant properties of plant extracts. Biofizika 43, 186 /188. Harrer, G., Schulz, V. 1994. Clinical investigation of tha antidepressant effectiveness of Hypericum. Journal of Geriatric Psychiatry and Neurology 7 (Suppl 1), 6 /8. Hunt, E.J., Lester, C.E., Lester, E.A., Tackett, R.L. 2001. Effect of St John’s wort on free radical production. Life Sciences 69, 181 /190. Jakovljevic, V., Popovic, M., Mimica-Dukic, N., Sabo, A., Gvozdenovic, I. 2000. Pharmacodynamic study of Hypericum perforatum L. Phytomedicine 7, 449 /453.
209
Linde, K., Ramirez, G., Mulrow, C.D., Pauls, A., Weidenhammer, W., Melchart, D. 1996. St John’s wort for depression */an overview and meta-analysis of randomized clinical trials. British Medical Journal 313, 253 /258. Miller, A.L. 1998. St John’s Wort (Hypericum perforatum ): clinical effects on depression and other conditions. Alternative Medicine Reviews 3, 18 /26. Mu¨ller, W.E., Rolli, M., Schafer, C., Hafner, U. 1997. Effects of hypericum extract (LI 160) in biochemical models of antidepressant activity. Pharmacopsychiatry 30 (Suppl 2), 102 /107. Recio, M.C., Giner, R.M., Manez, S., Rios, J.L., Marston, A., Hostettmann, K. 1995. Screening of tropical medicinal plants for antiinflammatory activity. Phytotherapy Research 9, 571 /574. Schapoval, E., de Vargas, M., Chaves, C., Bridi, R., Zuanazzi, J., Henriques, A. 1998. Antiinflammatory and antinociceptive activities of extracts and isolated compounds from Stachytarpheta cayennensis . Journal of Ethnopharmacology 60, 53 /59. Shipochliev, T., Dimitrov, A., Aleksandrova, E. 1981. Antiinflammatory action of a group of plant extracts. Veterinary Medicine Nauki 18, 87 /94. Tjolsen, A., Berge, O., Hunskaar, S., Rosland, J.H., Hole, K. 1992. The formalin test: an evaluation of the method. Pain 51, 5 /17. Trovato, A., Raneri, E., Kouladis, M., Tzakou, O., Taviano, M.F., Galati, E.M. 2001. Anti inflammatory and analgesic activity of Hypericum empetrifolium Willd. (Guttiferae). Farmaco 56, 455 / 457. Tripathi, Y.B., Pandey, E. 1999. Role of alcoholic extract of shoot of Hypericum perforatum on lipid peroxidation and various species of free radicals in rats. Indian Journal of Experimental Biology 37, 567 /571. Volz, H.P. 1997. Controlled clinical trials of Hypericum extracts in depressed patients */an overview. Pharmacopsychiatry 30 (Suppl 2), 72 /76. Vorbach, E.U., Arnoldt, K.H., Hubner, W.D. 1997. Efficacy and tolerability of St John’s wort extract LI 160 versus imipramine in patients with severe depressive episodes according to ICD-10. Pharmacopsychiatry 30 (Suppl 2), 81 /85. Wagner, I., Bladt, S. 1994. Pharmacetical quality of hypericum extracts. Journal of Geriatric Psychiatry and Neurology. 7 (Suppl 1), 65 /68. Wheatly, D. 1997. LI 160, an extract of St John’s wort, versus amitriptyline in mildly to moderately depressed outpatients */a controlled 6-week clinical trial. Pharmacopsychiatry 30 (Suppl 2), 77 /80. Winter, C.A., Risley, E.A., Nuss, G.W. 1962. Carrageenan-induced edema in hind paw of rat as an assay for antiinflammatory drugs. Progress Society Biological Medicine 11, 544 /547.
Hypericum triquetrifolium Turra. extract exhibits antiinflammatory activity in the rat Bintug Ozturk a, Sebnem Apaydin b,*, Esin Goldeli b, Iskender Ince b, Ulvi Zeybek a a
Department of Pharmaceutical Botany, Faculty of Pharmacy, Ege University, 35100 Bornova, Izmir, Turkey b Centre for Drug R&D and Pharmacokinetic Applications, Ege University, 35100 Bornova, Izmir, Turkey Accepted 8 February 2002
Abstract The aim of the present study was to explore the probable antiinflammatory effect of Hypericum triquetrifolium Turra. in a rat model of carrageenan induced inflammation. Male Wistar rats were treated intraperitoneally with 0.4% dimethylsulphoxide (DMSO) (as control group) and H. triquetrifolium extract (25, 50, 60 mg/kg), 30 min before 0.1 ml 1% carrageenan injection. Paw volume was measured before and 1, 2, 3, 4, 5 and 6 h after the injection of carrageenan. The results are expressed as the mean9/s.e. mean and the statistical significance of differences between groups was analyzed by One Way Analysis of Variance (ANOVA). The intraplantar injection of carrageenan caused a time-dependent paw edema in the rat although saline injection caused no swelling. Intraperitoneal administration of H. triquetrifolium extract (25, 50, 60 mg/kg) inhibited paw swelling dose-dependently at 2, 3, 4, 5 and 6 h after carrageenan injection (P B/0.05). We can conclude that H. triquetrifolium extract may exert an antiinflammatory effect in rats. # 2002 Elsevier Science Ireland Ltd. All rights reserved. Keywords: Hypericum triquetrifolium ; Antiinflammatory effect; Rat; Plant extract
1. Introduction Hypericum species are known by their antidepressant (Harrer and Schulz, 1994; Linde et al., 1996; Vorbach et al., 1997; Wheatly, 1997; Volz, 1997), analgesic (Apaydin et al., 1999), spasmolytic (Jakovljevic et al., 2000) antiviral and wound healing (Miller 1998) effects for many years. Extracts of Hypericum perforatum (St John’s wort) are licensed as antidepressants in different countries (Mu¨ller et al., 1997). On the other hand, antiinflammatory effects of Hypericum species are shown in only a limited number of studies (Shipochliev et al., 1981). In a previous study, it was mentioned that Hypericum triquetrifolium Turra. extract inhibited the second phase of formalin test in small doses such as 10 mg kg1 in mice. Since it was clearly shown that peripheral inflammatory processes were involved in the second phase of formalin test
* Corresponding author. Tel: 90-232-3392754; fax: 90-2323425088. E-mail address: [email protected] (S. Apaydin).
(Tjolsen et al., 1992) it can be suggested that the extract may exert antiinflammatory activity. The aim of the present study was to investigate the antiinflammatory activity of the total extract of H. triquetrifolium Turra.
2. Methodology Fresh plants from wild collections, gathered at the start of the flowering period in July 1999 from Karaali, Manisa (a city located in the western part of Turkey) were used. Mainly, the aerial parts of the plants that have a high proportion of buds and flowers were selected. The plant was identified by Professor Ozcan Secmen, Ph.D from Ege University, Faculty of Science, Department of Biology, Section of Botany. The voucher specimen of the plants used in the present study was kept for record in the herbarium of Ege University, Faculty of Pharmacy, Department of Pharmaceutical Botany (voucher no. 5435).
0378-8741/02/$ - see front matter # 2002 Elsevier Science Ireland Ltd. All rights reserved. PII: S 0 3 7 8 - 8 7 4 1 ( 0 2 ) 0 0 0 4 4 - 2
208
B. Ozturk et al. / Journal of Ethnopharmacology 80 (2002) 207 /209
2.1. Extraction of H. triquetrifolium Turra. The crude drug was dried in shade and fine powder of the plant was obtained by a mill (Brabender OHG, Duisburg). A modified method of Wagner and Bladt (1994) was used for the extraction of the powdered plant. Methanol at 80 8C was used for soxhlet extraction, using 750 ml methanol for 100 g crude drug and the extract was dried in vacuo (yield 38.22%) After the lyophilization of the extract (yield 78.07%), it was administered to rats immediately after dissolved in 10 ml of 0.4% DMSO. 2.2. Animals Male Wistar rats (200 /300 g) were used. Animals were housed in a room maintained at 229/1 8C with an alternating 12 h light-dark cycle. Food and water were available ad libitum. The animals were transported to a quiet laboratory at least 1 h before the experiment. 2.3. Assesment of antiinflammatory effect of H. triquetrifolium Turra. The antiinflammatory activity was evaluated by the carrageenan-induced paw edema test in the rat (Winter et al., 1962; Schapoval et al., 1998). Male Wistar rats were deprived of food overnight and treated intraperitoneally with 0.4% dimethylsulphoxide (DMSO) (as control group) and H. triquetrifolium extract (25, 50, 60 mg/kg), 30 min before 0.1 ml 1% carrageenan in isotonic saline was injected subplantarly into one of the hind paws. The contralateral paw was injected with 0.1 ml saline and used as a control. Paw volume was measured by water plethysmometer (Ugo Basile) before and 1, 2, 3, 4, 5 and 6 h after the injection of carrageenan into the plantar region of the right hind paw.
injection caused no swelling (data not shown). In carrageenan-induced paw edema in rats, intraperitoneal administration of H. triquetrifolium extract (25, 50, 60 mg/kg) inhibited paw swelling dose-dependently at 2, 3, 4, 5 and 6 h after carrageenan injection (P B/0.05). Percent increment in paw swelling was calculated by using the values before carrageenan injection (Fig. 1). Although Hypericum species are traditionally used to relieve inflammation (Miller, 1998), there are a few number of studies indicating the antiinflammatory effects of them. The effects of proinflammatory agents such as carrageenan and prostaglandin E1 were significantly inhibited by St John’s wort (H. perforatum L.) in Wistar albino rats (Shipochliev et al., 1981). The antiinflammatory activities of Hypericum lalandii (Recio et al., 1995) and a methanolic extract of Hypericum empetrifolium Willd. were demonstrated in rats (Trovato et al., 2001) by reducing the edema of carrageenan injection. Although we have demonstrated the antiinflammatory activity of H. triquetrifolium Turra. extract, the mechanism of this antiinflammatory effect could not be stated. The antiinflammatory effect of the plant extract may be due to its possible antioxidant activity since
2.4. Drugs The preparation of the plant extract was described above in detail. Carrageenan and DMSO were purchased from Sigma Chemical Co. (St. Louis, USA). 2.5. Statistical analysis The results are expressed as the mean9/s.e. mean and the statistical significance of differences between groups was analyzed by one way analysis of variance (ANOVA). P B/0.05 was considered as significant.
3. Results and discussion The intraplantar injection of carrageenan caused a time-dependent paw edema in the rat although saline
Fig. 1. Inhibition of carrageenan-induced paw edema by intraperitoneal administration of H. triquetrifolium Turra in rats. Data are expressed as mean and vertical lines show s.e. mean (n 10 for each group). * P B 0.05 versus control. Percent increment in paw swelling was calculated by using the values before carrageenan injection
B. Ozturk et al. / Journal of Ethnopharmacology 80 (2002) 207 /209
some of the Hypericum species such as Hypericum perforatum L. were clearly shown to exert strong antioxidant activity (Bolshakova et al., 1998; Tripathi and Pandey, 1999) by inhibiting free radical production (Hunt et al., 2001).
4. Conclusions The present data clearly showed that total extract of H. triquetrifolium Turra. exhibits antiinflammatory activity on inhibiting the edema formation after carrageenan subplantar injection in the rat. Further studies must be conducted in order to clarify the exact antiinflammatory mechanism of this plant extract and to figure out which constituent of the extract exerts this activity.
Acknowledgements This study is designed and performed in Ege University, Center for Drug R&D and Pharmacokinetic Applications.
References Apaydin, S., Zeybek, U., Ince, I., Elgin, G., Karamenderes, C., Ozturk, B., Tuglular, I. 1999. Hypericum triquetrifolium Turra extract exhibits antinociceptive activity in the mouse. Journal of Ethnopharmacology 67, 307 /312. Bolshakova, I.V., Lozovskaia, E.L., Sapezhinskii, I.I. 1998. Antioxidant properties of plant extracts. Biofizika 43, 186 /188. Harrer, G., Schulz, V. 1994. Clinical investigation of tha antidepressant effectiveness of Hypericum. Journal of Geriatric Psychiatry and Neurology 7 (Suppl 1), 6 /8. Hunt, E.J., Lester, C.E., Lester, E.A., Tackett, R.L. 2001. Effect of St John’s wort on free radical production. Life Sciences 69, 181 /190. Jakovljevic, V., Popovic, M., Mimica-Dukic, N., Sabo, A., Gvozdenovic, I. 2000. Pharmacodynamic study of Hypericum perforatum L. Phytomedicine 7, 449 /453.
209
Linde, K., Ramirez, G., Mulrow, C.D., Pauls, A., Weidenhammer, W., Melchart, D. 1996. St John’s wort for depression */an overview and meta-analysis of randomized clinical trials. British Medical Journal 313, 253 /258. Miller, A.L. 1998. St John’s Wort (Hypericum perforatum ): clinical effects on depression and other conditions. Alternative Medicine Reviews 3, 18 /26. Mu¨ller, W.E., Rolli, M., Schafer, C., Hafner, U. 1997. Effects of hypericum extract (LI 160) in biochemical models of antidepressant activity. Pharmacopsychiatry 30 (Suppl 2), 102 /107. Recio, M.C., Giner, R.M., Manez, S., Rios, J.L., Marston, A., Hostettmann, K. 1995. Screening of tropical medicinal plants for antiinflammatory activity. Phytotherapy Research 9, 571 /574. Schapoval, E., de Vargas, M., Chaves, C., Bridi, R., Zuanazzi, J., Henriques, A. 1998. Antiinflammatory and antinociceptive activities of extracts and isolated compounds from Stachytarpheta cayennensis . Journal of Ethnopharmacology 60, 53 /59. Shipochliev, T., Dimitrov, A., Aleksandrova, E. 1981. Antiinflammatory action of a group of plant extracts. Veterinary Medicine Nauki 18, 87 /94. Tjolsen, A., Berge, O., Hunskaar, S., Rosland, J.H., Hole, K. 1992. The formalin test: an evaluation of the method. Pain 51, 5 /17. Trovato, A., Raneri, E., Kouladis, M., Tzakou, O., Taviano, M.F., Galati, E.M. 2001. Anti inflammatory and analgesic activity of Hypericum empetrifolium Willd. (Guttiferae). Farmaco 56, 455 / 457. Tripathi, Y.B., Pandey, E. 1999. Role of alcoholic extract of shoot of Hypericum perforatum on lipid peroxidation and various species of free radicals in rats. Indian Journal of Experimental Biology 37, 567 /571. Volz, H.P. 1997. Controlled clinical trials of Hypericum extracts in depressed patients */an overview. Pharmacopsychiatry 30 (Suppl 2), 72 /76. Vorbach, E.U., Arnoldt, K.H., Hubner, W.D. 1997. Efficacy and tolerability of St John’s wort extract LI 160 versus imipramine in patients with severe depressive episodes according to ICD-10. Pharmacopsychiatry 30 (Suppl 2), 81 /85. Wagner, I., Bladt, S. 1994. Pharmacetical quality of hypericum extracts. Journal of Geriatric Psychiatry and Neurology. 7 (Suppl 1), 65 /68. Wheatly, D. 1997. LI 160, an extract of St John’s wort, versus amitriptyline in mildly to moderately depressed outpatients */a controlled 6-week clinical trial. Pharmacopsychiatry 30 (Suppl 2), 77 /80. Winter, C.A., Risley, E.A., Nuss, G.W. 1962. Carrageenan-induced edema in hind paw of rat as an assay for antiinflammatory drugs. Progress Society Biological Medicine 11, 544 /547.