Abstract
The effects of different extracts of parsley and celery leaves (Et2O, CHCl3, EtOAC, n.-BuOH, and H2O) on ketamine-induced sleeping time have been investigated. The experiments were conducted on BALB/C white laboratory mice divided in five groups. On the first day, each group received ketamine 40 mg/ml s.c. After 5 t1/2 (next day), mice were treated with 10% extracts (Et2O, CHCl3, EtOAC, n.-BuOH, H2O) of parsley and celery leaves, 1 ml/kg i.p. in two doses, each after 2-h interval. One hour after the last dose, animals received ketamine 40 mg/kg s.c. Just after administration of ketamine, induction time and time of sleeping were measured. Most of the examined extracts of both plants exhibited insignificant changes in induction time. Only the EtOAc extract of celery exhibited significant increase, whereas Et2O and n.-BuOH extract of parsley induced significant decrease of induction time in treated animals. Furthermore, all examined extracts of both plants exhibited increase of sleeping time in animals treated with ketamine. The Et2O extract of both celery and parsley exhibited the highest effect. Generally, examined celery extracts caused prolongation of sleeping time in animals compared with those obtained for parsley.
Introduction
Parsley [Petroselinum crispum. (Mill.) Nym. ex A.W.Hill] and celery (Apium graveolens. L.) are well-known spicy aromatic plants belonging to the Apiaceae family. Both of these plants are widely used as a fragrance in different food products. They are also used in traditional and contemporary medicine in treatment of different ailments and complaints. Healing properties and medical use are mostly related to a wide range of active biomolecules present in plant, primarily flavonoids and essential oil.
Major flavonoids in both herbs are apiin and luteolin. However, presence of significant amount of apigenin in parsley is also confirmed (Nielsen et al., Citation1999; Fejes et al., Citation2000). Major compounds of parsley essential oil are apiol, myristicin, limonene, and 1,3,8-p.-menthatriene; other components are α.- and β.-pinene, camphene, terpinolene, β.-phelandrene, and myrcene (Bruneton, Citation1999). Essential oil of celery also contains myristicin, especially obtained from roots. Other compounds of essential oil are limonene, α.- and β.-selinene, santalol, and α.- and β.-eudesmol. Parsley contains a wide range of furanocoumarins (psoralen, bergapten, isoimperatorin, oxypeucedanin, xanthoxin, trioxalen, and angelicin) (Wagner & Bladt, Citation1996; Lombaert et al., Citation2001), polienes (farcarinol), proteins, and sugars. Also, parsley contains a very high level of vitamins A and C, some of vitamins of B complex, calcium, and iron. A wide range of furanocoumarins, such as bergapten, psoralen, xanthoxin, isoimperatorin, isopimpinellin, apiumosid, and celerosid, is also found in celery herb (Wagner & Bladt, Citation1996). In celery, high levels of vitamins, minerals, and fatty acids are also present (Leung & Foster, Citation1996).
In medical treatment, parsley is used as a carminative, spasmolytic, emenagogue, and antireumatic, for treatment of dyspepsia, cystitis, dysmenorrhea, functional amenorrhea, and myalgia (Wichtl & Bisset, Citation1994; Blumenthal, Citation1999). Different compounds present in celery root exhibit anti-inflammatory and analgesic activity (Lewis et al., Citation1985; Atta et al., Citation1998). In vivo. examinations indicate hepatoprotective effects of celery in paracetamol and thioacetamide intoxication (Singh & Handa, Citation1995). Protective effects of celery are also confirmed in cellular ishemia (Zhang et al., Citation1999). Some investigations are also related to the antioxidant properties of apigenin (Fejes et al., Citation1998 Citation2000). Apigenin expressed notable antiplatelet activities (Teng et al., Citation1985). For methoxalene, a compound present in both plants, the effect on hypnotic and subhypnotic doses of pentobarbital in mice is confirmed (Jakovljevic et al., Citation2002).
The well-known toxicity of parsley and celery and their essential oils is still not completely clarified. Earlier, it was believed that apiol and myristicin are the compounds responsible for toxicity. However, recent data indicate potential anticancer activity of myristicin (Steimetz & Potter, Citation1991; Zheng et al., Citation1993). This compound induces increased activity of glutathione-S.-transferase, which catalyzes the glutathione reaction with electrophyles, activated by carcinogens, thus yielding less toxic conjugates that are immediately eliminated via excretion.
Studies on effects on sleeping time of parsley and celery have not been carried out so far. In this investigation, activity of different extracts of parsley and celery leaves on induction and sleeping time induced by ketamine (40 mg/kg s.c.) in mice are reported.
Materials and Methods
Plant material
Parsley [Petroselinum crispum. (Mill.) Nym. ex A.W. Hill, Apiaceae] and celery (Apium graveolens. L.; Apiaceae), were collected in June 2003 in the Vojvodina province of Serbia and Montenegro. Voucher specimens (Ph-A no. 22/03 and Ph-A no. 12/03) were confirmed and deposited at the Herbarium of the Department of Pharmacognosy, Faculty of Medicine, University of Novi Sad.
Preparation of extracts
Extracts of both parsley and celery were extracted from 50 g of fresh leaves. Plant material was macerated three times in 70% methanol (MeOH) during a 24-h period. The macerates were collected, filtered, and evaporated to dryness under vacuum. The residues were dissolved in water and successively extracted with four solvents of increasing polarity: ether (Et2O), chloroform (CHCl3), ethylacetate (EtOAc), and n.-butanol (n.-BuOH). The extraction was carried out until a colorless extract was obtained. The residue was the aqueous extract. All of five extracts (Et2O, CHCl3, EtOAc, n.-BuOH, and H2O) were evaporated to dryness and then dissolved in 50% ethanol to make 10% (w/v) solutions. Both initial extracts and their diluted solutions were further used for research.
Effects on sleeping time of prepared parsley and celery extracts was studied on white laboratory mice (type BALB/C, body weight 20–28 g) of both genders. Mice were divided into five groups, consisting of two subgroups, and containing five animals each. Animals were kept under constant environmental conditions (room temperature 21 ± 1°C; humidity 55 ± 1.5%, with 12-h light period) in our own breeding stock.
During the first day, animals of each group received ketamine 40 mg/ml s.c. (Ralatek). Induction time and sleeping time were measured after application of the drug. After 5 t1/2 (next day), mice were treated with obtained 10% (Et2O, CHCl3, EtOAC, n.-BuOH, H2O) extracts of parsley or celery, 1 ml/kg i.p., in two doses. Each dose was administered after a 2-h interval. One hour after receipt of the last dose of extract, animals were treated with ketamine, 40 mg/kg s.c. Induction time and time of sleeping were measured just after the administration of ketamine.
The results are presented as mean values ± standard deviation (SD). The significant differences were analyzed by Student's t.-test.
Results and Discussion
The following figures present the results of the induction time and the time of sleeping after treatment of mice with ketamine 40 mg/ml s.c., together with the extracts of parsley and celery leaves. Animals received 10% solutions of Et2O, CHCl3, EtOAc, n.-BuOH, or H2O extracts in dose of 1 ml/kg (i.p.).
In , effects of celery extracts (Et2O, CHCl3, EtOAc, n.-BuOH, or H2O) on induction time are presented. As can be seen, Et2O, CHCl3, n.-BuOH, and water extract have not exhibited notable changes in induction time of mice after receiving ketamine. Only the EtOAc extract exhibited significant increase of induction time in animals.
Figure 1 Effect of celery leaves extract on induction time in mice.
Effects of celery extracts on the sleeping time are presented in . All examined extracts showed very significant influence on the sleeping time of treated animals. However, the highest effects were exhibited by Et2O and n.-BuOH extracts.
Figure 2 Effect of celery leaves extract on sleeping time in mice.
shows the activities of examined parsley leaves extracts (Et2O, CHCl3, EtOAc, n.-BuOH, or H2O) on induction time in mice. Three examined extracts (CHCl3, EtOAc, and H2O) exhibited no notable effect on induction time after treatment of animals with ketamine. Only the Et2O and n.-BuOH extract expressed significant decrease of induction time.
Figure 3 Effect of parsley leaves extract on induction time in mice.
The obtained results indicate certain similarities in activity of parsley () and celery leaves extracts () on the induction time. Most of examined extracts exhibited insignificant changes in induction time. However, only EtOAc extract of celery exhibited significant increase of induction time. On the contrary, Et2O and n.-BuOH extract of parsley expressed significant decrease of induction time in treated animals.
Effects of parsley extracts on the time of sleeping are presented in . All examined extracts showed increase of sleeping time in animals treated with ketamine. This activity is very similar to those obtained for celery leaves extracts. However, the highest effect was expressed by Et2O extract (more than 300%).
Figure 4 Effect of parsley leaves extract on sleeping time in mice.
Comparing the results obtained in examination of effects of celery () and parsley extracts () on sleeping time, it is obvious that both plants exhibited similar activities. The Et2O extract of both plants exhibited the highest effect. However, examined celery extracts generally exhibited stronger activities compared with those obtained by parsley.
From the obtained results, it can be concluded that most of the examined celery and parsley extracts exhibited no significant effects on the induction time in mice trated with ketamine (40 mg/kg s.c.). On the other hand, both plants caused prolongation of sleeping time in animals.
Acknowledgments
This research was supported by the Ministry of Sciences, Technology and Development, Republic of Serbia, Project no. 1862.
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