Research in Pharmaceutical Sciences, December 2015; 10(6): 497-503 School of Pharmacy & Pharmaceutical Sciences Received: May 2014 Isfahan University of Medical Sciences Accepted: July 2014 Original Article Evaluation of anti-spasmodic effect of Peucedanum pastinacifolium extracts on rat’s ileum 1,* 2 1,2 Hassan Sadraei , Gholam Reza Asghari , and Monireh Motaqedi 1Department of Pharmacology and Toxicology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, I.R. Iran. 2Isfahan Pharmaceutical Sciences Research Center, Isfahan University of Medical Sciences, Isfahan, I.R .Iran. Abstract Genus Peucedanum belongs to the subfamily of Apioideae (Umbelliferae family) and is reported to have many medicinal properties. Several spiecis of Peucedum reported to have antispasmodic activity. Peucedanum pastinacifolium species grows in Iran. However, so far there is no report on its antispasmodic activity. The objective of this research was to investigate antispasmodic activities of P. pastinacifolium extract using in vitro isolated tissue techniques. Hydroalcoholic and hexanoic extracts were prepared by perculation method from aerial part of P. pastinacifolium. A portion of rat isolated ileum was suspended under 1g tension in Tyrode’s solution at 37 C and gassed with O2. Effects of extracts of P. pastinacifolium were studied on ileum contractions induced by KCl (80 mM), acetylcholine (ACh, 250 µM) and electrical field stimulation (EFS). The hydroalcoholic extract P. pastinacifolium concentration dependently inhibited the response to KCl (IC50=220 ± 30 g/ml), ACh (IC50=175 ± 15 g/ml) and EFS (IC50=95 ± 15 g/ml). The hexanoic extract of P. pastinacifolium also had inhibitory effect on ileum contraction induced by KCl (IC50=16 ± 2 g/ml), ACh (IC50=30 ± 5 g/ml) or EFS (IC50=11 ± 4 g/ml). From these experiments it was concluded that both hydroalcoholic and hexanoic extract of P. pastinacifolium contain substances which have antispasmodic activities but these substances are mainly concentrated in the hexanoic extract. Keywords: Peucedanum pastinacifolium; Ileum; Antispasmodic INTRODUCTION antihyperlipidemic and antihypertensive activities (4,5), In addition it is believed that Umbelliferae family is a large family of they also have antidiabetic, antimutogenic, flowering plants with more than 150 genus and antitumor and antioxidant properties (6-9). 3000 different species (1). Fruit, leaves, and Other pharmacological activities suggested for roots of some of this family plant such as this genus include antiplatelet, antimicrobial parsley, dill, cumin, and fennel are used in and antiallergic activities (10-12). Two species traditional folk medicine (2). Umbelliferae of P. Japonicum and P. praeruptorium are family consists of three main subfamilies reported to have antispasmodic activities (12). including Hydrocotgloideae, Saniculoideae, The antispasmodic activity is suggested to be and Apioideae (2). Genus Peucedanum due to the presence of essential oil and belongs to subfamily of Apioideae with 120 furanocoumarins (3,13-15). Additionally, other different species (2). Peucedanum genus is species of this genus have been used known as Razianeh Kohi in Persian and Hog’s traditionally in the treatment of cold and cough Fennel in English with 21 different species due to pathogenic wind-heat, accumulation of grows all over Iran (3). This genus are phlegm-heat in the lung; they have also been separated by Flora Iranica into five different used as anti-tussive, anti-asthma and a remedy species including Leutea, Demavendia, for angina (18). P. pastinacifolium Boiss. & Cervaria, Zeravschamia, and Jahreniopsis (3). Hausskn which is commonly known as “Alafe- Peucedanum genus is reported to have many Tofangchi” is popular in folk medicine for medicinal properties. For example they have having antilipidemic and antidiabetic effects *Corresponding author: H. Sadraei Tel: 0098 31 37927086, Fax: 0098 31 36680011 Email: [email protected] H. Sadraei et al. / RPS 2015; 10(6): 497-503 (4,16,17). It is an endemic species in central Pharmacy and Pharmaceutical Science at parts of Iran and grows on more or less rocky Isfahan University of Medical Sciences. Aerial and scree steep slopes. It is used by local part of the plant was dried in shade and cut inhabitants in central and western Iran as an into small pieces and then grounded to powder antihyperlipidemic vegetable which has been using electrical miller (Moulinex, France). The verified in vivo (4,13). In one study on this hydroalcoholic extract as polar fraction and plant it is proved that ethanolic extract of the hexanoic extract as non-polar fraction were aerial parts of P. pastinacifolium can decrease obtained by percolation (24) using ethanol total serum cholesterol, low-density (80%) and hexane (100%) respectively. The lipoprotein cholesterol, triglyceride levels, and solvent of the extracts was evaporated to atherogenic indices in hypercholesterolemic dryness and percentage yield of the dried rats after eight days (4). In another study, the extract was determined. ethanolic extract of the aerial parts of the same species was injected into streptozotocin- Antispasmodic studies induced diabetic rats and showed the In this research isolated tissue preparation amelioration of their blood lipid profile (13). A was used for the assessment of antispasmodic phytochemical investigation on acetone extract activity of P. pastinacifolium extracts. Ileum of P. pastinacifolium demonstrated three linear segment were prepared from male Wistar rats furanocoumarins, bergapten, isopimpinellin, (180–220 g), bred in School of Pharmacy and methoxsalen, and a phenylpropanoid animal house (Isfahan), in accordance with the named elemicin (15). Bergapten and internationally accepted principles for isopimpinellin have antifeedant and laboratory animal use and care, as antimicrobial effects (19,20). Also bergapten recommended by university authority is reported to have antimutagenic effects (21). (Committee for the Update of the Guide for Elemicin which can be found only in essential the Care and Use of Laboratory Animals, oil has different effects, among which, 2010) (25). antimicrobial and antifungal effects are the Longitudinal strips of ileum, 2-3 cm long, most prominent ones (22,23). were mounted under 1 g resting force in an The air-dried aerial parts of P. organ bath (Harvard, England) filled with pastinacifolium mainly provides about 0.2% of Tyrode's solution (see drug and solutions essential oil, whose main components are section) at 37 C and gassed with O2. The known to be elemicin (31.1%), limonene tissues were washed several times with fresh (11.6%), myristicin (8.2%), methyl eugenol Tyrode's solution and allowed to relax to a (6.1%), dillapiole (5.9%), trans-β-ocimene stable base line. Tissue contractions were (5.9%), and α-pinene (4.9%) (15). recorded on a Harvard Universal Oscillograph Although two Pecedanum species was (Harvard, England) pen recorder device. reported to have antispasmodic activities, Neuronal stimulation was induced using however, so far there is no report on electrical field stimulation technique (EFS, 6 antispasmodic activity of P. pastinacifilum. V and 50 Hz for 1s duration) as described Therefore, the objective of this research was to before (26,27). Contraction induced by single investigate antispasmodic activity of P. dose of acetylcholine (ACh, 250 µM, 30 s pastinacifolium extracts in vitro by contact time) was studied in a non-cumulative pharmacological technique. manner. After reproducible contractions were established, at 12 min intervals, successive METHOD AND MATERIALS concentrations of extracts was added directly into organ bath. In the case of KCl (80 mM), Plant materials and extracts preparation following stabilization of tonic contraction, P. P. pastinacifolium was collected from pastinacifolium extract was added Golpaygan mountain in flowering season in cumulatively into organ bath using two fold 2013. A voucher specimen (2831) was increments in concentration. Initially a number deposited in the herbarium of the School of of pilot experiments were carried out for 498 Antispasmodic effect of P. pastinacifolium extracts determination of effective concentration RESULTS ranges of extract of P. pastinacifolium. Then full concentration response curves were Pharmacognosy obtained for each drug using 8-10 different The yield of hydroalcoholic and hexanoic concentrations of the extracts. extracts of P. pastinacifolium were 19% and Following completion of the experiment, 0.8%, respectively. Both extracts had dark the tissues were washed with fresh Tyrode's greenish colour. solution and tested for reversibility of the response. All experiments were performed Pharmacology alongside of time-matched vehicle treated When ileum segment was suspended in the controls. organ bath it started to produce irregular spontaneous contractile activity. Following Drugs and solutions repeated washing, the ileum gradually relaxed Tyrode's solution composed of: NaCl, to a stable baseline over 10-20 min. The tissue 136.9; KCl, 2.68; CaCl2, 1.8; MgCl2, 1.05; contracted rapidly to EFS reaching a peak NaHCO3, 11.9; NaH2PO4, 0.42 and glucose, followed by partial relaxation which was then 5.55 (in mM) were made up in distilled water. followed by a second peak and then relaxed Unless stated, all chemicals and drugs were towards the baseline as described before from Merck, Germany. (28,29). Relaxant effect of P. pastinacifolium The following drugs were used in this extracts