Hydroalcoholic extract of needles of Pinus eldarica enhances pentobarbital-induced sleep: possible involvement of GABAergic system

Document Type: Original Research Article


1 Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Pharmacological Research Center of Medicinal Plants, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

3 Neurocognitive Research Center, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran


Objective: Insomnia is accompanied by several health complications and the currently used soporific drugs can induce several side effects such as psychomotor impairment, amnesia, and tolerance. The present study was planned to investigate the sleep prolonging effect of Pinus eldarica.
Materials and Methods: Hydroalcoholic extract (HAE) of P. eldarica, its water fraction (WF), ethyl acetate fraction (EAF) and n-butanol fraction (NBF) were injected (intraperitoneally) to mice 30 min before administration of pentobarbital. Then, the latent period and continuous sleeping time were recorded. Also, LD50 of P. eldarica extract was determined and the possible neurotoxicity of the extract was tested on neural PC12 cells.
Results: The HAE and NBF decreased the latency of sleep (p<0.05) and significantly increased duration of sleep (p<0.05) induced by pentobarbital. These effects of P. eldarica were reversed by flumazenil. The LD50 value for HAE was found to be 4.8 g/Kg. HAE and its fractions did not show neurotoxic effects in cultured PC12-cell line.
Conclusion: The present data indicate that P. eldarica potentiated pentobarbital hypnosis without major toxic effect. Most probably, the main components responsible for this effect are non-polar agents which are found in NBF of this plant.


Main Subjects

Akhila JS, Shyamjith D, Alwar MC. 2007. Acute toxicity studies and determination of median lethal dose. Curr Sci, 93: 917-920.

Currie G, Wheat J. 2007. Insomnia: An overview of herbal treatments. Internet J Altern Med, 5.

Datta S.2010. Cellular and chemical neuroscience of mammalian sleep. Sleep Med, 11: 431-440.

Edewor-Kuponiyi TI. 2013. Plant-derived compounds with potential sedative and anxiolytic activities. Int J Basic Appl Sci, 2: 63-78.

Emamghoreishi M, Heidari-Hamedani G. 2006. Sedative-hypnotic activity of extracts and essential oil of coriander seeds. Iran J Basic Med Sci, 31: 22-27.

Ghorbani A, Asadpour E, Sadeghnia HR. 2015. Mechanism of protective effect of lettuce against glucose/serum deprivation-induced neurotoxicity. Nutr Neurosci, 18: 103-109.

Ghorbani A, Hadjzadeh MR, Rajaei Z, Zendehbad SB. 2014.Effects of fenugreek seeds on adipogenesis and lipolysis in normal and diabetic rat.Pak J Biol Sci, 17: 523-528.

Ghorbani A, Youssofabad NJ, Rakhshandeh H. 2012. Effect of Viola tricolor on pentobarbital-induced sleep in mice. Afri J Pharm Pharmacol, 6: 2503-2509.

GhorbaniA, Rakhshandeh H, Sadeghnia HR. 2013. Potentiating effects of Lactuca sativa on pentobarbital-induced sleep. Iran J Pharm Res, 12: 401-406.

Gottesmann C. 2002. GABA mechanisms and sleep. Neuroscience, 111: 231-239.

Guri A, Kefalas P, Roussis V. 2006. Antioxidant potential of six pine species. Phytother Res, 20: 263-266.

Hosseini A, Ghorbani A, Sadeghnia HR, Rajabian A, Rakhshandeh H. 2014. Potentiating effects of Lactuca serriola on pentobarbital-induced sleep. Res Opin Anim Vet Sci, 4: 601-607.

Ishisaka A, Ichikawa S, Sakakibara H, Piskula MK, Nakamura T, Kato Y, Ito M, Miyamoto K, Tsuji A, Kawai Y, Terao J. 2011. Accumulation of orally administered quercetin in brain tissue and its antioxidative effects in rats. Free Radic Biol Med, 51: 1329-1336.

Kambe D, KotaniM, Yoshimoto M, Kaku S, Chaki S, Honda K. 2010. Effects of quercetin on the sleep–wake cycle in rats: Involvement of gamma-aminobutyric acid receptor type A in regulation of rapid eye movement sleep. Brain Res, 12: 83-88.

Karonen M, Hämäläinen M, Nieminen R, Klika KD, Loponen J, Ovcharenko VV, EevaMoilanen E, KaleviPihlaja K. 2004. Phenolic extractives from the bark of Pinus sylvestris L. and their effects on inflammatory mediators nitric oxide and prostaglandin E2. J Agric Food Chem, 52: 7532-7540.

Kaundun SS, Lebreton P, Fady B. 1997. Genetic differences between Pinus halepensis, Pinus brutiaa nd Pinus eldarica based on needle flavonoids. Biochem Syst Ecol, 25: 553-562.

Lee E.2003. Effect of powdered pine needle (Pinus densiflora Siebet Zucc.) on serum and liver lipid composition and antioxidative capacity in rats fed high oxidative fat. J Korean Soc Food Sci Nutr, 32: 926-930.

Li K, Li Q, Li J, Gao D, Zhang T, Han Z, Zheng F. 2007. Effect of procyanidins from Pinus koraiensis bark on growth inhibition and expression of PCNA and TNF-α in mice with U14 cervical cancer. Therapy,4: 685-690.

Mamedov N, Craker LE. 2001. Medicinal plants used for the treatment of bronchial asthma in Russia and Central Asia. J Herbs Spices Med Plants, 8: 91-117.

Mamedov N, Gardner Z, Craker LE. 2005. Medicinal plants used in Russia and Central Asia for the treatment of selected skin conditions. J Herbs Spices Med Plants, 11: 191-222.

Meckes M, Calzada F, Tortoriello J,  González JL, Martínez M. 1998. Terpenoids Isolated from Psidium guajava hexane extract with depressant activity on central nervous system. Phytother Res, 10: 600-603.

Meletis CD, Zabriskie N. 2008.Natural approaches for optimal sleep. J Altern Complement Med, 14: 181-188.

Michelozzi M, Tognetti R, Maggino F, Radicati M. 2008.Seasonal variations in monoterpene profiles and ecophysiological traits in Mediterranean pine species of group “halepensis”. the Italian Society of Silviculture and Forest Ecology, 1: 65-74.

Mirheydar H. 1993. Herbal Information: Usage of plants in prevention and treatment of diseases, Islamic Culture Press Center, pp. 40-40, Tehran, Iran.

Mortazavian SM, Ghorbani A, Hesari TG. 2012. Effect of hydro-alcoholic extract of Viola tricolor and its fractions on proliferation of uterine cervix carcinoma cells. Iran J Obst Gyncol Infertil, 15: 9-16.

Mortazavian SM, Ghorbani A. 2012. Antiproliferative effect of viola tricolor on neuroblastoma cells in vitro. Aust J Med Herbalism, 24: 93-96.

Murillo-Rodriguez E, Arias-Carrion O, Sanguino-Rodriguez K, Gonzalez-Arias M, Haro R. 2009. Mechanisms of sleep-wake cycle modulation. CNS Neurol Disord Drug Targets, 8: 245-253.

Paulke A, Schubert-Zsilavecz M, Wurglics M. 2006. Determination of St. John's wort flavonoid-metabolites in rat brain through high performance liquid chromatography coupled with fluorescence detection. J Chromatogr B Analyt Technol Biomed Life Sci, 832: 109-113.

Potta SP, Doss MX, Hescheler J, Sachinidis A. 2005. Epigallocatechin-3-gallate (EGCG): A structural target for the development of potential therapeutic drugs against anti-proliferative diseases. Drug Design Rev Online, 2: 85–91.

Pritchett K, Mulder GB. 2003. The rotarod. Contemp Top Lab Anim Sci, 42: 49.

Rakhshandah H, Shakeri MT, Ghasemzadeh MR. 2007. Comparative hypnotic effect of Rosa damascena fractions and diazepam in mice. Iran J Pharm Res, 6: 193-197.

Rakhshandeh H, Sadeghnia HR, Ghorbani A. 2012. Sleep-prolonging effect of Coriandrum sativum hydro-alcoholic extract in mice. Nat Prod Res, 26: 2095-2098.

Rohdewald P.A. 2002. review of the french maritime pine bark extract (Pycnogenol): A herbal medication with a diverse clinical pharmacology. Int J Clin Pharmacol Ther, 40: 158-168.

Roth T, Drake C. 2004. Evolution of insomnia: current statusand future direction. Sleep Med, 5: S23–S30.

Seidel V. 2006. Initial and Bulk Extraction. In: Sarker SD, Latif Z, Gray AI, editors. Natural Product Isolation, New Jersey: Humana Press. 27-46.

Shafiee-Nick R, Ghorbani A, Vafaee F, Rakhshandeh H. 2012. Chronic administration of a combination of six herbs inhibits the progression of hyperglycemia and decreases serum lipids and aspartate amino transferase activity in diabetic rats. Adv Pharmacol Sci, 2012: 2012.

Sharma M, Rauniar GP, Das BP. 2012. Experimental study of various central nervous system effects of eugenol in mice and rats. Health Renaissance, 10: 208- 214.

Tian S, Shi Y, Zhou X, Ge L, Upur H. 2011.Total polyphenolic (flavonoids) content and antioxidant capacity of different Ziziphora clinopodioides Lam. extracts. Pharmacogn Mag, 7: 65-68.

Vafaee F, Hosseini M, Sadeghinia HR, Hadjzadeh MA, Soukhtanloo M, Rahimi M. 2014. The effects of soy extract on spatial learning and memory damage induced by global ischemia in ovariectomised rats. Malays J Med Sci, 21: 19-30.

Vallières A, Ivers H, Bastien CH, Beaulieu-Bonneau S, Morin CM. Variability and predictability in sleep patterns ofchronic insomniacs. J Sleep Res, 14: 447- 453.

Virgili F, Kobuchi H, Packer L. 1998. Nitrogen monoxide (NO) metabolism: antioxidant properties and modulation of inducible NO synthase activity in activated macrophages by procyanidins extracted from Pinus maritime (Pycnogenol) In: Rice-Evans CA, editor. Flavonoids in Health and Disease. New York: Marcel Dekker Inc. 421-436.

Vuorela S, Kreander K, Karonen M, Nieminen R, Hämäläinen M, Galkin A, Laitinen L, Salminen JP, Moilanen E, Pihlaja K, Vuorela H, Vuorela P, Marina Heinonen M. 2005. Preclinical evaluation of rapeseed, raspberry, and pine bark phenolics for health related effects. J Agric Food Chem, 53: 5922-5931.

Wasowski C, Marder M. 2012. Flavonoids as GABAA receptor ligands: the whole story?.J Exp Pharm. 4: 9-24.

Youdim KA, Qaiser MZ, Begley DJ, Rice-Evans CA, Abbott NJ. 2004. Flavonoid permeability across an in situ model of the blood-brain barrier. Free Radic Biol Med, 36: 592-604.