Pumpkin

Cucurbita pepo L. (pumpkin) belongs to the melon family Cucurbitaceae which comprises approximately 95 genera and 950-980 species (Schaefer and Renner 2011). Cucurbita pepo is indigenous to warm and temperate regions of Central and North America and is cultivated there. It also exists in wild form in Europe and Asia. The origin is uncertain. The common ancestor of all the current Cucurbita pepo varieties originates probably from Mexico as confirmed by archeological findings (Andres 2003). The herbal substance

(whole, dried, ripe seed) is mentioned in several well –known handbooks, such as Madaus (1938), Martindale (2007), Wichtl (2004), Gruenwald et al. (2000), Duke’s Handbook of Medicinal Herbs (Duke 2000), WHO Monographs (2009) and ESCOP Monographs (2009).  The seeds and oil from pumpkin seeds have been used for many years for the relief of difficulties associated with an enlarged prostate gland and micturition problems related to overactive bladder. The pumpkin seeds yield approximately 50% fatty oil, (mostly linoleic and oleic acid and tocopherol), but the putative active constituents are ∆7-sterols (avenasterol, spinasterol) and ∆5-sterols (sitosterol, stigmasterol).

Constituents of the pumpkin seeds

Fatty oil

The fatty oil is obtained from comminuted seeds which are roasted immediately before pressing.The physical -chemical characteristics of the oil and its content of fatty acids, tocopherols, carotenoids, chlorophyll pigments , squalene and sterols are described in the literature (Bombardelli and Morazzoni 1997; Fruhwirth and Hermetter 2008; Fruhwirth et al. 2003). The fatty oil content of pumpkin seed is about 50% (45-60%) (Murkovic et al. 1996; Tsaknis et al. 1997; Pumpkin (Cucurbita pepo L.) seed oil is dark green and has a high content of free fatty acids (Murkovic et al. 1996; Tsaknis et al. 1997). Due to its colour and the foam formation, the oil is not used for cooking (Murkovic et al. 2004). When obtained by pressing, the oil has a dark red to green colour (due to content of carotenoids and chlorophylls), a red fluorescence and a nutty taste

Content of unsaponifiable fraction

Sterols, up to 0.5% of the oil (55-60% of the unsaponifiable fraction), predominantly ∆7sterols (∆7 or delta -7, signifies a double bond between C-7 and C-8). Much smaller amounts of ∆5- and ∆8 -sterols are also present (Bastic et al. 1977).

Four ∆7-sterols account for 75-88% of the total sterols (Tsaknis et al. 1997; Bastic et al.

1977; Akihisa et al. 1986) and 40-50% of the unsaponifiable part of the oil:

∆7,25- stigmastadienol (stigmasta -7,25-dien-3β-ol), ∆7,22-stigmastadienol = αspinasterol (stigmasta -7,22-dien-3β-ol), ∆7,22,25-stigmastatrienol (stigmasta -7, 22, 25trien-3β- ol), Δ7-ergostenol (27-methylcholest-7-en-3β-ol) and ∆7,24,28-stigmastadienol = ∆7-avenasterol (stigmasta-7, 24 (28) -dien-3β-ol) Bombardelli and Morazzoni 1997)

Squalene

Squalene (39-46%) is the characteristic constituent of the unsaponifiable fraction of the oil seeds. It can be used as a marker for the differentiation of oils obtained from other seeds (Bombardelli and Morazzoni 1997).

Triterpenoid: including 0.08-0.2% of multiflorane p-aminobenzoates (7-epi zucchini factor A and debenzoyl zucchini factor B ) (Appendino et al. 1999; 2000).

Sesquiterpenoids

The following monocyclic sesquiterpenoids have also been isolated: oxycerotic acid, (+) abscisic acid, (+) – 2-trans -abscisic acid, (+) – dehydrovomifoliol and (+) vomifoliol (Bombardelli and Morazzoni 1997)

Carotenoids 15 ppm (Vogel 1978), mainly lutein (50%) and β carotene (10-12%) with smaller amounts of cryptoxanthin and various other carotenoids (Bombardelli and Morazzoni 1997; Murkovic et al. 2002; Azevedo-Meleiro and Rodriquez -Amaya 2007).

Minerals

Particularly phosphorus, potassium, magnesium, calcium, iron, zinc and trace elements

(Mansour et al. 1993). Selenium is of particular importance as its content ranges between 0.08 and 0.4 μg/g, one of the highest values found in plants (Bombardelli and Morazzoni 1997). The pumpkin seeds contained relatively large amounts of potassium (5-90 μg/g dry weight) and chromium (approximately 3 μg/g dry weight). However, the sodium content of pumpkin seeds was low (6.9 μg/g dry weight).

Proteins and amino acids

Proteins are abundantly present in the seeds (31% – 51%) (Bombardelli and Morazzoni 1997; Bradley 2006; Glew et al. 2006).

Carbohydrates content is between 6% and 10% (Bombardelli and Morazzoni 1997).

Vitamins

In pumpkin seed of Cucurbita pepo Kakai 35 are present B group vitamins: thiamine 6.89; riboflavin 2.47; niacin 61.43; pyridoxine 4.92; pantothenic acid 4.95 (mg/kg) (Mansour et al. 1993).

Lignans

Secoisolariciresinol and lariciresinol were identified in pumpkin seeds (Sicilia et al. 2003

Cucurbita has been traditionally used as diuretic and anthelmintic, and as taenifugium remedy in Europe since medieval time. Pumpkin was mentioned in the writings of Hippokrates, Dioskurides, Lonicerus (1564), Fuchs (1523), Matthiolus (1626), (according to Madaus “Lehrbuch der Biologischen Heilmittel”, 1938).

Images of cucurbit have been decorating the Roman villa Farnesina since 1515.-1518. At that time , the first paintings of the species of New World cucurbits were known in Europe. Traditionally pumpkin seeds were used as anthelmintic, taeniacide remedy. Their contemporary use in functional disorders of the bladder with micturition difficulties is substantiated by empirical experience

In vivo preclinical pharmacology

Influence on urinary functions

The effects of an unspecified water-soluble extract of pumpkin seeds and soybean germ extract on in -bladder pressure (cystometrogram) and urination frequency of male rats were tested (Hata et al. 2005). Pumpkin seed water -soluble extract (250 mg/kg) compared to control solvent (1% dimethyl sulfoxide diluted in sterile physiological saline) and soybean germ extract significantly increased bladder volume, decreased urination frequency and increased urination delay index. According to the authors, the observed effects of the relaxation of the bladder and decrease of in -bladder pressure are related to the increased productions of NO via the arginine/NO pathway. Arginine is present in the pumpkin seed extract in two -fold the concentrations of other amino acids. It was suggested that arginine/NO metabolism, independently of adrenaline and acetylcholine, is involved in relaxation of urination muscle at a stage of full bladder (Andersson and Wein 2004.

Influence on prostate gland

Pumpkin seeds alleviated the signs of experimentally induced BPH in rats such as decrease of protein binding prostate (PBP) levels, size of the prostate and improved the histology of testis. Pumpkin seeds given orally (2.5, 5 and 10% in a diet) dose-dependently inhibited citral-induced hyperplasia of the prostate, especially at high concentration seed dose (10%, p<0.02) (Abdel -Rahman 2006).

In experimental model of hyperplasia of the prostate in rats, Gossell-Williams et al. (2006) tested the therapeutic effects of pumpkin seed oil. Hyperplasia of prostate gland was induced by a subcutaneous injection of testosterone: 0.3 mg/100 g of body weight (b.w.) /day for 20 days. The 1st group of tested rats received simultaneous administration of testosterone and pumpkin oil. The 2nd group received simultaneous administration of testosterone and corn oil for 20 days, the 3rd group only corn oil for 20 days. Pumpkin seed oil was administered in doses of 2.0 or 4.0 mg/100 g of body Weight for 20 days.

After autopsy on day 21, the prostate of each rat was weighed, and the prostate size ratio (weight of prostate/b .w. of the rat) was established. Testosterone significantly increased prostate size ratio (p<0.05) and this increase was significantly inhibited by treatment with pumpkin seed oil at 4.0 mg/100 g b.w.

Tsai et al. (2006) tested pumpkin seed oil efficacy for 14 days, in experiments performed in rats on the model of prostatic growth induced by subcutaneous daily injection of testosterone (1.25 mg/kg/day) together with prazosin (30 μg/kg/day) (T-P). Pumpkin seed oil (PSO) (2.5 ml/kg/day) extracted from pumpkin seeds was administered concomitantly together with T-P. As compared with T-P alone group, the T-P group treated with PSO had significant lower weight ratio for ventral prostate (p=0.01) and lower protein levels within ventral lobe and dorsolateral lobe (p=0.03 and p=0.003, respectively

Urodynamic effects

The urethral and bladder pressure were determined in anesthetized rabbits before and after administration of the extracts of pumpkin oil, n-butanol and ether fractions . The dose injected daily for 7 days was equivalent to 45 g of pumpkin seed. The bladder pressure was measured before and at 30, 60, 120 and 180 minutes after injection. Statistically significant (p<0.05) decrease of bladder and urethral pressure was registered only after oil fraction injections.

The decrease of bladder pressure was 7.6 mm Hg (p<0.001) with maximum effect after 2 hours, and urethral pressure 5.4 mg Hg (p<0.01as compared to pre-treatment values. The administration of the n-butanol and ether fraction had no significant influence (Zhang et al.

1994).

Anti-inflammatory activity

Fahim et al. (1995) described an anti-inflammatory activity of pumpkin seed oil administered in intramuscular injection in experimental arthritis in male Sprague -Dawley rats. Experimental arthritis was induced by inoculation of Freund’s complete adjuvant to the subplantar surface of the hindpaw. Pumpkin seed oil (100 mg/kg b.w.) was administered for 7 days before the adjuvant injection and then up to 22nd day afterwards.

For comparison, other groups of rats received in the same order: indomethacin (2 mg/kg b.w.), pumpkin seed oil + indomethacin and control group: 1% Tween 80. Both pumpkin seed oil and indomethacin were suspended in 1% Tween 80. During the acute and chronic phase of inflammation blood samples were collected for determination of blood glutation

(GSH), plasma total proteins, albumin serum sulfhydryl group (SH-gps), ceruloplasmin (CP) and lysosomal marker –N-acetyl-β-D-glucosaminidase (NAG). After completing the experiment, liver samples were used for determination of glucose-6-phosphate dehydrogenase (G-6-P DH) activity and protein content of liver homogenates was established. Adjuvant inoculation resulted in decrease of serum SH -gps, with an increase of serum CP reduction of blood glutathione and total proteins and albumins levels. Liver G6-P DH activity was markedly increased. The treatment with pumpkin seed oil resulted in normalization of altered parameters, notably in chronic phase, except serum NAG influence.

Pumpkin oil administration inhibited paw oedema during the chronic phase inabout 44% as compare Pumpkin oil administration inhibited paw oedema during the chronic phase in about 44% as compared to the control untreated group. It reduced also liver G-6-P DH activity to almost 50% of the arthritic groups’level. No potentiation of the antiinflammatory effects of indomethacin combined with pumpkin seeds oil was observed

Hepatoprotective activity

Protective effects of pumpkin seed protein isolate against live r damage induced by CCl4  in low -protein fed rats was estimated by Nkosi et al. (2005). The dried dehusked and defatted powder of Cucurbita pepo seeds was suspended in distilled water (pH 10) and filtered. The filtrate was centrifuged; the pH adjusted to 5 and the residue was freeze- dried. Experimental groups of Sprague-Dawley rats were kept on low -protein diet for 5 days and underwent experimental hepatic injury with IV injection of CCl4 . Two hours after the CCl4 administration, one group of rats received 1 ml/kg b.w. of the pumpkin seed protein isolate in saline (20g/100 ml) by gavage. Autopsy was performed at 2, 24, 48 and 72 hours after the CCl4 intoxication and plasma samples were tested. The significant increase of activity levels of lactate dehydrogenase (LD), alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) in plasma samples was registered in control intoxicated rats. Pumpkin seed protein isolate administration significantly reduced the level of LD, ALT, AST and ALP at all intervals tested (p<0.005). Neutralization of the effects of CCl4 was registered for LD 48 hours after treatment, and for AST, ALT and ALP 72 hours after intoxication.

Antiparasitic activity

The dried powdered seeds of Cucurbita maxima were extracted with 80% of ethanol, the filtrate was evaporated and the residue (PE) was dissolved in distilled water (concentration 500 mg/ml) (Lahon et al. 1978). The amino acid cucurbitine, isolated from Cucurbita moschata Duch. was administered orally to mice for 28 days since they were infected with 58-62 cercariae of Schistosoma japonicum (Shuhwa et al. 1962). Cucurbitine at the doses of 100, 200, 300, 400 and 500 mg/kg/day for 4 weeks, starting from the day of exposure infection, resulted in decreased average number of worm s (26.4, 25.3, 18.0, 11.8, 8.8 respectively) while that of the control infected group was 34.0 (p<005). When cucurbitine was administered 2-3 weeks after exposure, the prophylactic effect of the treatment was not shown.  Cucurbitine administration in the dose of 300 -400 mg resulted with a worm reduction rate of about 50% and retardation of their development Oral administration of powdered pumpkin seeds was tested in experimental infection of nodular worm (Oesophagostomum spp .) in piglets (Mägi et al. 2005). Groups of 4 crossbred nematode free piglets of both sexes with an average weight of 13.3 kg were inoculated per os by syringe with 5,000 larvae of Oesophagostomum spp. Each group received 5 g per kg of body weight of some medicinal plants, pumpkin seeds included, three times at weekly intervals after start of patency. For comparison, one piglet group was treated with 1% ivermectin. The test was terminated with autopsy and the worms were recovered from intestine by the agar -gel migration technique, counted and identified.

The pumpkin seeds treatment demonstrated the nematicidal effect as the number of worms recovered and excreted eggs was significantly lower compared to control untreated pigs. Moreover, therapeutic effects of pumpkin seeds were better than ivermectin administration Mahmoud et al (2002) described therapy after experimental infection with the trematode Heterophyes

Heterophyes in dog puppies with decoctions of pumpkin seeds and areca nuts. Decoctions of pumpkin seeds were prepared by boiling 10 g of grounded seeds in 15 ml of water for about an hour, decoction s of areca nuts by boiling 5 g in 15 ml of water. Both decoctions were orally administered, daily for 2 weeks after start of the infection (extract of pumpkin seeds: 10 g, extract of areca nuts: 5 g). In the group receiving an extract of pumpkin seeds, the deformation of eggs started on the 4 th day of treatment, however the complete destruction of eggs and eradication of adult worm s were acquired with combined extract therapy.

References

  • Abdel-Rahman MK. Effect of pumpkin seed (Cucurbita pepo L.) diets on benign prostatic hyperplasia (BPH): chemical and morphometric evaluation in rats. World J Chem 2006, 1(1):33 -40
  • Akihisa T, Shimizu N, Ghosh P, Thakur S, Rosenstein FU, Tamura RT, Matsumoto T. Sterols of the Cucurbitaceae. Phytochemistry 1987, 26:1693-1700
  • Andersson KE, Wein AJ. Pharmacology of the Lower Urinary Tract: Basis for Current and Future Treatments of Urinary Incontinence. Pharmacol Rev 2004, 56:581-631
  • Appendino G, Jakupovic J. Belloro E, Marchesini A. Multiflorane triterpenoid esters from pumpkin. An unexpected extrafolic source of PABA. Phytochemistry 1999, 51:1021-1026
  • Barbieri L, Polito L, Bolognesi A, Ciani M, Pelosi E, Farini V et al . Ribosome -inactivating proteins in edible plants and purification and characterization of a new ribosome inactivating protein from Cucurbita moschata .Biochim Biophys Acta 2006, 1760:783-792
  • Bastić M, Bastić LJ, Jovanović JA, Spiteller G. Sterols in pumpkin seed oil. J Am Oil Chem Soc 1977, 54:525-527
  • Bruneton J. Pharmacognosy, Phytochemistry, Medicinal Plants Lavoisier Publishing Inc, Intercept Ltd, London New York 1995
  • Duke JA. CRC Handbook of Medicinal Herbs. CRC Press, Inc. Boca Raton. London, New York, Washington 2000
  • Gruenwald J, Brendler T, Jaenicke C (Ed.). PDR for Herbal Medicines. Cucurbita pepo. Thomson Medical Economics, Montvale NJ. 2000
  • Madaus G. Lehrbuch der Biologischen Heilmittel. Cucurbita pepo. Kürbis. Cucurbitaceae.
  • Georg Thieme Verlag,  Leipzig 1938. Available at: http://www.henriettesherbal.com/eclectic/madaus/cucurbita.htm
  • Martindale. The Complete Drug Reference. The Pharmaceutical Press 2007
  • Tsaknis J, Lalas S, Lazos ES. Characterization of crude and purified pumpkin seed oil, Grasas & Aceites 1997, 48:267-272
  • Tsai Y -S, Tong Y -C, Cheng J -T, Lee C-Ho. Pumpkin seed oil and phytosterol -F can block testosterone/prazosin-induced prostate growth in rats. Urol Int 2006, 77:269-274
  • WHO Monographs on Selected Medicinal Plants, Volume 4. Semen Cucurbitae. World Health Organization, Geneva 2009, 83-91
  • Wichtl M. Cucurbitae semen. Pumpkin seed. In: Herbal Drugs and Phytopharma ceuticals.
  • Medpharm GmbH Scientific Publisher, CRC Press, Stuttgart: 2004, 170-172

Valerian

Valeriana officinalis L., radix and Valeriana officinalis L., aetheroleum Valerianae radix has been included into collections of monographs: in European Pharmacopoeia, British

Pharmacopeia (BP, 2015), United States Pharmacopoeia-NF (USP, 2014)

Definitions in the European Pharmacopoeia: Valerianae radix, monograph 07/2015:0453 Dried, whole or fragmented underground parts of Valeriana officinalis L. s.l., including the rhizome surrounded by the roots and stolons. content:  essential oil: minimum 4 ml/kg (dried drug), sesquiterpenic acids: minimum 0.17 per cent m/m, expressed as valerenic acid (C15H22O2; Mr234.3) (dried drug)

According to Wichtl (2009) the most important constituents of the plant are:

  • essential oil: 0.3-2.1% (according to Ph. Eur. not less than 0.4% and 0.3% for the whole drug or cut drug, respectively) with variable composition, depending on the origin, consisting of mono- and sesquiterpens; predominant components are bornyl acetate, myrtenyl-isovalerianate and –acetate; camphene, myrtenol, borneol are also found; important sesquiterpens are valerianol, valeranon, α-kessylacetate, βeudesmol, valerenal, tamariscene and the pacifigorgianes heavily volatile sesquiterpenic acids: (according to Ph. Eur. not less than 0.17%, expressed as valerenic acid) such as valerenic, acetoxyvalerenic acids and 3,4epoxyvalerenic acid, which have been considered species-specific marker compounds but based on recent studies have also been identified in non-aggregate species of officinalis
  • valepotriate: 0.1-2% main components valtrate and isovaltrate, in addition to dihydrovaltrate, IVHD-valtrate and the glycoside of valerosidatum; valepotriate are very unstable, their degradation products are e.g. baldrinal, homobaldrinal, valeric and isovaleric acid
  • small amonts of lignans (0.2%), e.g. 8-hydroxypinoresimol, its diglucoside, berchemol glucoside massoresionglucosid and 4’glucosyl-9-0-(6’’desoxysaccarosyl)-olivil
  • traces of alkaloids do occur (probably artefacts as a result from drying) such as actinidine, valerianine and the “main” valeriana-alkaloid, a pyridine derivative belonging to the monoterpene alkaloids
  • free amino acids like arginine, alanine, glutamine, GABA
  • flavonoids such as 8-methylapigenin, 2S-hesperidin, linarin
  • starch, various carbohydrates (glucose, fructose, saccharose, raffinose), phenol carbon acids and free fatty acids.

Therapeutic use of valerian root and its preparations probably goes back to the ancient Greeks and Romans who used a valerian-like herb for treatment of conditions for which therapists would use bitter and aromatic roots today. Dioscurides for example (Materia Medica 50-70 AD) used the dry root for warming, as a urinary tract remedy, for menstrual cramps and for liver diseases (Madaus, 1976). Since 18th century has been used as monotherapy in the modern phytotherapy in Europe

Chamisso (1781-1831) describes valerian root as antispasmodic, effective against worm diseases and as strengthening. Information on preparations and dose recommendations is not available (according to Benedum et al. 2000).

Vietz (1800) recommends valerian root for all spasmodic and convulsive diseases, for epilepsy, hysteric attacks, worm diseases and against nervous fever. Information on preparations and dose recommendations is not available (according to Benedum et al. 2000).

Hager (1873/74) refers to the use of valerian root for cramps, epilepsy, worm diseases and hysterical ailments. Information on preparations and dose recommendations is not available (according to Benedum et al., 2000.) The 25th edition of Martindale’s Extra Pharmacopoeia (1967) recommends the following uses of herbal preparations made from the dried rhizome and roots: indications: hysteria and other nervous condition, carminative List et al. (1979) list the following indications for valerian root mild sedative in nervous exhaustion, sleeplessness, mental overwork, nervous heart ailments, headache, neurasthenia, hysteria; as antispasmodic for stomach cramps, colics etc.

Primary pharmacodynamics

GABA (γ-aminobutyric acid) is an inhibitory neurotransmitter of the central nervous system that reduces nerve impulse transmission between neurons through the hyperpolarization of postsynaptic membranes and the reduction of neurotransmitter release into the synapse through presynaptic G-protein coupled receptor inhibition of voltage gattered Ca2+mechanisms (Jacob et al., 2008; Markwardt & Overstreet-Wadiche, 2008). Due to the essential inhibitory functions of GABA, the interest has focused on possible interactions of valerian root with the GABAA-messenger system.

in vivo activities in animals

Aqueous and aqueous-ethanolic valerian root extracts

Valerian tincture (1:5, no further information) reduced spontaneous motility after i.p. administration of the valerian tincture in mice (3.4625 ml/kg bw, corresponding to 0.6925 g herbal substance/kg bw) (Torrent et al., 1972).

Effects on spontaneous motility, thiopental sleeping-time and pentetrazol-induced toxicity were tested on mice by administering a commercially available aqueous dry valerian root extract (DER 5-6:1, extraction solvent: water). In spontaneous motility tests, doses of 20 and 200 mg/kg diminished the motility moderately, while in control animals 5 mg and 25 mg of diazepam resulted in substantial reductions in motility shortly after administration. The extract increased thiopental-induced sleeping time by factors of 1.6 at 2 mg/kg (p<0.01) and 7.6 at 200 mg/kg (p<0.01) compared to a factor of 4.7 for chlorpromazine at 4 mg/kg (Leuschner et al., 1993).

An aqueous-ethanolic dry extract (DER 4:1, extraction solvent: ethanol 70% V/V) administered i.p. to male mice, was assessed for possible neuropharmacological activity. At doses of up to 100 mg/kg the extract did not produce sedation nor tranquillization, since no modifications of spontaneous motility, nociception or body temperature and no palpebral ptosis were observed, whereas diazepam at doses of up to 2 mg/kg clearly reduced spontaneous motility, lowered body temperature and produced a weak ptosis. However, the extract showed a significant prolongation of thiopental-induced anaesthesia (p<0.05) with 100 mg/kg of extract (Hiller & Zetler, 1996).

Cats with implanted electrodes showed no changes in their EEGs following oral administration of 100 or 250 mg of valerian root extract (no further information) per kg

b.w. The muscle tonus was reduced in 30-40% of the animals (Holm et al., 1980).

An aqueous-ethanolic (extraction solvent ethanol 70% V/V) valerian root extract had an anxiolytic effect in the elevated plus-maze test in male Spraque Dawley rats after oral administration in the magnitude of ipsapirone, a 5-HT1A-receptor agonist (Hiller & Kato, 1996). After single administration of 5, 25 and 100 mg/kg, the extract showed a distinct anxiolytic-like effect while the observed effect was not significant for 1 mg/kg and lower doses. Remarkably, a moderate anxiolytic-like effect was maintained during subchronic administration over 5 days only for low doses of 1 mg/kg.

Sichardt et al. (2007) compared  the action of a methanol (M-E; extraction solvent: methanol 45% m/m; valerenic acid 0.332%; concentration 10 mg/ml), ethanol (E-E; extraction solvent: ethanol 62.4% m/m; valerenic acid 0.127%; concentration 10 mg/ml ) and an extract (EA-E; marc macerated with ethylacetate; valerenic acid 2.956%; concentration 10 mg/ml) from valerian roots on postsynaptic potentials (PSPs) in cortical neurons. Intracellular recordings were performed in rat brain slice preparations containing pyramidal cells of the cingulate cortex. PSPs were induced by electrical field stimulation. The M-E induced strong inhibition in the concentration range 0.1-15 mg/ml, whereas the E-E (1-10 mg/ml) did not influence significantly the PSPs. The maximum inhibition induced by the M-E was completely antagonized by 1,3-dipropyl-8cyclopentylxanthine (DPCPX, 0.1 µM), an antagonist on the adenosine A(1) receptor. Contrary to the M-E, the EA-E (10 mg/ml) induced an increase of the PSPs, which were completely blocked by the GABAA receptor antagonist picrotoxin (100 µm).

The study of Chow et al. (2011) investigated the sedative effects of a valerian root extract (DER: 3-6:1; extraction solvent: 70% ethanol; 91% native extract). Locomotor activity and core body temperature were recorded in male mice using radiotelemetry. The extract and some of its single constituents, valerenic acid, linarin, and apigenin, were tested for effects on locomotion and body temperature over 180 min after oral administration. The extract was tested in a dose range of 250-1000 mg/kg, and only the highest dose showed a mild short-term sedative effect with reduced locomotor activity between 66-78 min after administration. Paradoxically, an increased activity was observed after 150 min after gavage. A dose of 1 mg/kg valerenic acid produced an intermittent stimulation of activity.

However, a mild short-term sedative effect was found for linarin at 12 mg/kg and apigenin at 1.5 mg/kg. Considering the cumulative locomotor activity over the observation period of 180 min, the authors concluded that neither the extract nor one of the compounds had considerable sedative effects. More precisely, the observed short-term changes in activity pattern indicate that valerian extract as well as the flavonoids linarin and apigenin are rather effective to reduce sleep latency than to act as a sleep-maintaining agent.

Sesquiterpenoids

The i.p. application of sesquiterpenoid compounds like valerenic acid, valerenal and valeranone isolated from the essential oil of valerian root revealed a sedative and muscle relaxant acitivity. In addition, valerenic acid and valeranone were found to prolong the barbiturate induced sleeping time (Hendriks et al., 1981, 1985; Rücker et al., 1978; Torrent et al., 1972; Hänsel et al., 1994).

Valepotriates

Valerenic acid and valerenol exerted anxiolytic activity with high potencies in the elevated plus maze and the light/dark choice test in wild type mice. In beta3 (N265M) pointmutated mice the anxiolytic activity of valerenic acid was absent. Thus, neurons expressing beta3 containing GABAA-receptors seems to be a major cellular substrate for the anxiolytic action of valerian extracts (Benke et al., 2009).

Nam et al. (2013) investigated the effects of extract from valerian root (DER 4:1; no further information) and its major component, valerenic acid on memory function, cell proliferation, neuroblast differentiation, serum corticosterone, and lipid peroxidation in adult and aged mice. For the aging model, D-galactose (100 mg/kg) was administered s.c. to 6-week-old male mice for 10 weeks. At 13 weeks of age, valerian root extract (100 mg/kg) or valerenic acid (340 μg/kg) were administered orally to control and Dgalactose-treated mice for 3 weeks. The dosage of valerenic acid (340 μg/kg) was determined by the content of valerenic acid in valerian root extract (3.401±0.066 mg/g) measured by HPLC. The administration of valerian root extract and valerenic acid significantly improved the preferential exploration of new objects in novel object recognition test and the escape latency, swimming speeds, platform crossings, and spatial preference for the target quadrant in Morris water maze test compared to the D-galactosetreated mice. Cell proliferation and neuroblast differentiation were significantly decreased, while serum corticosterone level and lipid peroxidation in hippocampus were significantly increased in the D-galactose-treated group compared to that in the control group. The administration of valerian root extract significantly ameliorated these changes in the dentate gyrus of both control and D-galactose-treated groups. In addition, valerenic acid also mitigated the D-galactose-induced reduction of these changes.

References

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  • Hänsel R, Sticher O, Steinegger G. (editors) Pharmakognosie – Phytopharmazie, 6th ed. Springer Verlag, Berlin Heidelberg New York 1999, 457-464 [German]
  • Hendriks H, Bos R, Allersma DP, Malingré TM, Koster AS. Pharmacological screening of valerenal and some other components of essential oil of Valeriana officinalis. Planta Med. 1981, 42: 62-68
  • Hendriks H, Bos R, Woerdenbag HJ, Koster AS. Central nervous depressant activity of valerenic acid in the mouse. Planta Med. 1985, 51: 28-31
  • Holm E, Kowollik H, Reinecke A, v. Henning GE, Behne F, Scherer HD. Vergleichende neurophysiologische Untersuchungen mit Valtratum/Isovaltratum und Extractum Valerianae an Katzen. Med. Welt. 1980, 31(26): 982-990 [German]
  • Madaus G. Lehrbuch der biologischen Heilmittel. Vol. 2. Georg Olms Verlag, Hildesheim New York 1976 (reprint of Madaus G. Lehrbuch der biologischen Heilmittel. Georg Thieme
  • Verlag, Leipzig, 1938), 1267-1278 [German] Martindale The Extra Pharmacopoeia. 25th ed. Percy Lund, Humphries & Co. Ltd, London Bradford 1967, 2046-2047
  • Nam SM, Choi JH, Yoo DY, Kim W, Jung HY, Kim JW et al. Valeriana officinalis extract and its main component, valerenic acid, ameliorate D-galactose-induced reductions in memory, cell proliferation, and neuroblast differentiation by reducing corticosterone levels and lipid peroxidation. Exp Gerontol. 2013, 48(11): 1369-1377
  • Neamati A, Chaman F, Hosseini M, Boskabady MH. The effects of Valeriana officinalis L. hydro-alcoholic extract on depression like behavior in ovalbumin sensitized rats. J Pharm Bioallied Sci. 2014, 6(2): 97-103
  • Rücker G, Tautges J, Sieck A, Wenzl H, Graf E. Untersuchungen zur Isolierung und pharmakodynamischen Aktivität des Sesquiterpens Valeranon aus Nardostachys jatamansi DC. Arzneim.-Forsch./Drug Res. 1978; 28(I) (1): 7-13 [German]
  • Sichardt K, Vissiennon Z, Koetter U, Brattström A, Nieber K. Modulation of postsynaptic potentials in rat cortical neurons by valerian extracts macerated with different alcohols: involvement of adenosine A(1)- and GABA(A)-receptors. Phytother Res. 2007, 21(10): 932-937
  • Torrent MT, Iglesias J, Adzet T. Valoración experimental de la actividad sedante de la tintura de Valeriana officinalis L. Circular Farmacéutica. 1972, 30:107-112 [Spanish]
  • WHO (World Health Organistaion). WHO monographs on selected medicinal plants. Vol.1. Radix Valeriana. World Health Organization. Geneva 1999, 267-276
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Californian poppy

Eschscholzia californica Cham. is a component part of the French Pharmacopoeia. The following monograph exists:

Eschscholzia (poudre d’), French Pharmacopoeia, 10th ed. (Fr.Ph. Jan 1996): Dried and powdered flowering aerial parts of Eschscholzia californica Cham. Content: not less than 0.50 per cent and not more than 1.20 per cent of total alkaloids, expressed as californidine (C20H20NO4+; MT 338.4) (dried drug).

Eschscholzia californica Cham. (Fam. Papaveraceae) is a perennial and annual plant growing approx 30 cm high with alternately branching glaucous blue-green foliage. The flowers are solitary on long stems, silky-textured, with four petals, each petal 2–6 cm long and broad; their color ranges from yellow to orange. The plant is prolific, with numerous black or dark brown colored seeds held in the center of the flower within slender, ribbed single celled seed capsules (3-9cm long) (Bruneton, 1998).

Both aerial parts and roots contain alkaloids, the latter being richer than the former (up to 1.6% alkaloids) (Fleurentin et al., 1996).

Synonyms: California poppy

Constituents  

Alkaloids: 0.50-1.20 per cent of total alkaloids, expressed as californidine; six different groups of alkaloids have been described.

Pavin alkaloids in the aerial parts (most abundant and characteristic of this Genus):

Aporphine alkaloids: laurotetamine and N-methyl laurotetamine are present in the whole plant, glaucine can be found within the aerial parts

Protopine alkaloids: protopine, cryptopine and α-cryptopine are found in the whole plant, leaves and stem (Fleurentin et al., 1996).

Benzo-phenanthridin alkaloids: Chelirubine, chelidonine and homochelidonine are present in the whole plant, chelilutine being found in aerial parts.

The group of benzo-C-phenanthridin alkaloids, and mainly sanguinarine and chelerythrine, are not present in aerial parts (Fleurentin et al., 1996). 

Others: Flowers contain rutin and a purple-red pigment eschscholtz-xanthin

California poppy is a traditional medicinal plant from North-american Indian population, nowadays being used for its mild analgesic and sedative properties (and as the state flower of California) (Baldacci, 1990; Bocek, 1984; Mills and Bone, 2000; Duke 2001) without the dangers attending opiates. This species is native to California (USA) and North area of Mexico and has perfectly been adapted to several European countries where it is frequently cultivated in ornamental gardens. Traditionally used by the rural population of western USA as analgesic and sedative (Fleurentin, 1993).

A recent review on Plant-based medicines for anxiety disorders (Sarris et al., 2013) includes Eschscholzia californica as a plant with preclinical evidence of anxiolytic activity.

In vivo studies

The intraperitoneal administration of the aqueous extract of the plant at 25mg/kg in mice exerted an anxiolytic action, as proved by changes in behavioural parameters; at higher levels, the effect became more sedative (Mills and Bone, 2000).

The study performed by Rolland et al., (1991) intended to validate the traditional sedative indications of E. californica by pharmacological investigations. Thus, several doses of the aqueous extract from aerial parts of the plant (from 25 to 400mg/kg) were tested to determine the possible sedative and/or anxiolytic effects on the behaviour of mice subjected to several experimental situations. Naive male mice (Swiss) weighing 30-35g were used for behavioural tests (two compartments test, sleep induction test, staircase test) while naïve male and female Swiss mice were used for acute toxicity determination.

With respect to the first experiment, E. californica induces a dose-dependent decrease of the number of rearing and the total locomotion from the dose of 100mg/kg while the novelty preference is significantly affected only from 200mg/kg. ED50 is estimated to be 151mg/kg for the locomotion reduction and 108mg/kg for the rearing reduction. In the sleep induction test, California poppy aqueous extract induced a dose-dependent sleep induction from the dose of 100mg/kg. The sleeping induction ED50 was estimated to be 106mg/kg. In the same conditions, dipotassium clorazepate as the reference compound, induced significantly and dose-dependent sleep in mice from the dose of 5mg/kg (50% of sleeping mice). The results obtained after the exposition of mice to the staircase test demonstrated that E. californica significantly decreased the number of steps climbed and the number of rearings affected by mice, from 200mg/kg (at lower dose of 25mg/kg, the effects were reversed). ED50 was estimated to be 254mg/kg for the rearings and 153mg/kg for the steps climbed; data obtained for the reference compound, dipotassium clorazepate, were a sedative ED50 of 13mg/kg and 23mg/kg for the rearings and the stpes climbed, respectively. Also E. californica significantly increased the time spent by mice in the lit box at the dose of 25mg/kg.

In relation to the acute toxicity test, the aqueous extract of E. californica did not induced the mortality up to the dose of 8mg/kg afer i.p. and per os administration. Animals did not show any toxic manifestation on the studied parameters. The evolution of body weigth was normal, despite a small decrease in the first four hours when eyes were closed, this effect corresponding to the sedative activity.

References

  • Bocek BR. 1984. Ethnobotany of Costanoan Indians, California, based on Collections by John P. Harrington. Economic Botany. 38(2): 240-255.
  • Baldacci R. 1990. Contribution à l’utilisation en médecine de ville d’Eschscholtzia californica. Phytotherapy 22: 31-33.
  • Blumenthal M, Busse WR, Goldberg A, Gruenwald J, Hall T, Riggins CW, Rister RS. 1998.
  • The complete German commission E monographs. Therapeutic guide to herbal medicines, American Botanical Council, Austin. Texas. USA. pp. 389
  • Bruneton J. Pharmacognosie. Phytochimie, plantes médicinales. Tec et Doc, 1999; 3e éditions. Paris. France. pp 745-746.
  • Duke JA. 2001. Handbook of Medicinal Herbs. CRC Press Boca Raton, Florida. USA. p. 184
  • Fleurentin J. 1993. Ethnopharmacologie et aliments: introduction au sujet et réflexions sur l’efficacité biologique. Actes du 2ème Colloque Européen d’Ethnopharmacologie et de la 11ème Conférence Internationale d’Ethnomédecine, Heidelberg, 24-27 mars 1993.
  • Fleurentin J, Mortier F, Younos C. 1996. Eschscholtzia, Eschscholtzia californica Cham. Papaveraceae, in Acupunture & Médicine Traditionnelle Chinoise. Phytothérapie & aromathérapie. Homéopathie. Ed. Frison-Roche, Paris. France. pp. 95-100.
  • French Pharmacopoeia monograph “Eschscholtzia (Parties Aeriennes Fleuries”. 10th ed., January 1996
  • Rolland A, Fleurentin J, Lanhers MC, Younos C, Misslin R, Mortier F, Pelt JM. 1991. Behavioural effects of the American traditional plant Eschscholzia californica: sedative and anxiolytic properties. Planta Med. 57(3): 212-216.
  • Rolland A, Fleurentin J, Lanhers MC, Younos C, Misslin R, Mortier F. 2001. Neurophysiological effects of an extract of Eschscholzia californica Cham. (Papaveraceae). Phytother Res. 15(5): 377-381.
  • Sarris J, McIntyre E, Camfield DA. 2013. Plant-based medicines for anxiety disorders, Part 1: a review of preclinical studies. CNS Drugs. 27(3): 207-219. doi: 10.1007/s40263-0130044-3.

Melatonin

Melatonin is a hormone produced in animals by the pineal gland and in plants under stress. Melatonin research has expanded rapidly, affecting an impressive enhancement in the understanding of its functions in plants and animals. However, far less focus has been directed to clarifying the nature of melatonin dose-response relationships. The characteristics of these dose responses are similar to those of the broad toxicological and pharmacological hormesis literature. Analysis suggests that melatonin, in coordination with the circadian rhythms, is involved in stress adaptive responses, and may act as a conditioning agent protecting organisms against subsequent health threats within an hormetic framework. Incorporation of melatonin-induced hormesis in research protocols has the potential to enhance the treatment of neuropsychiatric diseases, cancers, and other animal diseases, as well as protection against environmental stress and to increase plant productivity.

Melatonin was first discovered in connection to the mechanism by which some amphibians and reptiles change the color of their skin. In 1958, dermatology professor Aaron B. Lerner and colleagues at Yale University, in the hope that a substance from the pineal might be useful in treating skin diseases, isolated the hormone from bovine pineal gland extracts and named it melatonin. In the mid-70s Lynch et al. demonstrated that the production of melatonin exhibits a circadian rhythm in human pineal glands. The discovery that melatonin is an antioxidant was made in 1993. The first patent for its use as a low-dose sleep aid was granted to Richard Wurtman at MIT in 1995. Around the same time, the hormone got a lot of press as a possible treatment for many illnesses.

Female fertility irreversibly declines with aging and this, is primarily associated with the decreased quality and quantity of oocytes. To evaluate whether a long-term of melatonin treatment would improve the fertility of aged mice, different concentrations of melatonin (10-3, 10-5, 10-7 M) were supplemented into drinking water. Melatonin treatments improved the litter sizes of mice at the age of 24 weeks. Mice treated with 10-5 M melatonin had the largest litter size among other concentrations. At this optimal concentration, melatonin not only significantly increased the total number of oocytes but also their quality, having more oocytes with normal morphology that could generate more blastocyst after in vitro fertilization in melatonin (10-5 M) treated group than that in the controls.

Pineal melatonin is synthesized and secreted in close association with the light/dark cycle.

The temporal relationship between the nocturnal rise in melatonin secretion and the «opening of the sleep gate» (i.e., the increase in sleep propensity at the beginning of the night), coupled with the sleep-promoting effects of exogenous melatonin, suggest that melatonin is involved in the regulation of sleep. The sleep-promoting and sleep/wake rhythm regulating effects of melatonin are attributed to its action on MT(1) and MT(2) melatonin receptors present in the suprachiasmatic nucleus (SCN) of the hypothalamus. Animal experiments carried out in rats, cats, and monkeys have revealed that melatonin has the ability to reduce sleep onset time and increase sleep duration. However, clinical studies reveal inconsistent findings, with some of them reporting beneficial effects of melatonin on sleep, whereas in others only marginal effects are documented. Recently a prolonged-release 2-mg melatonin preparation (Circadin(TM)) was approved by the European Medicines Agency as a monotherapy for the short-term treatment of primary insomnia in patients who are aged 55 or above. Several melatonin derivatives have been shown to increase nonrapid eye movement (NREM) in rats and are of potential pharmacological importance been shown to improve sleep in depressed patients and to have an antidepressant efficacy that is partially attributed to its effects on sleep-regulating mechanisms.

Melatonin might constitute a useful tool for the treatment of an array of conditions in animals, such as feline uveitis.

References