Journal of Experimental and Integrative Medicine

www.jeim.org 10.5455/jeim.260416.rw.015 Mini Review DOI: A review on phyto-pharmacological potentials of Juniperus recurva

Abila Rashid1, Raja Amir Hassan Kuchay2

1Faculty of Pharmaceutical ABSTRACT Sciences, Shri Venkateshwara University, Gajraula, UP, India. Juniperus recurva is a small or spreading distributed from Afghanistan eastward through the Himalayas to 2School of Biosciences & southwest . It is predominant in woodlands of the wetter areas of Himalayas (altitude of 1800 m to 3900 m), with Biotechnology, CBS, BGSB University, moist and well-drained soils. It holds great potential as a medicinal . It has been traditionally used as anti-rheumatic, J&K, India. antiseptic, antispasmodic, antitoxic, aphrodisiac, astringent, carminative, rubificant and stomachic. However, very few authentic pharmacological studies on medicinal benefits of this plant have been conducted thus far. In a first of its kind, Address for correspondence: we review this plant for its chemical constituents, pharmacological activity and medicinal benefits. Keeping in view its Raja A.H Kuchay, success record as a traditional medicine to cure different ailments, it would be of interest to look for various bioactive School of Biosciences & natural products that might have therapeutic potential. Biotechnology, CBS, BGSB University, J&K, India. [email protected] Received: January 12, 2016 Accepted: April 26, 2016 Published: June 30, 2016 KEY WORDS: Juniperus recurva; Distribution; Pharmacological Activity.

INTRODUCTION survey. No limits were imposed in terms of author or type of publication. Juniperus L. (family ), a genus of evergreen aromatic or is distributed in temperate and OCCURRENCE AND MORPHOLOGY cold regions of northern Hemisphere [1]. Depending on taxonomic viewpoint, between 50 and 67 species of J. recurva grows from Pakistan to south west China and is are widely distributed throughout the northern Hemisphere, particularly common in Nepal and Kashmir [2–6]. It can from the Arctic, south to tropical Africa in the old world, be found as a large shrub in hilly region of Nepal. It can and to the mountains of central America, Pakistan, India also be tree having height of 6-20 m and a trunk up to 2 and China [2–6]. remain green for several years and m diameter and a broadly conical to rounded or irregular even after drying may remain on the shoots for indefinite crown. In Nepal it is widely spread in east side of Himalaya period [2–6]. Flowers are unisexual [2–6]. In some species region. In Kashmir it is found in Khilanmarg and on the flowers of both the sexes occur on the same plant [2–6]. All route from Sonamarg to Amarnath cave at an altitude of the species can be propagated through ; seeds retain 3000-4000 m. It mainly grows at altitude of 3000-4000 viability for several years when stored in cool dry place m. Two varieties of J. recurva have been named: J. recurva [2–6]. like open positions and chalky or limestone var. recurva found throughout the Himalayan range and J. soils [2–6]. Common Juniper species of Himalayan range recurva var. coxii mainly confined to the eastern Himalaya include: J. communis, J. indica, J. recurva and J. squamata [2, on high rainfall sites [2, 8]. However, a report has listed 7]. Juniperus recurva is commonly known as the ‘Drooping J. recurva var. squamata and J. recurva var. coxii as the two Juniper’ [2]. species of Himalayas [9]. J. recurva is monoecious or rarely dioecious. Trunk bark is METHODS generally light grayish brown or brown, crown conical or Authors carried out electronic searches in PubMed, Embase, broadly pyramidal. Branches are ascending in apical part of Medline Google Scholar, Scopus and Cochrane Library. plant and spreading toward base. Leaves are needle-like, 5-10 Authors screened the titles and abstracts independently to mm long, arranged in six ranks in alternating whorls of three. identify potential eligible articles. Then full-text articles cones are -like, globose to ovoid, 5-10 mm long and were retrieved if necessary. Authors used controlled 4-7 mm diameter, glossy blue-black, and contain one seed; vocabulary and text words to construct a highly sensitive they are mature in about 18 months. The male cones are 3-4 search strategy during the entire process of literature mm long, and shed their pollen in early spring [2].

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SCIENTIFIC CLASSIFICATION (1) Kingdom: Plantae (2) Division: Pinophyta (3) Class: Pinopsida (4) Order: (5) Family: Cupressaceae (6) Genus: Juniperus (7) Species: Juniperus recurva

THREATS In the IUCN (International Union for Conservation of Nature) Red List of Threatened species Juniperus recurva var. coxii has been put in the category of Near Threatened [10]. Having previously been listed as Vulnerable, it was considered that there is insufficient evidence to assume an overall reduction greater than 30%. In certain areas, e.g. Yunnan, deforestation has been cause of reduction in area of occupancy, but elsewhere the forest is still intact. It seems more appropriate to list this taxon as Near Threatened [10]. In its native countries J. recurva Fig. 1. Essential Oil Composition of J. recurva var. squamata [6,11] is used for timber as well as ornamental trees in gardens of monasteries and temples. The wood and foliage are burned alkaloids, steroids and terpenoids [12]. In a preliminary for incense in Buddhist temples [8]. In Myanmar (Burma) phytochemical screening of methanolic extract of six species the wood of large trees is used to make coffins. The drooping of Juniperus including J. recurva, the presence of alkaloids, form J. recurva var. coxii is a highly ornamental tree much tannins, flavonoids, terpenoids and steroids was reported valued and often planted in regions of Europe with a mild, [13]. The contents of phenolic compounds were highest in moist climate [8]. J. communis and the total flavonoid contents were highest in J. indica. For J. recurva total phenolic content in (equiv L-1) was 2.10 ± 0.13 and total flavonoid content (equiv L-1) CHEMICAL CONSTITUENTS was 0.890 ± 0.17 [13]. Amentoflavone was also isolated Analysis of the composition of the leaf oils of J. recurva from aerial parts of different Juniperus species including from India (Sikkim, Nepal) and its comparison to the J. recurva and authenticated by high performance liquid composition of J. recurva var. squamata from western chromatography (HPLC) and physicochemical parameters India has been reported [6]. The major components of J. [13]. recurva oil were -pinene, sabinene, -3-carene, limonene, Production of podophyllotoxin by an endophytic fungus terpinen-4-ol, -cadinene, -cadiene, elemol, cubenol, Fusarium oxysporum isolated from the medicinal plant J. α δ epi- -cadinol, epi- -muurolol, -cadinol and 4-epi-abietal recurva has been reported [14]. Further confirmation and γ δ [6]. Study on essential oils of leaves and twigs of Juniperus quantification of podophyllotoxin was performed by HPLC, α α α recurva var. squamata by gas liquid chromatography (GLC), liquid chromatography-mass spectrometry (LC-MS), and mass spectrometry (MS), and nuclear magnetic resonance LC-MS/MS [14]. Podophyllotoxin is a well-known naturally (NMR) spectroscopy has also been performed [11]. Main occurring aryltetralin lignan that occurs in few plant species constituents were monoterpene hydrocarbons (68%); that is used as a precursor for the chemical synthesis of the in addition, about 12% of sesquiterpene hydrocarbons, anticancer drugs like etoposide, teniposide and etopophose 9% of oxygenated sesquiterpenes, and 5% of oxygenated phosphate. The availiability of this lignan is becoming monoterpenes were identified. 4 -Hydroxygermacra-1, increasingly limited because of the scarce occurance of 5-diene, -oplopenone, oplopanone, 8-acetoxy elemol, and its natural sources and also because synthetic approaches manool were found as trace componentsβ (Figure 1) [11]. for its production are still commercially unacceptable [14]. β Recently a study was conducted on the phytochemical screening for the presence of different phytochemicals PHARMACOLOGICAL ACTIVITIES AND MEDICINAL like alkaloids, tannins, reducing sugar, coumarin, glycosides, quinones, steroids, terpenoids and saponin in USES the different plant parts of J. recurva [12]. Leaf and seed Essential oils and their components are widely used in fractions demonstrated differences in the presence of medicine as constituents of different medical products, various phytochemicals. Leaf fractions showed only the in the food industry as flavouring additives and also in presence of coumarins and quinones with the absence of cosmetics as fragrances and pharmaceutical industries [15, other mentioned phytochemicals. Seed fractions showed 16]. Essential oils are valuable natural products used as raw the presence of tannins, coumarin, glycosides, saponin, materials in many fields, including perfumes, cosmetics,

94 J Exp Integr Med ● 2016 ● Volume 6 ● Issue 2 Abila & Amir: Juniperus recurva Phyto-Pharmacological Activities aromatherapy, phototherapy, spices and nutrition [17]. leaf and seed extract fractions showed the concentration Also the essential oils are used in traditional medicine for dependence antimicrobial activity towards the bacteria used their antiseptic action. Juniper berry essential oil is found in with the highest ZOI being 17 mm for the J. recurva leaves soaps and in pomades with the aim of curing alopecia [18]. at 200 mg/ml [12]. Various other species of Juniperus have The oil is also irritating to microbes, so much so that it kills also been reported to have antimicrobial activity [27, 28]. many of them [19]. Juniper berries (mature female cones) Sensitivity of five pathogenic multi drug resistant bacteria have long been used as flavouring agents in foods and (Bacillus subtilis, Erwinia chrysanthemi, Escherichia coli, alcoholic beverages such as gin [20]. In herbal medicine, Agrobacterium tumefaciens and Xanthomonas phaseoli) was Juniper oil has been used as a carminative, diuretic and as a tested against the crude leaf organic extracts (methanol, steam inhalant in the management of bronchitis. It has also ethanol, chloroform and hexane) and aqueous extracts of been used in arthritis as well as antioxidant [21]. Berries a Kumaun Himalayan gymnospermous plant J. communis, are also recommended in cough, infantile tuberculosis employing disc diffusion method. All the extracts of J. and diabetes, whereas, ash of the bark is used for certain communis were found effective by showing a mark zone skin diseases [22, 23]. Mentioned below are few reported of inhibition except aqueous extract [27]. Antimicrobial pharmacological activities associated with J. recurva and its activity of Juniper essential oil has also been reported in different constituents. many other studies [29, 30].

Insecticidal Activity Antioxidant Activity Many insecticidal principles have been reported in the Furthermore, antioxidant (2,2-diphenyl-1-picrylhydrazyl; heartwood of J. recurva including thujopsene and 8-cedren- DPPH) assay of different plant-parts of J. recurva has 13-ol. Additional 12 sesquiterpenes were also detected also been investigated [12]. The results demonstrate that in the neutral fraction of ether extracts [24]. The active the selected plant extract fractions of J. recurva possess components from the heartwood of J. recurva were monitored significant antioxidant activities. Antioxidants compounds for the insecticidal activity against mosquito female adults scavenge free radicals such as peroxide, hydroperoxide or (Culex pipiens pallens) with topical application test [24]. lipid peroxyl and thus inhibit the oxidative mechanisms Essential oils of two Tibetan Junipers, J. saltuaria and that lead to degenerative diseases [12]. Antioxidant effects J. squamata var. fargesii have also been reported to have of J. communis essential oils has also been reported in vivo insecticidal activity against adult lace bugs [25]. [31]. The protective effect of methanolic extracts from different Xanthine Oxidase Inhibitory Activity species of Juniperus in different concentration has been reported against H2O2 induced oxidative damage in human Xanthine oxidase inhibitory activity of J. recurva has been erythrocytes and leucocytes [13]. Human blood cells are recently assessed [26]. The inhibitory effect of polyphenolic the key fighters against various oxidative stress diseases rich fractions of J. recurva was determined to find out the and remain the primary target for natural products and natural inhibitors of xanthine oxidase. Xanthine oxidase . These cells are continuously exposed to radicals is a cytoplasmic enzyme, converting hypoxanthine to and are unable to replace damaged components, especially xanthine and generates superoxides and uric acid. Gout polyunsaturated fatty acids chains from peroxidation [32]. attacks can be treated with the use of xanthine oxidase The measurements of different antioxidant parameters inhibitors. Allopurinol is generally used in the management like superoxide dismutase, glutathion peroxidase, lipid of gout and conditions associated with hyper-uricemia but peroxidase, catalase and glutathione levels were performed its adverse effect leads to GIT distress, hypersensitivity under each set of extracts including J. recurva. Isolated reactions and hepatitis. It was reported that the methanol fractions from all the six Juniperus species were reported fraction of J. recurva has more inhibition against xanthine as potential source of natural antioxidants for treatment oxidase activity in the samples treated with different and prevention of diseases in which oxidative stress takes concentrations of hypoxanthine on second, third, fourth place [13]. Antioxidant potential has also been reported and fifth weeks as compared to other plant extracts [26]. for essential oils of Juniper berries of different species of Junipers [33, 34]. Antimicrobial Activity Cytotoxic Activity Antimicrobial assay with the leaf and seed plant extracts of J. recurva by agar plate diffusion method has been In-vitro cytotoxicity assay with the leaf and seed plant reported [12]. Five antibiotics were used as standard extracts of J. recurva demonstrated its anticancer properties drugs. Six different bacteria Bacillus cereus, Bacillus against MCF7 (Breast Cancer) cell lines [12]. The thuringiensis, Escherichia coli, Pseudomonas aeruginosa, cytotoxic effect or the inhibitory effect of these extracts was Bacillus subtilis and Proteus mirabilis were used for the compared with standard (Dexorubicin). The percentage assay. The antimicrobial activity was assayed by measuring inhibition was reported to be concentration dependent. It the diameter Zone of Inhibition in millimetres (ZOI) of was suggested that isolated pure compounds with bioassay different extracts on the agar disc plate. Both the J. recurva guided extraction, could possibly get the higher inhibitory

J Exp Integr Med ● 2016 ● Volume 6 ● Issue 2 95 Abila & Amir: Juniperus recurva Phyto-Pharmacological Activities effect against MCF7 cells from the plant J. recurva. REFERENCES Essential oils of few other species of Juniperus have also 1. Hooker JD. The flora of British India. Publ, Bishen Singh Mahendra been reported to have cytotoxic activity [35–37]. Pal Singh, Dehradun, 1999. 2. Adams RP. Junipers of the world: The genus Juniperus. Trafford Publ, Canada, Victoria BC, 2004. Other Uses 3. Farjon A. World checklist and bibliography of . Kew, 2001. 4. Farjon A. Monograph of Cupressaceae and Sciadopitys. Royal Various parts of J. recurva are tradition used as anti- Botanic Gardens, Kew, 2005. 5. Mao K, Hao G, Liu J, Adams RP, Milne RI. Diversification and rheumatic, anti-septic, anti-spasmodic, astringent, biogeography of Juniperus (Cupressaceae): variable diversification carminative, depurative, stimulant, stomachic, tonic, and rates and multiple intercontinental dispersals. New Phytologist in aroma industry. One of the most ancient rituals of the 2010; 188(1):254–72. 6. Adams P, Thappa RK, Agrawal SG, Kapahi BK, Srivastava TN, Himalayan region is the burning of J. recurva on mountain Chaudhary RP. The leaf essential oil of Juniperus recurva Buch.- tops to purify and sanctify the area. J. recurva has also Ham. Ex D. Don from India and Nepal compared with Juniperus been reported to be useful in the treatment of various recurva var. squamata (D. Don) Parl. J Essent Oil Res 1998; 10(1):21–4. ailments including fever, headache, cough, cold and kidney 7. Lohani H, Haider SZ, Chauhan NK, Mohan M. Essential oil problems. High F (informant consensus factor) value composition of leaves and berries of Juniperus communis and IC Juniperus indica from Uttarakhand Himalaya. J Med Aromat Plant was obtained for these ailments, indicating that the J. Sci 2010; 32(3):199–201. recurva traditionally used to treat these ailments is worth 8. Farjon A. A handbook of the World’s conifers. Koninklijke Brill, searching for bioactive compounds [38]. Summary of its Leiden, 2010. 9. Gulati BC, Shawi AS, Garn SN, Channa GN. Essential pharmacological activities and traditional uses is given in oil of Juniperus recurva var squamata and other oils of Figure 2. Juniperus spp. Indian Perfumer 1980; 24:57–65. 10. Li N, Christian T, Farjon A, Rushforth K. Juniperus recurva var. coxii. The IUCN Red List of Threatened Species 2013: e.T34142A2847725, 2013. 11. 11.Weyaerstahl P, Marschal-Weyerstahl H, Manteuffel E, Kaul VK. Constituents of Juniperus recurva var. squamata oil. Planta Med 1988; 54(3):259–61. 12. Bhandari J, Naqvi A, Niraula P, Thapa P, Thapa N, Shrestha N, Shrestha BG. Phytochemical screening, antioxidant assay of Juniperus recurva and study of it’s in vitro cytotoxicity against breast cancer cell lines. Int J Pharm Bio Sci 2015; 6:1134–45. 13. Bais S, Prashar Y. Identification and characterization of

amentoflavone from six species of Juniperus against H2O2 induced oxidative damage in human erythrocytes and leucocytes. Res J Phytochem 2015; 2:1–15. 14. Kour A, Shawl AS, Rehman S, Sultan P, Qazi PH, Suden P, Khajuria RK, Verma V. Isolation and identification of an endophytic strain of Fusarium oxysporum producing podophyllotoxin from Juniperus recurva. World J Microbiol Biotechnol 2008; 24:1115–21. 15. Cowan M. Plant products as antimicrobial agents. Clin Microbiol Reviews 1990; 12:564–82. Fig. 2. Summary of Pharmacological Activities and Traditional Uses 16. Reische D, Lillard D, Eitenmiller R. Antioxidants in food lipids. In: of J. recurva Ahoh CC, Min DB (eds) Chemistry, Nutrition and Biotechnology, Marcel Dekker, New York, pp 423–48, 1998. 17. Buchbauer G. The detailed analysis of essential oils leads to the CONCLUSION understanding of their properties. Perfumer & Flavorist 2000; 25:64–7. In this review paper various traditional and medicinal 18. Bown D. The Royal Horticultural Society Encyclopedia of Herbs and aspects of J. recurva have been discussed. There is a shortage Their Uses. Dorling Kindersley Ltd, London, pp 424, 1999. 19. Watt O, Breyer-Brandwijk M. The medicinal and poisonus plants of research reports on J. recurva. It remains one of least of Southern and Eastern Africa. E & S Livingstone Ltd, Edinburgh & studied species of Juniperus. Further evaluation needs to be London, pp 841, 1962. carried out on J. recurva in order to establish and confirm 20. Clutton DM. History of Gin. Flavour. Ind, pp 454, 1972. 21. Takacsova M, Pribela A, Faktorova M, Nahrung. Study of the its medicinal uses and development of formulations antioxidative effects of thyme, sage, juniper and oregano. Nah rung containing this plant for their practical clinical applications, Food 1995; 39:241–3. 22. Zaman MB, Khan MS. Hundred drug plants of West Pakistan. which can be used for the welfare of mankind. Pakistan Forest Institute Peshawar, Pakistan, 1970. 23. Baquar SR. Medicinal and poisonous plants of Pakistan. Printas ACKNOWLEDGEMENTS Karachi, Pakistan, pp 506, 1989. 24. Oda J, Ando N, Nakajima Y, Inouye Y. Studies on insecticidal The authours are thankful to Dr. P. H. Qazi, Senior constituents of Juniperus recurva Buch Agr. Biol Chem 1977; 41:201–4. Scientist, Indian Institute of Integrative Medicine, Sanat 25. Wedge DE, Tabanca N, Sampson BJ, Werle C, Demirci B, Baser Nagar, Srinagar for providing laboratory support and KH, Nan P, Duan J, Liu Z. Antifungal and insecticidal activity of two valuable suggestions. We are thankful to reviewers for their Juniperus essential oils. Nat Prod Commun 2009; 4(1):123–7. 26. Mahajan J, Kumar R, Kaur R, Arora S. Exploration of xanthine oxidase inputs. producing microorganisms from hypoxanthine supplemented soil and their inhibitors from plants employing nitroblue tetrazolium based colorimetric assay method. Glo Adv Res J of Environ Sci CONFLICT OF INTERESTS Toxicol 2012; 1:10–3 27. Sati SC, Joshi S. Antibacterial potential of leaf extracts of Juniperus The authors declare that there is no conflict of interests communis L. from Kumaun Himalaya. Afr J Microbiol Res 2010; regarding the publication of this paper. 4:1291–4.

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28. Angioni A, Barra A, Russo AT, Coroneo A, Dessia S, Cabras P. 34. Stoilova IS, Wanner J, Jirovetz L, Trifonova D, Krastev L, Stoyanova Chemical composition of the essential oils of Juniperus from ripe AS, Krastanov AI. Chemical composition and antioxidant properties and unripe berries and leaves and their antimicrobial activity. J Agric of juniper berry (Juniperus communis L.) essential oil. Bulg J Agric Food Chem 2003; 5:3073–78. Sci 2014; 20:227–37 29. Pepeljnjak S, Kosalec I, Kalodera Z, Blazevic N. Antimicrobial 35. Saab AM, Guerrini A, Sacchetti G, Maietti S, Zeino M, Arend J, activity of juniper berry essential oil (Juniperus communis L., Gambari R, Bernardi F, Efferth T. Phytochemical analysis and Cupressaceae). Acta Pharm 2005; 55(4):417–22. cytotoxicity towards multidrug-resistant leukemia cells of essential 30. Filipowicz N, Kamiński M, Kurlenda J, Asztemborska M, Ochocka oils derived from Lebanese medicinal plants. Planta Med 2012; JR. Antibacterial and antifungal activity of juniper berry oil and its 78:1927–31. selected components. Phytother Res 2003; 17(3):227–31. 36. El-Sawi S, Motawae H, Ali M. Chemical composition, cytotoxic 31. Hoferl M, Stoilova I, Schmidt E, Wanner J, Jirovetz L, Trifonova activity and antimicrobial activity of essential oils of leaves and D, Krastev L, Krastonov A. Chemical composition and antioxidant berries of Juniperus phoenicea growing in Egypt. Afr J Trad Compl properties of Juniper Berry (J. communis L.) essential oil. Action Alt Med 2007; 4:417–26. of the essential oil on the antioxidant protection of Saccharomyces 37. Cabral C , Francisco V, Cavaleiro C, Gonçalves MJ, Cruz MT, Sales cerevisiae model organism. Antioxidants 2014; 3(1);81–98. F, Batista MT, Salgueiro L. Essential oil of Juniperus communis sub 32. Bukowska B. Effects of 2,4-D and its metabolite 2,4-dichlorophenol sp. alpina (Suter) Celak needles: chemical composition, antifungal on antioxidant enzymes and level of glutathione in human activity and cytotoxicity. Phytother Res 2012; 26(9):1352–7. erythrocytes. Comp Biochem Physiol C Toxicol Pharmacol 2003; 38. Uprety Y, Asselin H, Boon EK, Yadav S, Shrestha KK. Indigenous 135:435–41. use and bio-efficacy of medicinal plants in the Rasuwa District, 33. Cioanca O, Hancianu M, Mihasan M, Hritcu L. Anti- Central Nepal. J. Ethnobiol Ethnomed 2010; 6:3–9. acetylcholinesterase and antioxidant activities of inhaled Juniper oil on amyloid beta (1-42)-induced oxidative stress in the rat hippocampus. Neurochem Res 2015; 40(5):952–60.

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