A REVIEW ON SPIKENARD (NARDOSTACHYSJATAMANSI DC.)-AN ‘ENDANGERED’ ESSENTIAL HERB OF INDIA

The herb Nardostachys jatamansi DC. is known to be a popular medicinal and aromatic plant species. It is a reputed Ayurvedic herb and used in various multiple formulations. It has also been mentioned in the Holy Bible and Quran. The roots and rhizomes of Nardostachys jatamansi DC. have been used to treat epilepsy, hysteria, syncope and mental weakness.It also exhibits cardio protective activity and used in the treatment of neural diseases. The essential oil obtained from the roots of jatamansi showed various pharmacological activity including antimicrobial, antifungal, hypotensive, antiarrhythmic and anticonvulsant activity. Sesquiterpene is the major component of this plant, others include jatamansone, nardostachone. But this reputed plant species have become critically endangered and requires various conservation strategies. The review summarizes the conservation methodologies being investigated on the plant as well as its phytochemical and pharmacological investigations. Keyword: Nardostachys jatamansi, conservation, rhizome, India


Introduction
Nardostachys jatamansi DC. is a known medicinal herb which is on the verge of extinction. The dependence of the jatamansi species on habitats has mainly caused its exploitation, making the species critically endangered 1,2,3 , the rhizomes of the herb being over-exploited for medicinal use, habitat degradation and other biotic interferences. The earlier workers therefore gave emphasis on its conservation and multiplication 2,4,5,6,7 .Various methods should be used for its conservation, in situ as well as ex situ being a few. The rhizome of the herb is considered as the official plant part, found to have excellent therapeutic potential. As mentioned in Ayurveda, the roots and the rhizomes of Nardostachys jatamansi DC. are used to treat epilepsy, hysteria, syncope and mental weakness 8 .In India, the rhizomes and roots are being marketed as an anticonvulsant Ayurvedic drug known as Ayush 56 and also used as an antistress agent 9 .The rhizome of jatamansi is used as an aromatic adjunct in the preparation of medicinal oils, to promote hair growth and blackness 10 . The roots of the herb is used in the preparation of an essential oil found to have fungi toxic activity 11 , antimicrobial 12 , antifungal 13 , hypotensive 14 , antiarrhythmic 14 and anticonvulsant activity 14,15 .As the species is reported to have become critically endangered, effective conservation, management and recovery of the species is highly important. It can be deliberated through variability analysis. Study of reproductive biology is also necessary for effective conservation of endangered plants including study of pollination behaviour.

Historical background
The medicinal benefits of the plant has been valued for centuries in Indian Ayurveda, in Unani in ancient Greek and Arab, and in ancient Egypt and Rome. The N. jatamansi drug originates from 'Song Zhou' of Chuan Xi which has been registered in the book 'Ben Cao Shi Yi' in China. In the 'Book 18 of Homer's Iliad', the perfume of N. jatamansi is used Patroklos by Achilles. The herb has been mentioned twice in the biblical love poem, the 'Song of Solomon (1:12 and 4:13)'; it is one of the Eleven Herbs used for the Incense in the Holy Temple in Jerusalem. It has also been mentioned that Mary uses a pound of pure N. jatamansi to anoint Jesus's feet. The fruit which Adam ate in Paradise, which God had forbidden him to eat is believed to be the powdered root of N. jatamansi as mentioned in some Islamic traditions.In Medieval European cuisine, the jatamansi herb is used for seasoning the foods 16 .

Botany and taxonomy of spikenard
The sages of Ayurveda have broadly categorized the diseases according to adhishthana (place of origin) into two types: sharirik (physical) and manasik (mental). Among the various herbs mentioned in the texts as sangyasthapaka and manasdoshhar (psychotropic), jatamansi is the foremost of these herbs. It is known as sulomasa, bhutjata and tapaswini in Sanskrit and commonly known as spikenard, nard or muskroot. It belongs to the family Valerinaceae and its scientific classification is represented by table2. The Valeriana jatamansi which is similar to nardostachys is used as a substitute. Valerianaceae flowers and fruits are marked by a degree of differentiation in their morphology, characterized by sympetalous flowers, inferior 3 celled ovary, one fertile carpel, achene type of fruit and lack of endosperm 17 . Nardostachys jatamansi is an erect, perennial herb, reaching a height of 10-60 cm (Figure 1a) with woody rhizomatic rootstock ( Figure 1b) covered with tail like brown fibers, while the root penetrates deep in the soil. Leaves are long and narrow. Arising in terminal corymbose cymes, the flowers are creamy white, often rosy or pale pink in appearance. Fruit is small, 4 mm long, covered with minute hairs. It has been described as a combination of three tastes -bitter, astringent and sweet. Within the family Valerianaceae, the genus Nardostachys is considered as most primitive 17,18 . Table1 represents the scientific classification of Nardostachys jatamansi.

Occurrence and distribution
The jatamansi species is found to be distributed in the subalpine to alpine areas of India (Jammu and Kashmir, Himachal Pradesh, Uttarakhand, Sikkim), Pakistan, Nepal, Tibet, China and Yunan. The herb mostly grows between 3300-5000m above sea level. Due to over-exploitation of the rhizome, the species has become endangered in most of its natural populations 19,3 and critically endangered in few other populations 2 . Nautiyal et al. used vegetative propagules and seedling transplantation methods under different treatments to carry out cultivation trials of jatamansi at three altitudes (low -1800 m, middle -2200 m and natural habitat -3600 m). Variation in the economic yield occurred with treatment, age of plant at the time of transplantation and altitude of nursery.With the addition of manure (FYM) in all the treatments and altitudes, the economic yield increased as compared to control. Cultivation was not found commercially viable at 1800 m in any of the treatments due to total mortality. At 3600 m, both vegetatively grown as well as transplanted seedling crop showed marked profit in polyhouse condition 3 .

The Endangered species
The Himalayan region of India is known for its richness in medicinal plants with a total of 1,748 species. To safeguard economic and conservation concerns, the medicinal plants are attracting more attention 20  brown fibres forming a network which are skeletons of sheathing leaf bases), size (3 to 7 cm long, 0.5 to 1.5 cm in diameter), colour (dark brown externally and reddish brown internally), fracture (brittle), surface (fibrous), odour (strongly aromatic) and taste (Acrid, slightly bitter and aromatic). The physicochemical parameters were also studied in order identify various adulterants and improper handling of drugs. The physicochemical parameters were measured in % w/w giving the ash values as follows total ash (8.5), acid insoluble (3.8), water soluble (0.5) and sulphated ash ( Medicinal plants (MPs) and other non-timber forest products (NTFPs) are threatened by many factors, from both social and ecological perspectives. In response to differential harvesting strategies, the effect on demography of harvesting of the rhizome part of the plant varies greatly. The frequency of harvesting and the amount of jatamansi harvested should be strictly regulated, especially those growing in high altitude habitats on dry rocky outcrops. These rare and endangered Himalayan MPs require specific management plans based on their plant parts used, growth patterns in different habitats as well as the harvesting practices that are used 5 .

Phytochemistry
Ayurveda ascribes three characteristics to herbs: • Gunna (properties) -laghu (light) and snigdh (slimy) • Rasa (taste) -katu (pungent), madhur (sweet) and tickta (bitter) • Virya (potency) -sheet (cold) For any kind of drug discovery, prior knowledge of the chemical composition and their structure is utmost. Drug designing now depends more on rationality and evidences, thus evolving the discovery of drugs. Effective contributions have been made in fields such as cancer chemotherapy, drug resistant infections, and neurological diseases by structure-based drug designing. Developments in technology especially computers, bioassay techniques and calibrated instruments aid in discovering newer methods for drug designing as well as drug discovery. In modern medicine, computational structure-based drug designing is the key to finding novel treatments 31 . N. jatamansi has been discovered with both volatile and non-volatile constituents. Sesquiterpenes contribute to the major portion of the volatile compounds while sesquiterpenes, coumarins, lignans, neolignans, alkaloids and steroids form the major components of the nonvolatile extracts 32,33,34,35 . Sesquiterpene was eventually found in abundance in both the categories and found to accumulate in higher levels. Valeranone (jatamansone) is a sesquiterpene found to be present at up to 0.7%. Other sesquiterpenes include nardin, nardal, nardol, valerenal, jatamnsic acid, b-maline and patchouli alcohol 36 (Figure 2b). Besides these two, several coumarins were already known 9 . Sesquiterpenes and coumarins are present in considerable amount in the roots of jatamansi plant mainly responsible for its essential oil 9 . Figure 2c represents the principal sesquiterpene of this plant named jatamansone or valeranone 38,39 . Various other sesquiterpenes known are nardostachone (Figure 2d), dihydro jatamansin, jatamansinol, jatamansic acid 40 , jatamansinone, jatamansinol, oroseolol, oroselone, seselin, valeranal, nardostachyin5, nardosinone, spirojatamol 41 , jatamol A and B 42 , calarenol 43 , seychellene, seychelane, coumarin: jatamansin or xanthogalin 44,45 . An alkaloid named actinidine has also been reported (Figure 2e). Nardal has been reported to be an active component by Venkateshwara (Figure 2f

Jatamansone (valeranone):
Antioestrogenic, anti-arrhythmic, antihypertensive, anticonvulsant, sedative and tranquilizing activities have been reported of jatamansone through animal studies 47,48,49,50 . Jatamansone does not have neuroleptic characteristics, still exhibits anticonvulsive effect against electric shock and gastro protective actions. Toxicological investigation on rats and mice were carried out showing an oral LD 50 greater than 3160 mg/kg, thus suggesting the chances of developing a therapeutically useful dose ratio 38 . Additionally, tranquillizer experiments exhibit jatamansone ability to prolong barbiturate hypnosis, impair rota rod performance and potentiate the bodytemperature lowering activity of reserpine. Pharmacological effects of jatamansone also includes in its activity towards reduction in aggressiveness, restlessness, stubbornness and insomnia. Jatamasnone, D-amphetamine and chlorpromazine were compared in a study which was conducted on hyperkinetic children for its efficacy. The behavior of these children was significantly improved by jatamasnone and amphetamine with amphetamine reducing aggressiveness and restlessness to a greater extent. Jatamansone shows less side effects as compared to D-amphetamine and chlorpromazine 51. 6.2 Jatamansin: Jatamansin is a coumarin known to be effective in internal treatment of varicose veins 52 . Ethanolic extracts of N. jatamansi (NJE) were administered in Wister rats in two doses followed by immobilization stress on the 5 th day.Rats that received different doses of NJE did not show clinical signs of toxicity and in vitro study showed free radical scavenging activity of NJE, as evidenced by low IC 50 value. Pre-treatment with NJE-200 and 500 mg/kg significantly decreased lipid peroxidation (LPO) and nitrite level and increased the catalase activity in the brain. Presence of flavonoids and polyphenols is responsible for the antioxidant property of NJE and may also be responsible for its anti-stress effect 54 . The antioxidant and anti-cataleptic effects of the aqueous extract of Nardostachys jatamansi roots was studied. Male albino rats were administered with haloperidol (1 mg/kg, ip) to induce catalepsy. Though a significant (P < 0.01) reduction was observed in the cataleptic scores of all the drug-treated groups as compared to the haloperidol-treated group, maximum reduction was observed in the group treated with Nardostachys jatamansi (250 and 500 mg/kg body weight). Among the biochemical parameters, generation of thio-barbituric acid reactive substances (TBARS) increased while glutathione (GSH) reduced significantly as a result of haloperidol administration. However, after their treatment with Nardostachys jatamansi, the substances were restored to near normal level. Haloperidol administration reduced the levels of catalase and superoxide dismutase (SOD) significantly which were restored to their normal level by Nardostachys jatamansi treatment. Thus Nardostachys jatamansi reverses the haloperidol-induced catalepsy in rats and reduces oxidative stress 55  jatamansi. The study showed major hepatoprotective activity against thioacetamide induced hepato-toxicity and reduced the levels of serum transaminase andalkaline phosphatase significantly. An increase in the survival rates in rats intoxicated with LD 90 dose of the hepatotoxic drug was highly observable after pretreatment of the animals with the extract 58  The right striatum was infused with 2 µl of 6-OHDA (12 µg in 0.01% in ascorbic acid-saline) on day 21, while 2 µl of vehicle was infused in the sham-operated group. The neuro behavioural activity in the rats were tested after three weeks of 6-OHDA injection which were then sacrificed after 6 weeks in order to evaluate lipidperoxidation, reduced glutathione content, the activities of glutathione-S-transferase, glutathione reductase, glutathione peroxidase, superoxide dismutase and catalase, quantification of catecholamines, dopaminergic D2 receptor binding and tyrosinehydroxylase expression. The alterations caused by 6-OHDA injections such as increase in drug-induced rotations and deficits in locomotor activity and muscular coordination were extensively and dose-dependently restored by N. jatamansi. Thus attenuation of Parkinsonism might be enhanced using the extractof N. jatamansi as indicated by this study 63 . 7.9 Anti-alzheimer's activity: A strong linkage between Insomnia and Alzheimer's disease (AD) has been hypothesized. With age, sleep disturbances and dementia increases as per statistical data and chances of developing AD and depression in young and middle-aged adults suffering from insomnia is 11 times more likely in their later life. The effect of methanolic extract of Nardostachys jatamansi DC. (MENJ) rhizome was investigated on sleep deprived (SD) amnesic mice in order to assess anti-amnesic activity. Pretreatment of animals with MENJ and Piracetamas standard drug for 14 days was followed by 5 days sleep deprivation using multiple platform method.Groups that were treated with MENJ doses showedconsiderable improvement in learning and cognition parameters in behavioral tests. The loss of memory and cognitive deficits due to sleep deprivation could be protected by MENJ as suggested by the above study 64 . Rahman et al. investigated the anti-amnesic activity of methanolic extract of rhizome of Nardostachys jatamansi DC. (MENJ) on sleep deprived (SD) amnesic mice. Pretreatment of animals with MENJ and Piracetam for 14 days was followed by sleep deprivation using multiple platform method for 5 days. The mice brain was