Bhutanese Traditional Medicine Research Articles

Strengthening Bhutanese Traditional Medicine in healthcare service delivery in Bhutan

Bhutanese Traditional Medicine, known as Sowa Rigpa, deeply rooted in the country’s cultural and religious heritage, plays a significant role in healthcare service delivery in Bhutan. This commentary explores the evolution and current landscape of Sowa Rigpa within the context of the nation’s healthcare system. While traditional medicine services have expanded over the years, challenges such as scepticism regarding scientific validity, inadequate infection control measures and concerns about the sustainability of medicinal plants persist. To address these challenges, recommendations, including the integration of modern technology in quality assurance, comprehensive training in infection control, evidence-based practice and conservation efforts for medicinal plants have been adopted. By embracing these strategies, Sowa Rigpa can enhance its efficacy, safety and sustainability, ensuring its continued relevance in meeting the healthcare needs of the Bhutanese population while contributing to the global advancement of Traditional Medicine practices.

The Diversity of Medicinal Plants of the Himalayan Kingdom of Bhutan

Most nations depend heavily on plants as economic resources. They can be categorized according to their contribution to economic development thanks to their taxonomic classes. Plant components are used in pharmaceutical research and the creation of novel medications in the healthcare industry. Plant components or herbal concoctions are utilized as herbal medicines in Bhutan and other parts of the world to treat various illnesses. The study aimed to list all the plants possessing medicinal properties from the published sources. The proponents of herbal medicine in Bhutan assert that plant parts contain a variety of phytochemicals that show a range of pharmacological and biological responses. The related data were searched online using various boolean operations. Bhutan is home to around 637 medicinal plant species under 132 families and 418 genera, of which only 229 species are used in Bhutanese traditional medicine. More research and interactions with the local traditional medicinal practitioners can reveal more plants with medicinal properties.

Are people aware of the Buddhist astrological recommendations on timing for surgery?

Introduction: Adhering to the astrological recommendations on timing of invasive therapies are a standard practice in gSo-wa Rig-pa based Bhutanese Traditional Medicine. The day (for La-dza, Sog-dza and Shey-dza), lunar date (for La-ney) and time within the day of surgery (for La-ney), along with Wang-thang (field of power) and Lung-ta (Horse of fortune) are important astrological recommendations. However, there are no information about the patients’ knowledge, attitude and practice on astrological recommendations for timing of surgery in an allopathic setting in Bhutan. The study was conducted to assess the knowledge, attitude and practice of considering Buddhist astrological recommendations among patients undergoing elective surgeries. Methods: A cross-sectional study with convenient sampling was conducted at the National Referral Hospital, Thimphu from April to May 2018. Data were collected using a pretested interviewer-administered questionnaire. Ethical clearance was obtained from the Research Ethics Board of Health, Bhutan. Results: From 420 participants (response rate 99.5%), 188 (44.8%) were aware that there are astrologically determined good timings to undergo elective surgeries. Only four persons (1.0%) were aware about La-ney. Among those who believed in such astrological recommendations, the attitude were more favourable towards the areas in which they had higher knowledge. Conclusion: The patients had very poor knowledge about the astrological recommendations to choose the timing for surgery. A well designed study on the effect of astrological recommendations on the surgical outcomes is recommended for generating evidence for further action.

Patients’ knowledge and practices on the drug regimens prescribed at the National Traditional Medicine Hospital: a call for a systematic approach to drug information services

Introduction: The World Health Organisation estimates that only half of all medicines dispensed are used appropriately. The Bhutanese Traditional Medicine (BTM) system is no exception. Objectives: We studied the patients’ knowledge on the usage of traditional medicine regimens prescribed to them, their medication adherence and possible adverse events.
 Methods: This was a cross-sectional study conducted using an interviewer-administered questionnaire at the National Traditional Medicine Hospital, Thimphu in 2017. A simple random sampling was used. Ethical approval was sought from Research Ethics Board of Health, Thimphu.
 Results: A sample of 402 patients were studied. The mean age of the respondents was 45 years (±15 years) and 333 (82.8%) were old cases presenting to the hospital. The knowledge on the correct timing of the medicine, how to take the medicine and what to avoid while on those medicines were understood only by 62.7%, 34.6% and 22.9% respectively. Only 351 (87.3%) patients received verbal information on the usage of drugs from the dispensers, of which 119 (29.6%) said the information was not clear or not adequate. Failure to take medicines on time was reported by 224 (67.3%) among old cases. Undesirable events reported while using BTM were giddiness, vomiting, skin irritation, diarrhoea and fainting episodes.
 Conclusions: The drug information system and monitoring of medication adherence and adverse drug events require adoption of more effective toolsand methods.

Geopharmaceuticals of Himalayan Sowa Rigpa medicine: Ethnopharmacological uses, mineral diversity, chemical identification and current utilization in Bhutan

Ethnopharmacological relevanceGeological materials, such as minerals, have a long history of usage as ingredients in multicompound formulations of Himalayan Sowa Rigpa medicine – as well as in its localized form of Bhutanese traditional medicine (BTM) – for treating various disorders for over thousand years. Yet, hardly any scientific research has been done on their ethnopharmacological efficacy and chemistry. Aim of the studyThis study documents and correlates the rarely explored ethnopharmacological and chemical identification of various minerals and their ethnomedicinal uses in BTM formulations for the first time. Material and methodsA five stage cross-disciplinary process was conducted as follows: (1) a review of classical literature of Sowa Rigpa texts (Tibetan medical texts, pharmacopoeias and formularies) that are still in use today; (2) listing of mineral ingredients according to Sowa Rigpa names, followed by identification with common English and chemical names, as well as re-translating their ethnomedical uses; (3) cross-checking the chemical names and chemical composition of identified Sowa Rigpa minerals with various geological mineral databases and mineral handbooks; (4) authentication and standardization of Sowa Rigpa names through open forum discussion with diverse BTM practitioners; (5) further confirmation of the chemical names of identified minerals by consulting different experts and pharmacognosists. ResultsOur current study lists 120 minerals as described in Sowa Rigpa medical textbooks most of which we were able to chemically identify, and of which 28 are currently used in BTM herbo-mineral formulations. Out of these 28 mineral ingredients, 5 originate from precious metal and stone, 10 stem from earth, mud and rocks, 8 are salts, and 5 concern ‘essences’ and exudates. ConclusionsOur study identified 120 mineral ingredients described in Sowa Rigpa medical textbooks, out of which 28 are currently used. They are crucial in formulating 108 multicompound prescription medicines in BTM presently in use for treating more than 135 biomedically defined ailments.

A medicinal plant compound, capnoidine, prevents the onset of inflammation in a mouse model of colitis

Ethnopharmacological relevanceThe traditional uses of Corydalis dubia, Ajania nubigena and Pleurospermum amabile in the Bhutanese traditional medicine for treating disorders related to inflammatory conditions and the in vitro anti-inflammatory activity of their crude extracts inspired the isolation and the investigation of anticolitic properties of four pure compounds. Materials and methodsThree medicinal plants were collected from Himalayan Mountains of Bhutan. Capnoidine and scoulerine were isolated from C. dubia, linalool oxide acetate from A. nubigena and isomyristicin from P. amabile using natural product isolation protocols. Four compounds were investigated for their anti-inflammatory activities against IBD-colitis using chemically induced (TNBS) mice model of colitis. Capnoidine conferred the best preliminary protection against TNBS-induced colitis in mice and we have conducted in-depth pharmacological investigation of this compound including clinical symptoms, pathological signs, cytokine profiles, histological structure and inflammasomes using relevant bioassay protocols. ResultsCapnoidine-treated mice had significantly: a) improved clinical symptoms (body weight loss, mobility, piloerection and faecal consistency); b) reduced colon pathology (adhesion, oedema, ulceration, and colon length); c) altered inflammatory cytokines profiles within the colons; d) reduced levels of p-IκB-α (Ser32) and p-NF-κB p65 (Ser536) and e) reduced histological inflammation in the colon when compared with mice administered TNBS only. ConclusionCapnoidine presents as a potential new anti-inflammatory drug lead candidate for diseases where current standard-of-care often fails and is associated with major side effects. It also validates the traditional uses of C. dubia against inflammatory conditions and underlines the value of pursuing bioactive compounds derived from traditionally used ethnobotanical medicines.

An approach to Illnessess by Traditional Medicine of Bhutan: An Introduction

Depending on the different ailments, various diagnostic tools like laboratory tests, X-ray, ultrasound, magnetic resonance imagery, and computerized topography scan are used in modern medicine. However, in Bhutanese traditional medicine the method of diagnosis is based on the medical text of Sowa-Rigpa, the ancient practices involving physical examination, questioning, pulsation, palpation and analysis of excretion. These enduring techniques of diagnosis use no modern technologies. The system has been practised in Bhutan for hundreds of years and has been officially establishment in 1967. Insights of this article are compiled from the Buddhist medical text and oral transmission of senior physicians. We discuss diagnosis in three categories namely, visual (Ta-Wa), touch (Reg-Pa) and questioning (Dri-Wa). Visual diagnosis mainly investigates the urine, physical appearance and the tongue. Touch, reading the pulse through which the physician diagnose illnesses. Questioning about the signs and symptoms of the illnesses and past medical history.

Compounds Derived from the Bhutanese Daisy, Ajania nubigena, Demonstrate Dual Anthelmintic Activity against Schistosoma mansoni and Trichuris muris.

BackgroundWhipworms and blood flukes combined infect almost one billion people in developing countries. Only a handful of anthelmintic drugs are currently available to treat these infections effectively; there is therefore an urgent need for new generations of anthelmintic compounds. Medicinal plants have presented as a viable source of new parasiticides. Ajania nubigena, the Bhutanese daisy, has been used in Bhutanese traditional medicine for treating various diseases and our previous studies revealed that small molecules from this plant have antimalarial properties. Encouraged by these findings, we screened four major compounds isolated from A. nubigena for their anthelmintic properties.Methodology/Principal FindingsHere we studied four major compounds derived from A. nubigena for their anthelmintic properties against the nematode whipworm Trichuris muris and the platyhelminth blood fluke Schistosoma mansoni using the xWORM assay technique. Of four compounds tested, two compounds—luteolin (3) and (3R,6R)-linalool oxide acetate (1)—showed dual anthelmintic activity against S. mansoni (IC50 range = 5.8–36.9 μg/mL) and T. muris (IC50 range = 9.7–20.4 μg/mL). Using scanning electron microscopy, we determined luteolin as the most efficacious compound against both parasites and additionally was found effective against the schistosomula, the infective stage of S. mansoni (IC50 = 13.3 μg/mL). Luteolin induced tegumental damage to S. mansoni and affected the cuticle, bacillary bands and bacillary glands of T. muris. Our in vivo assessment of luteolin (3) against T. muris infection at a single oral dosing of 100 mg/kg, despite being significantly (27.6%) better than the untreated control group, was markedly weaker than mebendazole (93.1%) in reducing the worm burden in mice.Conclusions/SignificanceAmong the four compounds tested, luteolin demonstrated the best broad-spectrum activity against two different helminths—T. muris and S. mansoni—and was effective against juvenile schistosomes, the stage that is refractory to the current gold standard drug, praziquantel. Medicinal chemistry optimisation including cytotoxicity analysis, analogue development and structure-activity relationship studies are warranted and could lead to the identification of more potent chemical entities for the control of parasitic helminths of humans and animals.

Diterpenoid alkaloids of Aconitum laciniatum and mitigation of inflammation by 14-O-acetylneoline in a murine model of ulcerative colitis.

Aconitum laciniatum is used in Bhutanese traditional medicine for treating various chronic infections and inflammatory conditions. We carried out in-depth isolation and characterization of the phytochemicals from the root component and determined the anti-inflammatory effects of the isolated compounds against chemically-induced colitis in mice. Five diterpenoid alkaloids - pseudaconitine, 14-veratroylpseudaconine, 14-O-acetylneoline, neoline, and senbusine A - were isolated from A. laciniatum for the first time. Two of the alkaloids were tested for anti-inflammatory properties in the TNBS-induced colitis model in mice. Various parameters were measured to assess pathology including weight loss, clinical and macroscopic scores, histological structure and IFN-γ production in the gut. Of the two alkaloids tested, 14-O-acetylneoline showed significant protection against different parameters of colitic inflammation. Compared to control mice that received TNBS alone, mice treated with 14-O-acetylneoline experienced significantly less weight loss and had significantly lower clinical scores, macroscopic pathology and grades of histological inflammation. Moreover, colonic IFN-γ mRNA levels were significantly reduced in mice that received 14-O-acetylneoline compared to control mice that received TNBS alone. This alkaloid is now considered a novel anti-colitis drug lead compound.

Contributions of medicinal plants to the Gross National Happiness and Biodiscovery in Bhutan.

BackgroundThe medicinal plants and the associated Bhutanese traditional medicine (BTM) are protected by the country’s constitution and receive both government support and acceptance by the wider public. More than 1000 medicinal plants are described in the BTM but currently collects only 300 species for daily formulations of BTM. These medicinal plants have been one of the drivers of the ‘Gross National Happiness (GNH)’ and biodiscovery projects in Bhutan. However, no review covering the systematic evaluations of the contributions of medicinal plants and the BTM to the GNH and biodiscovery exist till date.MethodsThis paper, therefore addresses this information gap. It is based on the review of the existing traditional and scientific literature, government websites and policy documents. The descriptions and discussions of the paper is straightened, authenticated and enhanced by the data collected through the informal discussions with the BTM practitioners and also through the authors’ many years of practical observations of the impact of the medicinal plants programs and the BTM practices in Bhutan.ResultsThis paper found the following: a) the medicinal plants generates income to the farmers elevating their living standard and the economic status, b) it serves as the bulk ingredients of the BTM facilitating the provision of free traditional health care services to the patients, c) helps the conservation of medicinal plants and their pristine environment through recognition of their spiritual, social and economic values, d) preserves the rich BTM cultural heritage, and e) guides the biodiscovery projects based on their ethnobotanical information. The paper also identified the challenges and research gaps, and recommends appropriate strategies that can help secure the sustainable future of the medicinal plants, the BTM and the biodiscovery projects.ConclusionsThe medicinal plants play significant role in the country’s biodiscovery projects and the internationally renowned development policy of ‘Gross National Happiness’.

A new protoberberine alkaloid from Meconopsis simplicifolia (D. Don) Walpers with potent antimalarial activity against a multidrug resistant Plasmodium falciparum strain

Ethnopharmacological relevanceThe aerial components of Meconopsis simplicifolia (D. Don) Walpers are indicated in Bhutanese traditional medicine for treating malaria, coughs and colds, and the infections of the liver, lung and blood. This study is to validate the ethnopharmacological uses of this plant and also identify potent antimalarial drug leads through bioassays of its crude extracts and phytochemical constituents. Materials and methodsMeconopsis simplicifolia (D. Don) Walpers was collected from Bhutan and its crude MeOH extract was subjected to acid-base fractionation. Through repeated extractions, separations and spectroscopic analysis, the alkaloids obtained were identified and tested for their antimalarial and cytotoxicity activities. ResultsPhytochemical studies resulted in the isolation of one new protoberberine type alkaloid which we named as simplicifolianine and five known alkaloids: protopine, norsanguinarine, dihydrosanguinarine, 6-methoxydihydrosanguinarine and oxysanguinarine. Among the five of the alkaloids tested, simplicifolianine showed the most potent antiplasmodial activities against the Plasmodium falciparum strains, TM4/8.2 (chloroquine–antifolate sensitive strain) and K1CB1 (multidrug resistant strain) with IC50 values of 0.78μg/mL and 1.29μg/mL, respectively. The compounds tested did not show any significant cytotoxicity activities against human oral carcinoma KB cells and normal Vero cells of African kidney epithelial cells. ConclusionsThis study validated the traditional uses of the plant for the treatment of malaria and identified a new alkaloid, simplicifolianine as a potential antimalarial drug lead.

Inhibition of TNF-α production in LPS-activated THP-1 monocytic cells by the crude extracts of seven Bhutanese medicinal plants

Ethnopharmacological relevanceSeven studied medicinal plants; Aconitum laciniatum, Ajania nubigena, Codonopsis bhutanica, Corydalis crispa, Corydalis dubia, Meconopsis simplicifolia and Pleurospermum amabile, are currently used in the Bhutanese Traditional Medicine (BTM) for the management of different types of disorders including the diseases that bore relevance to various inflammatory conditions. Aims of the studyThis study aimed to evaluate the inhibition of TNF-α production in LPS-activated THP-1 monocytic cells by the crude extracts of seven selected Bhutanese medicinal plants. It is expected to; (a) generate a scientific basis for their use in the BTM and (b) form a basis for prioritization of the seven plants for further phytochemical and anti-inflammatory studies. Materials and methodsSeven plants were selected using an ethno-directed bio-rational approach and their crude extracts were prepared using four different solvents (methanol, hexane, dichloromethane and chloroform). The TNF-α inhibitory activity of these extracts was determined by cytokine-specific sandwich quantitative enzyme-linked immunosorbent assays (ELISAs). The results were quantified statistically and the statistical significance were evaluated by GraphPad Prism version 5.01 using Student's t-test with one-tailed distribution. A p-value ≤0.05 was considered statistically significant. ResultsOf the seven plants studied, the crude extracts of six of them inhibited the production of pro-inflammatory cytokine, TNF-α in LPS-activated THP-1 monocytic cells. Amongst the six plants, Corydalis crispa gave the best inhibitory activity followed by Pleurospermum amabile, Ajania nubigena, Corydalis dubia, Meconopsis simplicifolia and Codonopsis bhutanica. Of the 13 extracts that exhibited statistically significant TNF-α inhibitory activity (p<0.05; p<0.01), five of them showed very strong inhibition when compared to the DMSO control and RPMI media. ConclusionsSix medicinal plants studied here showed promising TNF-α inhibitory activity. These findings rationalize the traditional use of these selected medicinal plants in the BTM as an individual plant or in combination with other ingredients for the treatment of disorders bearing relevance to the inflammatory conditions. The results forms a good preliminary basis for the prioritization of candidate plant species for an in-depth phytochemical study and anti-inflammatory activity screening of the pure compounds contained within those seven plants.

An assessment of the Bhutanese traditional medicine for its ethnopharmacology, ethnobotany and ethnoquality: Textual understanding and the current practices

Ethnopharmacological relevance: This study involves the assessment of the Bhutanese traditional medicine (BTM) which was integrated with the mainstream biomedicine in 1967 to provide primary health care services in the country. It caters to 20–30% of the daily out-patients within 49 traditional medicine units attached to 20 district modern hospitals and 29 Basic Health Units in the country. Aim of the study: This study presents the ethnopharmacological, ethnobotanical and the ethnoquality concepts in relation to mainstream Tibetan medicine and describes the current practices of BTM. Materials and methods: Experienced BTM practitioners (Drung-tshos and Smen-pas) were selected using a convenience sampling method and were interviewed using an open questionnaire followed by informal discussions. The corpus of BTM, Tibetan and scientific literature was obtained and the information on ethnopharmacological, ethnoquality and ethnobotanical concepts and current practices of BTM was extracted. Results: This study found that the BTM shares many similarities in terms of materia medica, pharmacopoeia and the principles and concepts of ethnopharmacology and ethnobotany with its mainstream Tibetan medicine. However, the resourceful Bhutanese Drung-tshos and Smen-pas have adapted this medical system based on the local language, culture, disease trend, health care needs and their familiarity with the locally available medicinal ingredients making it particular to the country. A number of notable distinctions observed in the current practices include a code of classification of diseases (only 79 of 404 types of disorders recognized), formulations (currently used only 103 of thousands formulation types), usage of medicinal plants (only 229 species of thousands described) and selected treatment procedures (golden needle and water therapy). This BTM was found to cater to 20–30% of daily out-patients visiting 49 modern hospitals and basic health units in the country. Conclusions: The BTM has been evolved from the Tibetan medicine. While the pharmacopoeia, ethnopharmacology, ethnobotany and the ethnoquality aspects shares commonalities with the mainstream Tibetan medicine, there are some practices unique to BTM. Such uniqueness observed in the current practices of BTM include formulations, medicinal plants collection and usage, and the treatment procedures including golden needle and water therapy. This could be a promising source of information for the rediscovery of useful remedies, the development of modern phytotherapeutics and the establishment of efficient quality control measures.

Antimalarial alkaloids from a Bhutanese traditional medicinal plant Corydalis dubia

Ethnopharmacological relevanceCorydalis dubia is used in Bhutanese traditional medicine as a febrifuge and for treating infections in the blood, liver and bile which correlate to the signs and symptoms of malarial and microbial infections. Aim of the studyTo validate the ethnopharmacological uses of the plant and to discover potential new therapeutic drug leads. Materials and methodsC. dubia was collected from Bhutan and the alkaloids were obtained using acid–base fractionation and separation by repeated column and preparative plate chromatography. The alkaloids were identified from analysis of their physiochemical and spectroscopic data and were tested for antiplasmodial, antimicrobial and cytotoxicity activities. ResultsA systematic extraction and isolation protocol yielded one new natural product, dubiamine, and seven known isoquinoline alkaloids, scoulerine, cheilanthifoline, protopine, capnoidine, bicuculline, corydecumbine and hydrastine. Among the four alkaloids tested, scoulerine showed the best antiplasmodial activity with IC50 values of 5.4μM and 3.1μM against the antifolate sensitive and the multidrug resistant P. falciparum strains: TM4/8.2 and K1CB1, respectively. None of the alkaloids tested showed significant antimicrobial or cytotoxicity activities. ConclusionsThe antiplasmodial test results, of the isolated alkaloid components, are commensurated with the ethnopharmacological uses of this plant.

Phytochemical and Biological Activity Studies of the Bhutanese Medicinal Plant Corydalis crispa

The chemical constituents and biological activities of Corydalis crispa (Fumariaceae) were investigated for the first time. The phytochemical study resulted in the isolation of nine known isoquinoline alkaloids: protopine (1), 13-oxoprotopine (2), 13-oxocryptopine (3), stylopine (4), coreximine (5), rheagenine (6), ochrobirine (7), sibiricine (8) and bicuculline (9), with complete NMR data for 2 and 3 provided here for the first time. Crude extracts exhibitedsignificant anti-inflammatory (p < 0.01) activity against TNF-alpha production in LPS activated THP-1 cells. The acetylcholinesterase inhibitory activity of compounds 2, 4 and 7 and the antiplasmodial activity of compound 5 against P. falciparum strains TM4/8.2 and K1CB1 (multidrug resistant strain) are reported here for the first time. Stylopine (4) did not show antimalarial activity against the K1CB1 strain in contrast to a previous report. This study generated a scientific basis for the use of this plant in Bhutanese traditional medicine, either individually or in combination with other medicinal ingredients to treat a broad range of disorders. This study also identified compound 5 as potential new antimalarial lead compound.

Evaluation of an ethnopharmacologically selected Bhutanese medicinal plants for their major classes of Phytochemicals and biological activities

As many as 229 medicinal plants have been currently used in the Bhutanese Traditional Medicine (BTM) as a chief ingredient of polyherbal formulations and these plants have been individually indicated for treating various types of infections including malaria, tumor, and microbial. We have focused our study only on seven species of these plants. We aim to evaluate the antiplasmodial, antimicrobial, anti-Trypanosoma brucei rhodesiense and cytotoxicity activities of the seven medicinal plants of Bhutan selected using an ethno-directed bio-rational approach. This study creates a scientific basis for their use in the BTM and gives foundation for further phytochemical and biological evaluations which can result in the discovery of new drug lead compounds. A three stage process was conducted which consisted of: (1) an assessment of a pharmacopoeia and a formulary book of the BTM for their mode of plant uses; (2) selecting 25 anti-infective medicinal plants based on the five established criteria, collecting them, and screening for their major classes of phytochemicals using appropriate test protocols; and (3) finally analyzing the crude extracts of the seven medicinal plants, using the standard test protocols, for their antiplasmodial, antimicrobial, anti-Trypanosoma brucei rhodesiense and cytotoxicity activities as directed by the ethnopharmacological uses of each plant. Out of 25 medicinal plants screened for their major classes of phytochemicals, the majority contained tannins, alkaloids and flavonoids. Out of the seven plant species investigated for their biological activities, all seven of them exhibited mild antimicrobial properties, five plants gave significant in vitro antiplasmodial activities, two plants gave moderate anti-Trypanosoma brucei rhodesiense activity, and one plant showed mild cytotoxicity. Meconopsis simplicifolia showed the highest antiplasmodial activity with IC(50) values of 0.40 μg/ml against TM4/8.2 strain (a wild type chloroquine and antifolate sensitive strain) and 6.39 μg/ml against K1CB1 (multidrug resistant strain) strain. Significantly the extracts from this plant did not show any cytotoxicity. These findings provide the scientific basis for the use of seven medicinal plants in the BTM for the treatment of malaria, microbial infections, infectious fevers, and the Trypanosoma brucei rhodesiense infection. The results also form a good preliminary basis for the prioritization of candidate plant species for further in-depth phytochemical and pharmacological investigations toward our quest to unearth lead antiparasitic, anticancer and antimicrobial compounds.

Antiplasmodial activity of atisinium chloride from the Bhutanese medicinal plant, Aconitum orochryseum

Ethnopharmacological relevance The plant Aconitum orochryseum Stapf. (Ranunculaceae) is employed together with other plants in Bhutanese traditional medicine and is indicated for malaria-associated fever. Aim of the study To study the in vitro antiplasmodial activity of atisinium chloride, the major alkaloid from Aconitum orochryseum. Materials and methods Atisinium chloride was extracted and purified from aerial parts of Aconitum orochryseum and its structure and absolute configuration confirmed by single crystal X-ray crystallography. The crude methanol extract, crude alkaloid fraction, and atisinium chloride were tested for in vitro antiplasmodial activity against the malarial Plasmodium falciparum strains TM4/8.2 (TM4; wild type) and K1CB1 (K1; chloroquine and antifolate resistant). Results The diterpenoid alkaloid atisinium chloride was shown to have moderate antiplasmodial activities with IC 50 values of 4 μM and 3.6 μM, respectively against the TM4 strain and the K1 strain of Plasmodium falciparum. Conclusions Our studies provide the first evidence in support of one of the indicated treatments with Aconitum orochryseum in Bhutanese traditional medicine. This alkaloid also represents a potential new antimalarial structural lead.

Risk Factors for the Sustainability of Medicinal Plants in Bhutan

AbstractMedicinal plants are used worldwide in primary health care and also for modern drug discovery programs. More than 13,000 species of Medicinal and Aromatic Plants are used in traditional medicines and herbal cosmetics throughout the world and about 8000 of these are used in South Asia alone. In Bhutan, while the total number of species used by Local Healing System(s) is unknown, more than 500 species of medicinal plants are recorded in the pharmacopoeia of Bhutanese traditional medicine called g.so-ba-rig-pa (Sowa Rigpa). Currently 300 species, which grow in diverse ecological zones of the country, are used by the Institute of Traditional Medicine Services in formulating 98 different essential Sowa Rigpa medicines. In 2007, 16 tons of medicinal plants were procured by the institute alone and 85% of them, almost all wild species, were collected within the country. Unless properly managed, the sustainability of these wild medicinal plants is likely to be threatened. In this light, our paper describes the possible risk factors that may affect the sustainable use of medicinal plants in Bhutan. Four broad risk factors, which include biological, ecological, social and economic issues, are identified here. Our paper also throws light to the current policies, frameworks and acts that are put in place to ensure the sustainable use of medicinal plants in Bhutan and finally suggests future directions.

Antiplasmodial agents from the Bhutanese medicinal plant Corydalis calliantha

The alkaloidal components of the Bhutanese medicinal plant Corydalis calliantha Long, which is used for the treatment of malaria, have been assessed. Four known alkaloids, protopine (1), scoulerine (2), cheilanthifoline (3) and stylopine (4) are reported from this plant for the first time. The protopine alkaloid, protopine, and the tetrahydroprotoberine alkaloid, cheilanthifoline, showed promising in vitro antiplasmodial activities against Plasmodium falciparum, both wild type (TM4) and multidrug resistant (K1) strains with IC(50) values in the range of 2.78-4.29 microM. Such activity had not been demonstrated previously for cheilanthifoline. The results thus support, at a molecular level, the clinical use of this plant in the Bhutanese traditional medicine and identified cheilanthifoline as a potential new antimalarial drug lead.

Hetisine-Type Diterpenoid Alkaloids from the Bhutanese Medicinal Plant Aconitum orochryseum

Investigation of Aconitum orochryseum, a Bhutanese traditional medicine ("gSo-ba Rig-pa") plant locally known as "Bong-kar", resulted in the isolation of three new hetisine-type diterpenoid alkaloids, named orochrine (1), 2-O-acetylorochrine (2), and 2-O-acetyl-7alpha-hydroxyorochrine (3), together with the previously reported alkaloids atisinium chloride (4) and virescenine (5). The structures of the new alkaloids were elucidated by spectroscopic data analysis.