Aconitum Species Research Articles

Morpho-phenological and Antibacterial Characteristics of Aconitum spp.

Aconitum species have been traditionally used as ethnomedicine to cure various ailments. The present study reveals the morpho-phenology and antibacterial property of alkaloid extracts of the two Aconitum species. The morpho-phenological characteristics will be helpful for determining the resource availability. Aconitum nagarum is erect type, whereas, Aconitum elwesii is a climber. Aconitum elwesii grows in advance of A. nagarum in terms of growth, flowering and senescence. Towards the end of the year, when the fruits have ripened, the parent tuber dies off. As a result, the daughter tuber becomes independent and in the following spring, takes over the function of the parent tuber. Aconitum nagarum and A. elwesii were found to contain 4-5 aconitine equivalent (AE) mg/g of alkaloid. These alkaloids showed antibacterial activity against different bacterial species including human pathogens, namely, Staphylococcus aureus, Salmonella typhimurium, Bordetella bronchiseptica, Escherichia coli, Bacillus subtilis, Pseudomonas putida, Pseudomonas fluorescence and Xanthomonas campestris. However, the extent of antibacterial activity varied among different bacterial species. The antibacterial activity against S. aureus, B. bronchiseptica, and B. subtilis was bactericidal in nature, whereas, against other tested bacterial species was bacteriostatic. Efficacy of the antibacterial activity of these alkaloids was evaluated by comparing with that of standard antibiotics. Differential localization of the antibacterial principle was observed among the Aconitum species studied.

ABSORPTION OF HYPACONITINE AND P-GLYCOPROTEIN-MEDIATED DRUG-HYPACONITINE INTERACTIONS BY CACO-2 HUMAN INTESTINAL CELL MONOLAYERS

Aconitum species is an essential but toxic herb in Traditional Chinese Medicine (TCM). As a main and characteristic constituent in Aconitum species, hypaconitine (HA) was selected to illustrate the P-glycoprotein-mediated drug-HA interactions using Caco-2 cell monolayers. The HA absorption results revealed that HA was not only an inhibiter but also a substance of P-glycoprotein (P-gp). In TCM theory, it is acknowledged that Aconitum species are incompatible with Bulbus Fritillariae for the increase of the poisoning risk. Peimine and peiminine, which are the active components of Bulbus Fritillariae, increased the bioavailability of HA suggested this effect should be one of the reason to increase the poisoning risk when Bulbus Fritillariae combined with Aconitum species. However, no significant difference in absorption of peimine (PE) or peiminine (PEN) was observed when they combined with HA, respectively. Glycyrrhiza is a famous detoxifying herb in TCM, and the combined use of Glycyrrhiza and Aconitum species is a safe pair in TCM formulations. Liquiritin (LQ) and isoliquiritin (ILQ), which are the active components of Glycyrrhiza, were combined with HA, respectively. The detoxified mechanism of Glycyrrhiza combined with Aconitum species seems less related to the interaction of the absorption.

Two new pyrazines from the parent roots of Aconitum carmichaelii

Two new pyrazines, aconicarpyrazine A (1) and aconicarpyrazine B (2), together with five known heterocyclic compounds: adenosine (3), uridine (4), hypoxanthine (5), nicotinamide (6), and uracil (7), were isolated from the parent roots of Aconitum carmichaelii. The structures of these alkaloids were elucidated by spectroscopic analysis, including 2D NMR techniques. This is the first report of pyrazines in a species of Aconitum.

Aconitum rotundocassideum , a New Species of Ranunculaceae from Shaanxi

A new species of the genus Aconitum( Ranunculaceae), Aconitum rotundocas-sideum,is described here from Shaanxi Province. Having similar habit and galeiform upper sepals,this species is closely related to A. crassicaule W. T. Wang endemic to Gongshan County,Yunnan Province,though the latter differs by its flower arrangement in a panicle with three small 3-flowered racemes, usually arcuate pedicels, smaller roundedgaleiform or subglobose,conspicuously nerved,not beaked upper sepal and smaller lateral sepals,and 2-denticulate stamen filaments.

Specific detection of Japanese aconite using multiplex PCR

Aconite is one of the most toxic plants, and is known world-wide. To confirm cases of poisoning by aconite, it would be helpful to be able to identify aconite DNA from samples such as stomach contents. In this study, we designed a tetraplex PCR targeted to the ITS region of the aconite nuclear DNA and three loci (matK, trnL-F and psbA-trnH) in chloroplast DNA. Four amplification products were specifically detected from all Japanese aconite samples we tested, substances with suspected aconite contamination, and a medicine, as well as in highly degraded materials. Furthermore, the sequencing analysis of psbA-trnH revealed that seven types of aconite were observed in Japanese samples. This method is a simple, sensitive and reliable method that can be used to identify Aconitum species.

UPLC–Q‐TOF–HDMS Analysis of Constituents in the Root of Two Kinds of Aconitum Using a Metabolomics Approach

Metabolomics is an 'omics' approach that aims to comprehensively analyse all metabolites in a biological sample, and has great potential for directly elucidating plant metabolic processes. Increasing evidence supports the view that plants produce a broad range of low-molecular-weight secondary metabolites responsible for variation from species to species, thus enabling the use of secondary metabolite profiling in the chemotaxonomy. To gain deeper insights into the metabolites to increasing plant diversity, we performed systematic untargeted metabolite profiling to exploit the different parts and species of Aconitum as a case study. Application of ultraperformance liquid chromatography-quadrupole time-of-flight-high-definition mass spectrometry (UPLC-QTOF-HDMS) equipped with electrospray ionisation and coupled with pattern recognition analyses to study constituents in the root of two kinds of Aconitum species. Twenty-two metabolites between the mother root of Aconitum carmichaelii Debx (CHW) and lateral root of Aconitum carmichaelii Debx (SFZ) and 13 metabolites between the CHW and root of Aconitum kusnezoffii Reichb (CW) have been identified. Of note, songorine, carmichaeline and isotalatizidine did not exist in CW, whereas they are present in the SFZ and CHW. Metabolomics based UPLC-QTOF-HDMS with multivariate statistical models was effective for analysis of constituents in the root of two kinds of Aconitum species.

The exposure of highly toxic aconitine does not significantly impact the activity and expression of cytochrome P450 3A in rats determined by a novel ultra performance liquid chromatography–tandem mass spectrometric method of a specific probe buspirone

Aconitum species are widely used to treat rheumatism, cardiovascular diseases, and tumors in China and other Asian countries. The herbs are always used with drugs such as paclitaxel. Aconitine (AC) is one of the main bioactive/high-toxic alkaloids of Aconitum roots. AC is metabolized by cytochrome P450 (CYP) 3A. However, whether AC inhibits/induces CYP3A, which causes drug–drug interaction (DDI) is unclear. Our study aims to explore the potent effects of AC, as a marker component of Aconitum, on CYP3A using the probe buspirone in rats. The effects of oral AC on pharmacokinetics of buspirone were evaluated. CYP3A activity and protein levels in rat liver microsomes pretreated with oral AC were also measured using in vitro buspirone metabolism and Western blot. Buspirone and its major metabolites 1-(2-pyrimidinyl)piperazine and 6′-hydroxybuspirone were determined using a newly validated UPLC–MS/MS method. Single dose and 7-day AC administration at 0.125mg/kg had no effect on CYP3A activity since no change in the formation of 1-(2-pyrimidinyl)piperazine and 6′-hydroxybuspirone. CYP3A activity and protein levels in liver microsomes were also not affected by 7-day AC pretreatment at 0.125mg/kg. Therefore, AC neither inhibits nor induces CYP3A in rats, indicating AC does not cause CYP3A-related DDI in the liver.

Genetic diversity assessment of Aconitum coreanum (H. Lév.) Rapaics (Ranunculaceae), an endangered plant species in Korea, using microsatellite markers

To assess the genetic diversity of Aconitum coreanum (Ranunculaceae) populations in Korea, we have amplified and sequenced eight organellar marker regions, and developed and analyzed microsatellite markers. No sequence variation was detected from the eight organellar markers. Ten microsatellites were developed using Next Generation Sequencing and two microsatellite markers, AK_CA03 and AK_CT07, were identified polymorphic and applied for 143 individuals of twelve A. coreanum populations. Four and five alleles were detected for the two microsatellite loci, respectively, and number of migrants (<TEX>$N_m$</TEX>) was estimated as 1.12586. Two microsatellite marker loci showed <TEX>$F_{ST}$</TEX> of 0.205 and 0.275, respectively. The heterozygosity deficit, low level of among-population differentiation, small size of gene flow, and lack of sequence variation of the organellar markers suggest that A. coreanum is reproductively isolated from other Aconitum species and there has been continuous gene flow among the populations of A. coreanum or it has dispersed relatively recently after speciation. Though population pairwise <TEX>$F_{ST}$</TEX>'s presented significant geographic structure, further sampling and study will be necessary to confirm this.

Phytochemicals, Traditional Uses and Processing of Aconitum Species in Nepal

The tuberous roots of genus Aconitum are commonly applied for various diseases, such as rheumatic fever, painful joints and some endocrinal disorders. It stimulates the tip of sensory nerve fibers. These tubers of Aconitum are used in the herbal medicines only after processing. At the mean time, there remain high toxicological risks of the improper usages of Aconitum. The cardio- and neurotoxicity of this herb are potentially lethal. Some of the species like A. ferox and A. spicatum are deadly poisonous while others like A. oreochryseum and A. bisma are used as the antidote for Aconitum poisoning. It is therefore, a valuable drug as well as an unpredictable toxic material. In the current review, assessments of Aconitum species are carried out, to increase knowledge for the safety uses, in context of Nepal. The traditional uses, phytochemical studies, its processing techniques and toxicological principles are reviewed.DOI: http://dx.doi.org/10.3126/njst.v12i0.6496 Nepal Journal of Science and Technology 12 (2011) 171-178

Aconitum balfourii Stapf: A rare medicinal herb from Himalayan Alpine

Aconitum is a genus of flowering plant belonging to buttercup family (Ranunculaceae). Globally, there are over 300 species of Aconitum. In India, the genus is represented by about 24 species mainly distributed in subalpine and alpine zones of Himalayas. Out of these species, Aconitum balfourii Stapf, known as Mitha and Vatsnabh, is a significant species of this genus. It is widespread in Kumaon and Garhwal Himalayas on shady slopes from 3000 to 4200 m altitudes. Tuberous roots of A. balfourii Stapf are rich sources of pseudoaconitine (0.4 to 0.5%) and aconite alkaloids. The value of aconite as a medicine has been fully recognized in modern times, and it now ranks as one of the most useful drugs, particularly in Homeopathy, Ayurveda, and Unani systems of medicine. Due to overexploitation, A. balfourii Stapf is facing severe threat, and the plant has been listed among 37 Himalayan medicinal herbs under priority for in situ and ex situ conservation. This review mainly discussed several aspects, such as: distribution, cultivation, morphological characteristics, basis of origin, and conservation of this important species of the genus Aconitum. Key words: Aconitum, conservation, evolution, propagation, pseudoaconitine.

Bombus gerstaeckeri Morawitz, 1881 (Hymenoptera, Apidae) : observations sur la biologie d’un bourdon localisé et oligolectique

Bombus gerstaeckeri Morawitz, 1881 (Hymenoptera, Apidae): observations on the biology of a localized and oligolectic bumblebee. - Bombus gerstaeckeri Morawitz 1881 is a particular species for its scarcity and oligolecty on species of the genus Aconitum (Ranunculaceae). We performed a sampling of B. gerstaeckeri across its entire distribution range during the summers of 2010 and 2011, in the framework of a study on the evolution of floral specialisation with a phylogeographic approach in the subgenus Megabombus. Here we present recent observations during our field surveys.

Assessment of the toxicity of two Aconitum herbal medicines by microcalorimetry and chemometrics

In this study, a non-invasive and non-destructive microcalorimetric method was applied to assess the toxic effects of two Aconitum herbal medicines, Radix Aconitum Kusnezoffii and Radix A. carmichaeli, on the multiplying and non-multiplying metabolism of Shigella dysenteriae. Some crucial information, such as the thermogenic curves and thermo-kinetic parameters including growth rate constant k, inhibitory ratio I and half-inhibitory concentration IC50, of S. dysenteriae growth affected by the two Aconitum species was obtained. Coupled with chemometric analysis on this information, it could be found that Radix A. Kusnezoffii and Radix A. carmichaeli could be differentiated according to their toxic effects; Radix A. Kusnezoffii with IC50 of 35.2mg/mL had stronger toxic effect than Radix A. carmichaeli with IC50 of 49.6mg/mL. This present study shows the effectiveness of combining microcalorimetry and chemometrics for assessing the toxic effects of herbal medicines and other food matrices.

Consolida and Aconitella are an annual clade of Delphinium (Ranunculaceae) that diversified in the Mediterranean basin and the Irano‐Turanian region

AbstractThe Mediterranean and Irano‐Turanian biogeographic regions are biodiversity hotspots for mesophytic and xerophytic species, including many Delphinieae. This phylogenetically poorly understood tribe of Ranunculaceae consists of Consolida and Aconitella, with together ca. 52 species, and Delphinium and Aconitum, each with ca. 300 species. To infer the phylogeny of Consolida and Aconitella, we analyzed nuclear and chloroplast DNA sequences from 39 of their species and subspecies (44 taxa) plus a set of 30 exemplar species of Delphinium and Aconitum. We used a Bayesian relaxed clock model to estimate divergence times and a maximum likelihood approach to reconstruct ancestral areas. Aconitella forms a clade embedded in Consolida, and the latter is embedded in Delphinium. Consolida s.l. (including Aconitella) comprises two clades in the Irano‐ Turanian region and three in the Mediterranean basin. The latter clades’ inferred crown ages of 5.1, 4.4, and 2.8 Ma suggests that the repeated drying‐up of the Mediterranean, concomitant with and following the Messinian salinity crisis (5.96–5.33 Ma), may have facilitated their westward expansion. While there is clear geographic structure towards the tips of the Consolida s.l. tree, a likely ancestral area could not be inferred. However, the initial diversification of Consolida s.l., which occurred ca. 17 Ma ago, falls in a period when the climate in the Anatolian region became more arid, which may have favoured the annual life cycle that characterizes all species in this clade. To achieve a classification of mutually monophyletic genera in Delphinieae may require transferring the species of Aconitella and Consolida into Delphinium.

Antioxidant and immunological activities of water-soluble polysaccharides from Aconitum kusnezoffii Reichb.

Aconitum kusnezoffii Reichb., one of the earliest recorded toxic species of genus Aconitum, has been used as traditional Chinese medicine and medicinal diet over the last 2500 years to treat heart failure congestion, neuralgia, rheumatism and gout, etc. In the present paper, four water-soluble polysaccharide fractions isolated from the tubers of A. kusnezoffii Reichb. were studied the antioxidant and immunological activities for the first time. In vitro antioxidant assays indicated that fraction WKCP-A had noticeable scavenging activities on DPPH radical, hydroxyl radical, superoxide anion, H2O2 and self-oxidation of 1,2,3-phentriol, ferrous ion-chelating ability and reducing power. Moreover, the in vivo immunological assay exhibited that fractions WKCP-A and WKHP could more significantly enhance splenic lymphocyte proliferation and macrophage phagocytosis than other fractions. Therefore, the water-soluble polysaccharides from A. kusnezoffii Reichb., especially WKCP-A, have the potential to be explored as novel natural antioxidants and immunostimulating agents for using in functional foods or medicine.

Seven cases of fatal aconite poisoning: Forensic experience in China

This paper presents seven fatal cases of aconite poisoning encountered in the Tongji Center for Medicolegal Expertise in Hubei (TCMEH), China, from 1999 to 2008 retrospectively. In six of the cases, deaths occurred after drinking homemade medicated liquor containing aconite, and in one case death was due to ingestion of traditional Chinese medication containing aconite. Forensic autopsy and pathological examinations ruled out the presence of physical trauma or life-threatening diseases. Diagnosis of aconite poisoning was made after postmortem toxicological analysis. Animal experiment was performed in one case demonstrating that the medicated liquor could cause death rapidly. We present the autopsy and histopathological findings, toxicological analysis, and results of animal experiment done on samples from those seven cases. As an important herbal Chinese medicine, Aconitum species deserve special attention, especially because it contains poisonous alkaloids.

Microsomal cytochrome P450-mediated metabolism of hypaconitine, an active and highly toxic constituent derived from Aconitum species

Hypaconitine (HA), an active and highly toxic constituent derived from Aconitum species, is widely used to treat rheumatism. Little is known about the hepatic cytochrome P450-catalyzed metabolism of HA. The present study investigated the metabolism of HA in vitro using male human liver microsomes (MHLMS). Chemical inhibitors of specific CYP enzymes, CYP-specific inhibitory monoclonal antibodies (mAbs), and cDNA-expressed CYP enzymes were used to confirm the enzyme subtypes involved in the metabolism. Liquid chromatography–high resolution mass spectrometry (LC–MS) was used to detect and identify metabolites. A total of 11 metabolites were identified in MHLMS incubations. The major metabolic pathways included demethylation (M1–M3), demethylation–dehydrogenation (M4–M6), hydroxylation (M7, M8), and didemethylation (M9–M11). M8 was identified as mesaconitine (MA), another active and highly toxic constituent of Aconitum. The results of chemical inhibition, monoclonal antibody inhibition, and cDNA-expressed CYP enzyme studies showed that the primary contributors toward HA metabolism were CYP3A4 and 3A5, with secondary contributions by CYP2C19, 2D6, and CYP2E1. CYP1A2 and 2C8 provided minor contributions.

Alkaloids from two species of the genus Aconitum

We investigated the aerial part of Aconitum leucostomum Worosch collected in the Altai Mountains of Xinjiang– Uyghur Autonomous Region of the PRC. Extraction by CHCl3 of the aerial part of the plant (2.9 kg) afforded total bases (13.26 g, 0.45%). Part of the total alkaloids (6.7 g, pH 6) was chromatographed over a column of Al2O3 with elution by hexane:acetone (5:1, 1:1), CHCl3, and CHCl3:MeOH (5:1, 1:1). The first hexane:acetone effluents afforded lappaconitine (2.09 g) and N-deacetyllappaconitine (0.13 g). Part of the total bases (6.36 g, pH 10) was chromatographed over a column of Al2O3 with elution by hexane:acetone (5:1, 1:1), acetone, and acetone:MeOH (10:1, 5:1, 1:1). The hexane:acetone effluents afforded lappaconitine (1.16 g) and ranaconitine (0.06 g). Base 1 (0.04 g), mp 234°C, was isolated from the acetone fractions. In continuation of the investigation of the alkaloid composition of A. septentrionale (Ranunculaceae) roots collected in the Russian Federation in the Republic of Bashkortostan, we chromatographed the mother liquor of lappaconitine (64 g) over a column of silica gel (290 g) [1]. The column was eluted with CHCl3 and CHCl3:MeOH (100:1, 50:1, 25:1) A total of 460 100-mL fractions was collected. Work up of the initial CHCl3 effluents (fractions 1–5) with acetone separated lappaconitine (2.66 g), mp 216–218°C; of subsequent fractions (6–35), N-deacetyllappaconitine (0.18 g), mp 208–211°C. The CHCl3:MeOH effluents (100:1) (fractions 131–151) were worked up with acetone:EtOH to afford sepaconitine (0.06 g), mp 253–255°C. The CHCl3:MeOH effluents (50:1) (fractions 190–206) were rechromatographed over a column of silica gel. Elution by CHCl3 and CHCl3:MeOH (50:1) isolated songorine (0.02 g), mp 201–203°C. Then, CHCl3:MeOH (25:1) gave fractions 385–425 that were rechromatographed over a column of silica gel with elution by CHCl3:MeOH (50:1 and 25:1). Fractions of 10 mL were collected. Fractions that were identical with respect to Rf were combined and worked up with EtOH to isolate 1, C22H33NO2, mp 236°C (EtOH).

Рід Aconitum L. в Українських Карпатах

The key for identification, the maps of distribution, sozological evaluation, critical taxonomical notes and morphological descriptions of monkshood species from the Ukrainian Carpathians are presented. It is ascertained that in this territory there are 22 taxa (12 species) of genus Aconitum L. which belong to 5 sections and 3 subgenera – section Lycoctonum DC. of the subgenus Lycoctonum (DC.) Peterm., section Anthora DC. of the subgenus Anthora (DC.) Peterm., and sections Aconitum , Cammarum DC., Acomarum Starmühl. of the subgenus A conitum . Keywords: Aconitum, morphology, taxonomical key, chorology.

Toxicological Risks of Chinese Herbs

As traditional Chinese medicine (TCM) has become more popular there have been increasing concerns about safety and potential toxicity of the Chinese materia medica (CMM) comprising plants, animal parts and minerals. The potential toxicity of many CMM is well recognised in TCM and to reduce risks use of some herbs is restricted whilst specific processing methods have been developed to modify the activities/toxicity of others. However adverse reactions have been reported, many of these are due misuse or abuse of Chinese medicine. The main problem remains products adulterated with pharmaceuticals for weight loss or erectile dysfunction. But some herbs have narrow therapeutic ranges (e.g., Aconitum species) so toxic effects are frequently reported. Toxic effects from chronic or cumulative dosing are difficult to detect in the traditional setting and recent reports have demonstrated the health problems from Aristolochia species. Despite safety concerns, Chinese medicine appears to be relatively safe with comparatively few reports of adverse reactions compared with overall drug reports. The wealth of information in the Chinese literature needs to be more widely available. As TCM is widely used by patients, improved pharmacovigilance and pharmacoepidemiology can contribute valuable safety information, relevant to clinical use.

Identification of the Medicinal Plants inAconitumL. by DNA Barcoding Technique

Plants of the genus Aconitum L. are commonly used in Asia for medicinal purposes. Although they are widely cultivated and marketed, there has been uncertainty about the efficacy of different species, and therefore accurate identification is crucial. To determine the genetic variation among these medicinal plants, the proposed DNA barcode PSBA- TRNH intergenic spacer of 134 individuals from 19 taxa of ACONITUM were sequenced. Among the two most commonly used medicinal ACONITUM species, A. carmichaeli and A. kusnezoffii, sequence inversions were observed. The studied samples were clustered into ten groups according to the sequence alignment and most of the tested Aconitum species could be differentiated by the PSBA -TRNH intergenic spacer.