Molecular identification of Cynanchum auriculatum and its closely related species based on ITS2 sequence / 中草药

Objective To identify Cynanchum auriculatum and its closely related species using the ITS2 barcode. Methods A total of 54 samples of C. auriculatum and its related species were collected. Genomic DNAs were extracted from these samples. The ITS2 sequences of these samples were amplified and bidirectional sequenced by PCR. The obtained sequences were submitted to the Gen Bank and the ITS2 sequences of 47 samples belonging to 15 species were downloaded from the GenBank, and ITS2 sequences were annotated by ITS2 database. A total of 101 ITS2 sequences were aligned and the intraspecific and interspecific distances were calculated using the MEGA 5.0. Identification analyses were performed using the similarity search method and nearest distance method, and were presented intuitively by constructing neighbor-joining (NJ) tree. Results The length of all ITS2 sequences of C. auriculatum was 249 bp, which was a haplotype and was close to Cynanchum. There was a significant difference between the interspecific and intraspecific genetic distances of the ITS2 sequences. The NJ tree showed that C. auriculatum obviously differed from its closely related species, which showed high monophyly. According to the secondary structure of ITS2, it was also possible to distinguish between C. auriculatumi and Asclepiadaceae related species. Conclusion As a DNA barcode, ITS2 sequences can stably and accurately distinguish C. auriculatum from its closely related species and also provide a new technique to ensure the clinical safety in utilization of Chinese materia medica.

Paniculatumoside G, a new C21 steroidal glycoside from Cynanchum paniculatum

ABSTRACT A new C21 steroidal glycoside, paniculatumoside G, together with neocynapanogenin C isolated for the first time from the natural source and two known compounds were isolated and characterized from the roots and rhizomes of Cynanchum paniculatum (Bunge) Kitag. ex H.Hara, Apocynaceae, a commonly used Traditional Chinese Medicine. On the basis of spectroscopic analysis, including HR-ESI-MS, 1D and 2D NMR spectral data, the structure of the new C21 steroidal glycoside was elucidated as neocynapanogenin H 3-O-β-D-oleandropyranoside.

Recent advances in phytochemistry and pharmacology of C21 steroid constituents from Cynanchum plants / 中国天然药物

Cynanchum is one of the most important genera in Asclepiadaceae family, which has long been known for its therapeutic effects. In this genus, 16 species are of high medicinal value. The extracts of the root and/or rhizome parts have been applied in traditional Chinese medicines (TCM) for the prevention and treatment of various illnesses for centuries. C21 steroids, as the typical constituents of Cynanchum species, possess a variety of structures and pharmacological activities. This review summarizes the comprehensive information on phytochemistry and pharmacology of C21 steroid constituents from Cynanchum plants, based on reports published between 2007 and 2015. Our aim is to provide a rationale for their therapeutic application, and to discuss the future trends in research and development of these compounds. A total of 172 newly identified compounds are reviewed according to their structural classifications. Their in vitro and in vivo pharmacological studies are also reviewed and discussed, focusing on antitumor, antidepressant, antifungal, antitaging, Na(+)/K(+)-ATPase inhibitory, appetite suppressing and antiviral activities. Future research efforts should concentrate on in vitro and in vivo biological studies and structure activity relationship of various C21 steroid constituents.

Three New C21 Steroidal Glycosides from Roots of Cynanchum otophyllum / 中草药·英文版

Objective: To investigate the chemical structures of glycosides in the roots of Cynanchum otophyllum (Asclepiadaceae) and to find new glycosides. Methods: The total glycosides in the roots of C. olophyllum were separated by silica gel column chromatography. The structures of the resulting compounds were determined by NMR and FAB-MS spectra. Results: Three C21 steroidal glycosides were separated. Their structures were determined as caudatin 3-O-(4-O-methyl-β-D-cymaropyranosyl)-(1→4)-α-D-oleandropyranosyl-(1→4)-β-D-glucopyranosyl-(1→4)-α-L-rhamnopyranoside (1), caudatin 3-O-β-D-digitoxopyranosyl-(1→4)-α-D-oleandropyranosyl-(1→4)-β-Ddiginopyranosyl-(1→4)-β-D-glucopyranoside (2), and caudatin 3-O-β-D-diginopyranosyl-(1→4)-α-D-oleandropyranosyl-(1→4)-α-D-oleandropyranosyl-(1→4)-β-D-glucopyranoside (3), respectively. Conclusion: Glycosides 1-3 are new compounds.

Chemical constituents of Calotropis gigantea L. and their anticancer activity / 中国药学杂志

OBJECTIVE: To study the anti-cancer components of Calotropis gigantean L. METHODS: The powdered whole plants of C. gigantea were extracted with 95% alcohol. After removal of the solvent, the residue was extracted with petroleum ether and chloroform, and the compounds in the chloroform extract were isolated and purified by different column chromatograghies carried out on silica gel, RP-18, MCI, and Sephadex LH - 20 and their structures were elucidated by spectral data. RESULTS: Thirteen compounds were isolated and their structures were characterized as gofruside(1), uzarigenin(2), arjunolic acid(3), 3ξ-(1ξ-hydroxyethyl) -7-hydroxy-1-isobenzofuranone(4), daucosterol(5), syringaresinol(6), 12-O-benzoyl-deacylmetaplexigenin(7), 3-hydroxy-4-methoxybenzoic acid(8), oleanolic acid(9), β-sitosterol(10), methyl 1-naphthaleneacetate(11), butylparaben(12), α-D-oleandropyranoside(13), and compounds 2 - 4, 6- 9, and 11 - 13 were isolated from this plant for the first time. Compounds 1 and 2 showed cytotoxicity against HLE, K562, RPMI8226, MCF7, MDA, and WM9 cell lines, with K562 and RPMI8226 being the most sensitive cells. CONCLUSION: Compounds 1 and 2 are premilinarily judged as the anti-cancer components in C. gigantean.

C21 Steroidal Saponins from Cynanchum otophyllum / 中草药·英文版

Objective: To study the chemical structures of saponins in the rhizome of Cynanchum otophyllum (Asclepiadaceae), and to find new compounds. Methods: The total saponins of the rhizome were separated by silica gel column chromatography. The chemical structures of obtained compounds were determined by NMR and FAB-MS spectra. Results: Three C21 steroidal saponins were separated. Their structures were determined as caudatin 3-O-β-D-glucopyranosyl-(1→4)-β-D-digitoxopyranosyl-(1→40)-β-D-digino-pyranosyl-(1→4)-α-D-oleandropyranoside (1), caudatin 3-O-β-D-oleandropyranosyl-(1→4)-α-D-oleandropyranosyl-(1→4)-α-D-oleandropyranoside (2), and caudatin 3-O-β-D-glucopyranosyl-(1→4)-α-D-oleandropyranosyl-(1→4)-β-D-diginopyranosyl-(1→4)-α-D-oleandropyranoside (3), respectively. Conclusion: Saponins 1-3 are new compounds.

Development of Simple, Cost Effective Protocol for Micropropagation of Tylophora indica (Burm f.) Merill., an Important Medicinal Plant.

Aims: The present studies were initiated to develop a cost effective protocol for micropropagation, as a mean for conservation of medicinal plant- Tylophora indica (Burm f.) Merill. The plant is threatened and needs immediate conservation, therefore, the study was undertaken with following objective:  In vitro multiplication of Tylophora indica using nodal axillary bud proliferation and through organogenesis of callus. Study Design: For all experiments ten replicates were used per treatment and all the experiments were repeated three times. Data have been presented as Mean ± Standard deviation. Place and Duration of Study: Department of Plant Cell and Molecular Biology, Indian Institute of Advanced Research (IIAR), Gandhinagar, Gujarat, India. Methodology: For in vitro plant regeneration, micropropagation and organogenesis techniques were used. For micropropagation, surface sterilized nodal explants were inoculated on different shoot inducing media and further multiplication was obtained. Root containing shoots were transferred to pot containing pre autoclaved mixture of soil and soilrite. For organogenesis, surface sterilized leaf explants were inoculated on different types of callus inducing media. Results: All the nodal explants were sprouted at a very high frequency, i.e. 98% and sprouted buds elongated up to 8cm on three different media. Dissected explants grew further and average height of shoots reached 9.5cm±0.80cm within 30 days. Interestingly, root formation was observed on the same media; so that the best media was 0.4mg L-1 BA and 0.1mg L-1 Kn for both initiation as well as for multiplication. For organogenesis, the fragile callus was observed on media containing 2mg L-1 2,4-D and 0.1mg L-1 Kn. Green pigmented calli were transferred to MS media, where it regenerated in to shoots and roots, simultaneously Conclusion: The protocol of micropropagation through axillary bud proliferation described here is very simple, repetitive and cost effective, which can be easily utilized for commercial cultivation. On shoot multiplication media, root formation observed, thereby making the process is one step; which is very easy to follow.

Phytochemistry, pharmacology and traditional uses of Leptadenia pyrotechnica- An important medicinal plant.

Traditional Systems of medicines always played important role in the global health. In the traditional health medicinal plants providing a new areas of drug research. The demand for plant based medicines, food supplement, health products, pharmaceuticals and cosmetics are increasing in both developing and developed countries due to the growing recognition that the natural products are non toxic, have less side effects and easily available. Leptadenia pyrotechnica (Forsk.) commonly known as Kheep belonging to family Asclepiadaceae. Leptadenia pyrotechnica leafless much branched shrub. All parts of Leptadenia pyrotechnica are used in traditional medicines. The present article gives an account of updated information on its phytochemistry pharmacological properties. Ethnomedical uses say to possess significant antioxidant, anti-inflammatory, antibacterial, anthelmentic, antilipoxygenase, cytotoxic, antitumour, hypolipidemic and anti atherosclerotic activity. The present review contains wide number of isolated chemical constituents and various ethnomedical and traditional uses of Leptadenia pyrotechnica. It include information about historical background, conceptual basis, different disciplines studied in the systems, research and development aspects, drug manufacturing aspects and impact of globalization.

C21 Steroidal Glycosides from Acidic Hydrolysate of Cynanchum otophyllum / 中草药·英文版

Objective: To investigate the structures of compounds in the rhizome of Cynanchum otophyllum (Asclepiadaceae), and to find new C21 steroidal glycosides. Methods: The ethyl acetate extract from the rhizome was subjected to acidic hydrolysis and isolated by column chromatography; The structures of the purified compounds were determined by spectral methods. Literature search confirmed whether those compounds were of new structures. Results: Three compounds were isolated and their structures were deacetylmetaplexigenin 3-. O-β-. D-oleandropyranosyl-(1→4)-α-. D-oleandropyranosyl-(1→4)-α-. D-oleandropyranoside (1), deacetylmetaplexigenin 3-. O-α-. D-oleandropyranosyl-(1→4)-β-. D-thevetopyranosyl-(1→4)-α-. D-oleandropyranoside (2), and deacetylmetaplexigenin 3-. O-β-. D-cymaropyranosyl-(1→4)-α-. D-oleandropyranoside (3), respectively. Conclusion: Compounds 1-3 are new compounds.

Saponins-rich fraction of Calotropis procera leaves elicit no antitrypanosomal activity in a rat model

Objective:To examine the in vitro and in vivo anti-Trypanosoma evansi (T. evansi ) activity of saponins-rich fraction of Calotropis procera (cpsf) leaves as well as the effect of the fraction on the parasite-induced anemia. Methods:A 60-minutes time course experiment was conducted with various concentrations of the fraction using a 96-well microtiter plate technique, and subsequently used to treat experimentally T. evansi infected rats at 100 and 200 mg/kg body weight. Index of anemia was analyzed in all animals during the experiment. Results:The cpsf did not demonstrate an in vitro antitrypanosomal activity. Further, the cpsf treatments did not significantly (P>0.05) keep the parasites lower than the infected untreated groups. At the end of the experiment, all T. evansi infected rats developed anemia whose severity was not significantly (P>0.05) ameliorated by the cpsf treatment. Conclusions:It was concluded that saponins derived from Calotropis procera leaves could not elicit in vitro and in vivo activities against T. evansi.

Gastric antiulcer and antiinflammatory activities of Calotropis procera stem bark

In recent years, a widespread search has been launched to identify new antiinflammatory and antiulcer-drugs from natural sources. The study was aimed at evaluating the antiinflammatory and antiulcer activity of chloroform extract (CH) and hydroalcoholic extract (HE) of the stem bark of Calotropis procera (Aiton) W.T. Aiton, Apocynaceae, obtained successively by cold maceration. The antiinflammatory effect of the CH and HE extracts of the stem bark of the C. procera against carrageenan-induced paw oedema and also its antiulcer activity by using two acute models: Aspirin (100 mg/kg, p.o.) and ethanol (96 percent, 1 mL/200 g) in albino rats have been studied and found to be significant at 200 and 400 mg/kg when compared to the standard drugs. As a part of investigations to obtain compounds with antiinflammatory and antiulcer activity in this work, a bioassay was carried out with fractions obtained from chloroform extract with n-hexane (NF1), 1-butanol (BF1), ethyl acetate (EF1) and chloroform (CF1). The hydroalcoholic extract (HE) of the stem bark was fractionated with n-hexane (NF2), 1-butanol (BF2), ethyl acetate (EF2), chloroform (CF2) and water (WF2). The fractions were freeze-dried and evaluated for its antiinflammatory and antiulcer activity. Fractions NF1, CF1, BF2 and EF2 (20 mg/kg) showed significant antiinflammatory and antiulcer activity. The results obtained for antiulcer activity were also supported well by the histopathological examination of the open excised rat stomach. Further experiments are underway to determine which phytoconstituents are involved in antiinflammatory and antiulcer activities as well as mechanisms involved in gastroprotection.

Diagnose of the ingestion of Asclepias mellodora St. Hil. by sheep through microhistological analysis of their digestive contents / Diagnose da ingestão de Asclepias mellodora St. Hil. por ovinos através da análise micro-histológica de seu conteúdo digestivo

Asclepias mellodora St. Hil. is a native acute toxic species frequent in the grasslands of the Buenos Aires province, Argentina, whose toxicity had not been assessed until now. This study evaluates the minimal lethal dose of this species for sheep, and the possibility of microscopically recognizing its fragments in gastrointestinal contents as a complementary diagnostic tool in necropsies. Three Frisona sheep (average LW=55±4.5 kg) were dosed via an esophageal tube with each one of the following doses of asclepias: 8.0, 5.0, 2.0 and 0.8 g DM.kg LW-1. Sheep poisoned with the three higher doses died between 10 and 85 h after intoxication, but those receiving the lower dose did not. During necropsies we: 1) determined the dry weight of the contents of rumen+reticulum, omasum+abomasum, and large intestine, 2) estimated the percentages of asclepias fragments by microanalysis correcting for digestion effects on fragment recognition, and 3) calculated the total mass of asclepias in the digestive tract of each animal. For the three higher doses, the mass of asclepias identified in the total ingesta was 12.3±3.4 percent of the amount supplied, possibly because of the strong diarrhea its ingestion produced. The percentages of asclepias in rumen+reticulum did not differ from the average quantified for the entire tract. The results of this study indicate that the minimal lethal doses of asclepias for sheep is between 2.0 and 0.8g DMÀkg LW-1, and that the microhistological analysis of the rumen+reticulum, the easiest region to sample, can be used to confirm the ingestion of this toxic species, although the estimated percentage will be not a good estimator of the ingested percentage.

Asclepias mellodora St. Hil. é uma espécie nativa de aguda toxicidade, frequente nos campos da província de Buenos Aires, Argentina. A sua toxicidade não foi avaliada até agora. Este estudo avalia a dose mínima letal desta espécie, para os ovinos, bem como a possibilidade de reconhecer microscopicamente seus fragmentos no conteúdo gastrointestinal como uma ferramenta complementar de diagnóstico em necropsias. Três ovinos Frisona (PV média = 55±4,5 kg) foram dosados através de uma sonda esofágica em cada uma das seguintes doses de Asclepias: 8,0, 5,0, 2,0 e 0,8 g DM.kg PV-1. Ovinos intoxicados com as três maiores doses morreram entre 10-85 h após a intoxicação, mas não aqueles que receberam a dose menor. Durante as necropsias se: 1) determinou o peso seco do conteúdo do rúmen + retículo, omaso + abomaso e intestino grosso, 2) estimou as porcentagens de fragmentos de Asclepias por microanálise, fazendo a correção para efeitos de digestão no reconhecimento dos fragmentos, e 3) calculou a massa total de Asclepias no trato digestivo de cada animal. Para as três doses maiores, a massa de Asclepias identificada na ingesta total foi de 12,3±3,4 por cento da quantidade fornecida, possivelmente por causa da forte diarréia produzida pela sua ingestão. As porcentagens de Asclepias no rúmen + retículo não diferiram da média quantificada para o trato completo. Os resultados deste estudo indicam que a dose letal mínima de Asclepias em ovinos é de entre 2,0 e 0,8 g kg PV ò DM-1, e que a análise micro-histológica do rúmen + retículo, a região mais fácil de amostrar, pode ser usada para confirmar a ingestão desta espécie tóxica, embora a percentagem estimada não será um bom estimador da porcentagem ingerida.

Asclepiadaceae endémicas del Perú

La familia Asclepiadaceae es reconocida en el Perú por presentar 27 géneros y 107 especies (Brako & Zarucchi, 1993), mayormente bejucos y hierbas. Esta familia se halla bajo revisión sistemática. Para la flora peruana, estos estudios en desarrollo probablemente concluyan con la necesidad de un reordenamiento de los taxones a nivel de los géneros (S. Liede, com. pers.). Dada la escasez de colecciones herborizadas y la falta de consenso sobre los taxones, aquí se reconocen provisionalmente 43 endemismos. En general, los endemismos en esta familia ocupan las regiones Mesoandina y Bosques Muy Húmedos Montanos, entre los 900 y 2600 m de altitud. Se aplicaron las categorías y criterios de la UICN a cinco especies. Dos endémicas se encuentran registradas en el Sistema Nacional de Áreas Naturales Protegidas por el Estado.

The Asclepiadaceae are represented in Peru by 27 genera and 107species (Brako & Zarucchi, 1993), mostly vines and lianas. The family is under systematic revision, and these ongoing studies are likely to result in genus-level changes for the Peruvian Asclepiadaceae flora (S. Liede, pers. comm.). Given the scarcity of collections and the lack of consensus on the taxonomic status of several taxa, we provisionally recognize 43 endemic species. The endemic taxa are found in the Mesoandean and Very Humid Montane Forest regions, between 900 and 2600 m elevation. We applied IUCN categories and criteria to five species. Two endemic species have been found in protected areas.

Chemical Compositions and Pharmacological Activities of Asclepiadaceae Plants / 浙江中医药大学学报

The present paper summarizes recent studies of chemical compositions and pharmacological activities of Asclepiadaceae plants.

Aprovechamiento de Morrenia odorata (Hook & Arn.) Lindl. (Asclepiadeceae) «TASI¼ en medicina tradicional como una alternativa en la alimentación humana / Use of Morrenia odorata (Hook & Arn.) Lindl. (Asclepiadeceae) «TASI¼ in traditional medicine as an alternative in human nutrition

Es importante la revalorización de especies naturales usadas como fármacos respecto de los productos de síntesis en regiones donde la tradición milenaria de los aborígenes ha convertido a los vegetales en recursos muy apreciados para una medicina natural utilizada, principalmente, por aquellos que no pueden acceder a las tratamientos de la medicina clásica. La creciente conciencia sobre la necesidad de preservar el patrimonio vegetal natural frente a la pérdida de especies por la acción depredadora del hombre, ha reavivado el interés en el conocimiento y estudio de las especies nativas. El objetivo del trabajo fue determinar en Morrenia odorata (Hook & Am.) Lindl., la presencia de sustancias farmacológicamente activas o que pudieran resultar tóxicas para el consumo humano. También se investigó la composición físico-química de los frutos mediante la aplicación de reacciones específicas. Los resultados de los ensayos de toxicidad en M. odorata muestran que la misma es apta para el consumo humano con fines terapéuticos. en medicina tradicional, preparada como decocción e infusión de raíz, hojas y frutos. Del análisis del contenido nutricional de los frutos, se deduce que M. odorata contiene cantidades más elevadas de los componentes mayoritarios y un mayor aporte energético respecto a otros frutos convencionales. Estos resultados permitirán la difusión del consumo de los frutos de esta especie como una alternativa en la alimentación humana.

Study on the protection of Asclepiadaceae against free radical injury / 中国病理生理杂志

AIM:To study the protection and mechanism of Asclepiadaceae against the damage of neuron by free radical. METHODS:The model of ischemia and damaged neuron induced by H 2O 2 was made respectively. The protection of Asclepiadaceae was observed with the measurement of contents of MDA in brain and cultured neuron, transudatory rate of LDH, breaking rate of DNA and clearance rate of?OH in cultured neuron. RESULTS:Asclepiadaceae decreased the raising of MDA in brain induced by ischemia. The raising of transudatory rate of LDH,breaking rate of DNA and content of MDA inducing by H 2O 2 in cultured neuron were also observed. The clearance rate of?OH in cultured neuron increased as the contents of Asclepiadaceae raised. CONCLUSION: The mechanism of Asclepiadaceae protecting the neuron is related to its ability to clean up free radical.