Cytotoxicity and the bioconversion strategy of Aristolochia spp

Aristolochia plants are notable from an ethnopharmacological viewpoint, but the relevance of these species for medicinal purposes has been debated because of their inherent toxicity. The convergence of these contrasting realities can be readily achieved using bioconversion methods, which have been shown to be useful tools for numerous applications, including the detoxification of biomass. In this context, methanolic extracts of leaves from Aristolochia triangularis and Aristolochia gibertii, as well as the feces of Battus polydamas larvae fed with leaves from these plants, were prepared, and their cytotoxic activities were evaluated on a human fibroblast cell line (GM07492). The leaf extracts were found to be cytotoxic, leading to reductions of 42.1 and 33.8% on cell viability, respectively, while the fecal extracts were considered inactive. In addition to evidencing the cytotoxicity of A. triangularis and A. gibertii, these findings demonstrated a potential bioconversion strategy for obtaining aristolochiaceous extracts with reduced toxicity using the larvae of a specialist phytophagous insect, thus renewing expectations in relation to the pharmacological importance of Aristolochia spp. The results were also ecologically relevant, as B. polydamas larvae were found to be able to detoxify compounds from host plants.(AU)

Simple and Rapid Differentiation of Mutong of Aristolochiaceae by Direct Spray Mass Spectrometry / 分析化学

Direct spray mass spectrometry was used to simply and rapidly differentiate Mutong of Aristolochiaceae from other two kinds of Mutong medicinal materials (Lardizabalaceae and Ranunculacea) by analyzing the chemical profile of Mutong of Aristolochiaceae.A novel method for determination of magnoflorine content in Mutong of Aristolochiaceae was established.The results showed that Mutong of Aristolochiaceae could be identified according to the symbolic component, magnoflorine.Under positive ion mode, semi-quantitative result based on the signal strength ratio of magnoflorine and nuciferin was obtained by choosing nuciferin as an internal standard.The method showed good linear coefficient in the concentration range of 0.50-20.00 mg/L of magnoflorine.The limit of detection was 0.1 mg/L.The method was simple and fast, and could be used for direct and rapid in-situ analysis and identification of Mutong of Aristolochiaceae from other closely related Mutong herbal species without sample pre-treatment.The results were important for the quality control of Mutong herbal medicine.

Antidiarrhoeal activity of Aristolochia argentina Gris. (Aristolochiaceae) in rodents.

The roots of Aristolochia argentina are used in folk medicine for the treatment of colitis, diarrhoea and hemorrhoids. In this study, based on ethnobotanical lead, we evaluated the antidiarrhoeal activity of Aristolochia argentina lyophilized aqueous extract (AALE) in rats and mice using various models. The castor oil and magnesium sulphate induced diarrhoea, the small intestinal transit in mice and the intestinal fluid accumulation were used in this study. At the doses of 62.5, 125 and 250 mg/kg p.o., the AALE showed significant antidiarrhoeal activity in both models. The AALE significantly reduced the intestinal fluid accumulation in the castor oil induced enteropooling. AALE delays small intestinal transit possibly, at least in part, involving opioid and α2-adrenergic receptors. The phytochemical analysis revealed the presence of carbohydrates, flavonoids, tannins, saponins and anthraquinones. The results suggest that AALE showed antidiarrhoeal activity by inhibiting intestinal motility and enteropooling property, justify its use in traditional medicine.

Proifling on Invention Patents ofXi-Xinin China / 世界科学技术-中医药现代化

Xi-Xin, a commonly used traditional Chinese herbal medicine, was regarded as one of the top grade materia medica since ancient time. The main medical parts were the root and stem. TheChinese Pharmacopeiaregulated the types ofAsarum heterotropoidesvar. mandshuricum,A. sieboldiivar. seoulense, andA. sieboldii. Analysis onXi-Xin related invention and patent information was an important aspect to understand the patent application condition and development tendency. It will also promote the research, development and industrialization of this medicine and promote its scientific result transformation. The Kongfz Patent Search Engine was used in the patent searching. Python and R languages were used in the collection, screening, classification, statistics, digging and analysis. In-depth analysis was made on the domesticXi-Xin related medical invention, patent application condition and development tendency. It provided references for the development ofXi-Xin. The results showed that an ever-increasing number of patents onXi-Xin were available. TheXi-Xin related inventions and patent applications were mainly from China, Republic of Korea, Japan, Russia, the U.S.A., etc. The main clinical applications were common cold, cough, headache, rheumatism and arthralgia, antibiosis, antiphlogosis, and cardiovascular diseases.Xi-Xin was mostly combined with herbal medicine with the effects of invigorating blood circulation and removing blood stasis, relieving exterior syndrome, and tonifying deficiency.

In vitro Regeneration of Plantlets from Leaf and Nodal explants of Aristolochia indica L.- An Important Threatened Medicinal Plant

Objective: An efficient reproducible protocol has been developed for in vitro regeneration of plantlets from leaf and nodal explants of Aristolochia indica L. Methods: Wild grown plants Aristolochia indica L. were collected and grown in the departmental garden. Leaf and nodal segments (0.5-1.0 cm) from young healthy plants were first washed thoroughly under running tap water for 15 - 20 minutes and then treated with liquid detergent [5% (v/v) Tween-20] for 5-10 minutes. Later these explants were washed with double-distilled water for 5 minutes. Subsequently, explants were immersed in 70% (v/v) ethanol for 2 - 3 minutes and washed with sterile glass double distilled water for 2-3 times. Eventually, the explants were treated with an aqueous solution of 0.1% (w/v) HgCl2 for 1 - 2 minutes and rinsed for two-to-three times in sterile ddH2O to remove all traces of HgCl2. The sterilized explants were inoculated aseptically onto solid basal Murashige and Skoog’s medium with different concentrations and combinations of BAP and NAA for in vitro regeneration of plants. Results: Both leaf and nodal explants cultured on MS medium supplemented with 0.8 mg/L BAP developed into mass of callus. These calli were subcultured for the induction of shoots and roots. Shoots were induced from both calli on MS medium supplemented with 0.8 mg/L BAP+0.5 mg/L NAA. Roots were induced from in vitro shoots on MS medium supplemented with 0.8 mg/L NAA for 4 weeks. Nodal explants were more regenerative with 95 % response compared to leaf explants with 85%. Finally, these in vitro regenerated plantlets were hardened, acclimatised and successfully transferred to the field. Conclusions: The present protocol for in vitro regeneration of Aristolochia indica L. can be used to make this plant available throughout the year for traditional healers, pharmaceutical usages, germplasm conservation, commercial cultivation, and also for the production of secondary metabolites.

Germinação de sementes de Aristolochia esperanzae O. Kuntze em diferentes temperaturas e condições de luminosidade / Germination of Aristolochia esperanzae O. Kuntze seeds under different temperatures and light conditions

O objetivo deste estudo foi verificar a influência da temperatura e da luz, em interação com a temperatura e de forma isolada, bem como, determinar a temperatura mais adequada para a condução do teste de germinação de sementes de Aristolochia esperanzae O. Kuntze (cipó mil-homens). Foram realizados três ensaios. No primeiro, as temperaturas utilizadas foram 15, 20, 25, 30, 35 e 40ºC. No segundo, foi avaliada a germinação das sementes nas temperaturas de 25, 30 e 35ºC, na presença e ausência de luz e no terceiro ensaio, foram testadas quatro condições de luminosidade, de forma isolada: (a) luz branca; (b) luz vermelha; (c) vermelho-distante e (d) ausência de luz, na temperatura de 25ºC. Em ambos os ensaios, foram avaliados, durante 30 dias, as porcentagens de sementes germinadas, de plântulas normais e de sementes não germinadas, e os tempos médios de sementes germinadas e de plântulas normais. Nas temperaturas de 15ºC e de 40ºC, não ocorreram germinação de sementes de A. esperanzae. O melhor resultado para sementes germinadas e para formação de plântulas normais (96 por cento) foi obtido na temperatura de 30ºC. A presença de luz e as temperaturas de 25ºC e 30ºC favoreceram a germinação de sementes e a formação de plântulas normais. As sementes de Aristolochia esperanzae tiveram sua germinação inibida na ausência de luz, caracterizando-as como fotoblásticas positivas.

The aim of this study was to verify the influence of temperature and light, interacting or separately, as well as to establish the most suitable temperature to conduct the germination test of Aristolochia esperanzae O. Kuntze ("cipó mil-homens") seeds. Three assays were performed. In the first one, temperatures were 15, 20, 25, 30, 35 and 40ºC. In the second assay, seed germination was evaluated at 25, 30 and 35ºC, in the presence and absence of light. In the third assay, four light conditions were tested separately: (a) white light; (b) red light; (c) far-red light; and (d) no light, at 25ºC. In all assays, the percentages of germinated seeds, normal seedlings and non-germinated seeds, besides the mean times of germinated seeds and normal seedlings, were evaluated during 30 days. At 15ºC and 40ºC, there was no A. esperanzae seed germination. The best result for germinated seeds and normal seedlings (96 percent) was obtained at 30ºC. The presence of light and the temperatures 25ºC and 30ºC favored seed germination and normal seedling formation. Aristolochia esperanzae seeds had their germination inhibited in the absence of light, which characterizes them as positive photoblastic.

Aristolochiaceae endémicas del Perú

La familia Aristolochiaceae es reconocida en el Perú por presentar un género, Aristolochia, y alrededor de 40 especies (Brako & Zarucchi, 1993), todas bejucos y lianas. En este trabajo reconocemos siete endemismos. La mayoría de estos taxones endémicos ocupan las regiones Bosques Muy Húmedos Premontanos y Bosques Húmedos Amazónicos, con dos especies ocupando la región Bosques Secos, entre los 200 y 2000 m de altitud. Solamente una especie endémica se encuentra representada dentro del Sistema Nacional de Áreas Naturales Protegidas por el Estado.

The Aristolochiaceae are represented in Peru by 40 species in the genus Aristolochia (Brako & Zarucchi, 1993), all of them vines and lianas. Here we recognize seven endemic species. Most of these endemic taxa are found in the Very Humid Premontane and Humid Lowland Amazonian Forest regions, between 200 and 2000 m elevation, while two species are found in the Dry Forest region. Only one endemic species has been found to date in protected areas.

Estudo fitoquímico das partes aéreas de Aristolochia birostris Ducht. (Aristolochiaceae) / Phytochemical study of the aerial parts of Aristolochia birostris Ducht. (Aristolochiaceae)

A família Aristolochiaceae apresenta entre seus constituintes terpenóides, alcalóides e lignóides. Papo-de-peru, jarrinha, mil-homens e capivara são nomes populares da espécie Aristolochia birostris, a qual é utilizada na medicina popular como antiofídica, sudorífica, anticatarral e abortiva. Um estudo fitoquímico anterior com as partes aéreas desta espécie reportou o isolamento de quatro terpenóides, um lignóide e uma antraquinona. Dando continuidade a esse estudo, este trabalho relata a identificação de 4 lignóides, sendo dois do tipo ariloxiarilpropânico (4-metilenodioxi-5,3',5' -trimetoxi-1'-alil-8.O.4'-neolignana e 3,4,5,3',5'-pentametoxi-1'-alil-8.O.4'-neolignana), um do tipo tetraidrofurano (grandisina) e uma lignana dibenzilbutirolactônica [(-)-hinoquinina], além de vanilina. As estruturas foram estabelecidas através de métodos espectroscópicos de infravermelho e ressonância magnética nuclear uni e bidimensionais. Os lignóides foram avaliados quanto a atividade antimicrobiana frente a algumas cepas de bactérias, fungos e leveduras, entretanto foram inativos.

The family Aristolochiaceae presents among its chemical constituents terpenoids, alkaloids and lignoids. Aristolochia birostris is a species of this family popularly known as "papo-de-peru", "jarrinha", "mil-homens" and "capivara", and it is used in the folk medicine to treat snake bites, as a sudorific, expectorant and abortive. A previous phytochemical study of the aerial parts of this plant reported the isolation of four terpenoids, lignoids and antraquinone. Continuing this study it is now reported the isolation of four lignoids, two of the type aryloxyarylpropane (4-methylenedioxy-5,3',5'-trimethoxy-1'-alyl-8.O.4' -neolignan and 3,4,5,3',5'-pentamethoxy-1'-alyl-8.O.4'-neolignan), one tetrahydrofuran (grandisin) and a dibenzylbutyrolactone lignans [(-)-hinokinin], and also vanillin. The chemical structures of the isolated compounds were established based on spectral data, mainly IR and NMR including 1D and 2D experiments. The lignoids were evaluated for their activity against some bacteria (Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis and Escherichia coli), fungi (Aspergilus flavus and Microsporum canis) and yeast (Candida albicans, Candida tropicalis and Criptococcus neoformans), but they were inactive.