A comprehensive review of Pfaffia glomerata botany, ethnopharmacology, phytochemistry, biological activities, and biotechnology.

ETHNOPHARMACOLOGICAL RELEVANCE: Pfaffia glomerata (Spreng.) Pedersen, Amaranthaceae, is found in South America, mainly in Brazil, where it is considered a species of great medicinal interest owing to its popular use as a tonic, aphrodisiac, anti-inflammatory, and analgesic. These properties can be attributed to the presence of the phytosteroid, 20-Hydroxyecdysone (ß-ecdysone), the main compound found in its roots. AIM OF THE REVIEW: This review aims to provide information about the botanical characteristics, ethnomedicinal uses, the phytochemistry, the biological activities, and the biotechnology of P. glomerata, an important species to local communities and groups researching medicinal plants of South America. MATERIALS AND METHODS: The information available on P. glomerata was collected from scientific databases (ScienceDirect, PubMed/MEDLINE, SciELO, and Scopus) until June 7, 2023, using the search terms "Pfaffia glomerata", "Pfaffia glomerata (Spreng.) Pedersen", and "Brazilian ginseng". The review includes studies that evaluated the botanical, ethnopharmacological, and phytochemical aspects, biological properties, nutraceutical uses, and the application of biotechnology for improving the biosynthesis of metabolites of interest. RESULTS: A total of 207 studies were identified, with 81 articles read in full. Seventy-six studies were included for qualitative synthesis. Overall, 40 compounds belonging to different classes are presented in this review, including ecdysteroids, triterpenes, saponins, flavonoids, anthraquinones, tannins, coumarins, alkaloids, and polysaccharides. Among them, flavonoids, anthraquinones, tannins, coumarins, and alkaloids were only putatively identified. ß-Ecdysone, triterpenes, saponins, and polysaccharides are the chemical components most frequently identified and isolated from P. glomerata and possibly responsible for ethnopharmacological use and the biological activities of this species, with important in vitro and in vivo activities, such as anti-inflammatory, antidepressant, aphrodisiac, analgesic, gastroprotective, antioxidant, and prebiotic. CONCLUSIONS: This review summarizes discussions about the P. glomerata species, highlighting its ethnopharmacological, chemical, biotechnological, and nutraceutical importance. New scientific studies on this species are encouraged in the search for new therapeutic molecules with pharmaceutical potential and nutraceutical applications.

Extract of Pfaffia glomerata Ameliorates Paroxetine-Induced Sexual Dysfunction in Male Mice and the Characterization of Its Phytoconstituents by UPLC-MS.

Pfaffia glomerata extract (PGE) has a variety of biological activities. However, its ameliorative effect on and exact working mechanism in male sexual dysfunction are still poorly understood. This study aims to evaluate the ameliorative effect of PGE on paroxetine (PRX)-induced sexual dysfunction in male mice and uses molecular docking technology to investigate its underlying mechanism. In this work, PRX-induced sexual dysfunction was caused and PGE was gavaged in mice for 28 days. The results show that PGE significantly improved the sexual performance of mice and reduced the damage to testicular tissues. Further studies showed that PGE restored serum sex hormones to normal levels and increased nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) levels as well as nitric oxide synthase (NOS) activity in penile tissues, while also decreasing phosphodiesterase-5 (PDE-5) activity, thereby maintaining normal penile erection in mice. In addition, PGE improved the activities of enzymes (LDH, ACP, and ALP) related to energy metabolism in the testis and significantly increased sperm count and viability in mice. Furthermore, the molecular docking results show that all eight compounds in PGE could form a stable complex with PDE-5 and inhibit the activity of PDE-5. In conclusion, PGE had an ameliorative effect on PRX-induced sexual dysfunction, suggesting that PGE has a potential protective effect on male sexual health.

β-ecdysone content and antioxidant capacity in different organs of Brazilian ginseng / Teor de β-ecdisona e capacidade antioxidante em diferentes órgãos de ginseng brasileiro

ABSTRACT: Plants that contain antioxidant compounds have attracted increasing interest for their vital role in the attenuation of oxidative damage caused by free radicals and in the treatment of various diseases. The present study investigated the β-ecdysone content and the antioxidant activity of Brazilian ginseng (Pfaffia glomerata) extracts obtained from inflorescences, stems, and roots. The P. glomerata extracts were tested for antioxidant activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method, β-carotene bleaching test, and phosphomolybdenum method. The β-ecdysone content of P. glomerata extracts was measured by high-performance liquid chromatography (HPLC). The P. glomerata inflorescences showed the strongest DPPH radical scavenging activity and the strongest antioxidant activity in the β-carotene bleaching assay and phosphomolybdenum test. The roots showed the lowest antioxidant capacity in all of the assays. The concentration of β-ecdysone in the plant organs followed the following decreasing order: inflorescences > stems > roots. The present study showed that P. glomerata inflorescence extract had high antioxidant capacity that could be attributed to the presence of β-ecdysone.

RESUMO: Plantas que contêm compostos antioxidantes têm atraído interesse crescente por seu papel fundamental na atenuação de danos oxidativos causados pelos radicais livres e no tratamento de várias doenças. O presente estudo investigou o conteúdo de β-ecdysone e a atividade antioxidante de extratos de ginseng brasileiro (Pfaffia glomerata) obtidos a partir das inflorescências, caules e raízes. Os extratos de Pfaffia glomerata foram testados para atividade antioxidante usando o método sequestrante do radical 2,2-difenil-1-picrilhidrazil (DPPH), sistema modelo β-caroteno-linoleato e método de fosfomolibdênio. O conteúdo de β-ecdisona dos extratos de P. glomerata foi medido por cromatografia líquida de alta eficência (CLAE). As inflorescências de P. glomerata mostraram a maior atividade sequestrante de radical DPPH e a maior atividade antioxidante no ensaio β-caroteno-linoleato e no teste de fosfomolibdênio. As raízes mostraram a menor capacidade antioxidante em todos os ensaios. A concentração de β-ecdisona nos órgãos da planta seguiu a seguinte ordem decrescente: inflorescências > caules > raízes. Os resultados indicaram uma correlação positiva entre conteúdo de β-ecdisona e atividade sequestrante de radical DPPH. O presente estudo mostrou que o extrato das inflorescências de P. glomerata teve alta atividade antioxidante que poderia ser atribuída à presença de β-ecdisona.

Evaluation of the testis function of mice exposed in utero and during lactation to Pfaffia glomerata (Brazilian ginseng).

Pfaffia glomerata (Spreng.) Pedersen, popularly known as "Brazilian ginseng," is used as medicinal plant in Brazil to treat inflammatory diseases in general. Previous studies showed that its extract increases the nitric oxide (NO) levels. Knowing that NO downregulates steroidogenesis and that alterations in the action/production of androgens during perinatal life could alter testis development, the present studies sought to investigate the reproductive toxicity of Pfaffia glomerata on male mice exposed to hydroalcoholic extract in utero and during lactation. The present study shows that P. glomerata extract does not alter body weight, tubular diameter and testis function in male mice. Although a reduction in the testis weight was observed in the animals that received the highest dose directly in early post-natal life, our findings show clearly that P. glomerata may not act as an endocrine disruptor, and it is not an "antiandrogenic" compound that could lead to testicular dysgenesis syndrome.

De novo assembly and transcriptome of Pfaffia glomerata uncovers the role of photoautotrophy and the P450 family genes in 20-hydroxyecdysone production.

Pfaffia glomerata is a medically important species because it produces the phytoecdysteroid 20-hydroxyecdysone (20-E). However, there has been no ready-to-use transcriptome data available in the literature for this plant. Here, we present de novo transcriptome sequencing of RNA from P. glomerata in order to investigate the 20-E production as well as to understand the biochemical pathway of secondary metabolites in this non-model species. We then analyze the effect of photoautotrophy on the production of 20-E genes phylogenetically identified followed by expression analysis. For this, total messenger RNA (mRNA) from leaves, stems, roots, and flowers was used to construct indexed mRNA libraries. Based on the similarity searches against plant non-redundant protein database, gene ontology, and eukaryotic orthologous groups, 164,439 transcripts were annotated. In addition, the effect of photoautotrophy in two genes putatively involved in the 20-E synthesis pathway was analyzed. The Phantom gene (CYP76C), a precursor of the route, showed increased expression in P. glomerata plants cultured under photoautotrophic conditions. This was accompanied by increased production of this metabolite indicating a putative involvement in 20-E synthesis. This work reveals that several genes in the P. glomerata transcriptome are related to secondary metabolism and stresses, that genes of the P450 family participate in the 20-E biosynthesis route, and that plants cultured under photoautotrophic conditions promote an upregulated Phantom gene and enhance the productivity of 20-E. The data will be used for future investigations of the 20-E synthesis pathway in P. glomerata while offering a better understanding of the metabolism of the species.

Comparison of oleanolic acid content in Pfaffia glomerata from different habitats of Guangxi / 国际药学研究杂志

Objective: To establish a method for the determination of oleanolic acid(OA)in Pfaffia glomerata to compare the contents of OA in different parts of P. glomerata from Guangxi, China, with different growth years and different habitats. Methods: HPLC method was adopted. The determination was performed on Phenomenex Luna C18(250 mm×4.6 mm, 5 μm)column with mobile phase consisting of methanol-water-glacial acetic acid-triethylamine(285:15: 0.2:0.1, V/V) at the flow rate of 1.0 ml/min. The column temperature was set at 30℃. The detection wavelength was set at 210 nm, and the injection volume was 20 μl. The content of OA in P. glomerata was determined by the established method, and the extraction process of OA was optimized by the orthogonal test. Results: The linear range of OA was 0.0206-2.060 mg/ml(r2=1.0000). RSDs for the precision, repeatability and stability tests were all lower than 2%(n=6 or n=7). The average recovery of OA was 100.89%(RSD=1.69%, n=9). The optimum extraction conditions of OA were as follows: 20-fold ethanol(80%, V/V), extracting for three times, refluxing 1.5 hour each time, and processing acid hydrolysis with 3.0% sulfuric acid(g/g)for 1 hour. Under these conditions, the OA had the highest extraction efficiency. The contents of OA in reed head, root, old stem, tender stem, leaf and flower of P. glomerata were 4.87, 4.61, 2.67, 0.99, 0.24 and 1.13 mg/g, respectively. The average contents of OA in the roots of P. glomerata aged 1, 2, 3, 4 and 5 years in Guangxi were 3.08, 4.07, 4.71, 4.62 and 4.46 mg/g, respectively. Conclusion: The established extraction process and detection method is suitable for the extraction and content determination of OA in P. glomerata. Although OA is distributed in all parts of P. glomerata, the contents significantly vary in different parts. The content of OA is highest in reed head and lowest in leaves. The OA content in P. glomerata becomes stable after 3 years of growth in Guangxi.

Optimization of the extraction and enrichment process and evatuation of the free radical scavenging activity of triterpene sapogenins from Pfaffia glomerata / 国际药学研究杂志

Objective: To establish a macroporous resin enrichment process for total triterpene sapogenins from Pfaffia glomerata,and evaluate the free radical scavenging activity of the total triterpene sapogenins. Methods: The transfer rate of triterpene sapogenins was used as the index. Static adsorption-desorption method was used to select the best macroporous resin,dynamic adsorption-desorption method was used to determine the enrichment process parame- ters,and orthogonal test was used to optimize the enrichment process. The 1,1- diphenyl- 2- trinitrophenylhydrazine (DPPH)and hydroxyl free radical scavenging tests were used to evaluate the free radical scavenging capacity of the triter- pene sapogenins. Results: The AB-8 type macroporous resin had the best enrichment effect on the triterpene sapoge-nins. The conditions for the optimum enrichment process were as follows:the concentration of triterpene sapogenins in the sample solution for resin column chromatography was 3.5 mg/ml,the ratio of diameter to height for the bed of resin column was 1:7,the amount of the raw herbs subjected to the resin column was 0.11 g/ml(herbs/resin),the flow rate for subjecting sample solution to resin column was 1 BV/h,the eluent was 95% aqueous ethanol,the eluting flow rate was 2BV/h,and the eluent volume for elution was 7 BV. After optimization,total triterpene sapogenins reached 65% in the samples prepared by the optimized process,and the transfer rate of total triterpene sapogenins reached 80.23% in the pre- paring process. The total triterpene sapogenin sample could scavenge the DPPH and hydroxyl free radicals,and the scav- enging rate for the DPPH and hydroxyl radicals reached 82.42% and 73.43% at the 1.0 mg/ml,respectively. Conclusion: The AB- 8 macroporous resin can be used for the enrichment and purification of triterpene sapogenins from P. glomerata. The optimized enrichment process was stable and feasible,and the obtained triterpene sapogenins showed a good free radical scavenging activity.

Selenium and silicon reduce cadmium uptake and mitigate cadmium toxicity in Pfaffia glomerata (Spreng.) Pedersen plants by activation antioxidant enzyme system.

Cadmium (Cd) is toxic to plants and animals, making it necessary to develop strategies that seek to reduce its introduction into food chains. Thus, the aim of this study was to investigate whether silicon (Si) and selenium (Se) reduce Cd concentrations in Pfaffia glomerata medicinal plant and attenuate the oxidative stress promoted by this metal. These plants were cultivated in hydroponics under the following treatments: control (nutrient solution), 2.5 µM Se, 2.5 mM Si, 50 µM Cd, 50 µM Cd + 2.5 µM Se, 50 µM Cd + 2.5 mM Si. After 14 days of exposure to treatments, leaves and roots were collected for the determination of dry weight of shoot and roots, Cd concentrations, chlorophyll and carotenoids content, and biochemical parameters (lipid peroxidation and guaiacol peroxidase and superoxide dismutase activities). The data were submitted to analysis of variance and means were compared with Scott-Knott test at 5% error probability. Roots of P. glomerata plants showed a significant reduction on dry weight accumulation when exposed to Cd. However, both Se and Si promoted a significant reduction of deleterious effects of Cd. The Cd concentrations in the tissues were reduced in the presence of Se or Si. Plants treated with Cd together with Se or Si presented higher pigment content than those with only Cd, thus showing a reduction in the negative effects caused by this element. In the treatments in which Se and Si were added in the growth medium together with Cd, an activation of superoxide dismutase and guaiacol peroxidase enzymes was observed in the roots and shoot, which may have contributed to lower lipid peroxidation. Thus, Se and Si reduce Cd concentrations and have potential to ameliorate Cd toxicity in P. glomerata plants, which can be used to increase productivity and quality of medicinal plants.

Three new noroleanane-type triterpenes from the roots of Pfaffia glomerata.

The Amaranthaceae plant, Pfaffia glomerata, which is so-called as Brazil ginseng, is widely distributed in South American countries. Three new noroleanane-type triterpenes and four known oleanane-type triterpenes were isolated from the roots of P. glomerata. Their structures were determined by spectroscopic methods including 1D and 2D NMR experiments. Their effects on melanogenesis were also reported.

Chemical constituents from ethyl acetate extracts of domestic Pfaffia glomerata / 中草药

Objective: To study the chemical constituents of ethyl acetate extract from domestic Pfaffia glomerata. Methods: The compounds were isolated and purified by silica gel, reversed-phase, Sephadex LH-20 column chromatography, and their structures were elucidated by spectroscopic analysis as well as chemical methods. Results: Twenty compounds were identified as 1-undecanol (1), oleic acid (2), β-sitosterol (3), 2-hydroxy-4,6-dimethoxyacetophenone (4), oleanolic acid (5), pfaffianol A (6), benzene-1,4-diol (7), vanillic acid (8), iresinone (9), ethylcaffeate (10), oleanoicaeid-28-O-β-D-glucopyranosyl ester (11), allantoin (12), ajugasterone-C (13), β-ecdysterone (14), iresinoside (15), adenine (16), oleanolic acid 3-O-β-D-glucuronopyranoside (17), ficusoside B (18), pfaffiaglycoside B (19), and β-D-Glucopyranosyl-3-(O-β-D-glucopyranosyloxy)-oleanolate (20). Conclusion: Compounds 7-10, 15, 17, 18, and 20 are isolated from this plant for the first time.

In vitro Activities of Pfaffia glomerata Root Extract, Its Hydrolyzed Fractions and Pfaffic Acid Against Trypanosoma cruzi Trypomastigotes.

This article reports on the in vitro activity of the hydroalcoholic extract of Pfaffia glomerata roots, its hydrolyzed fractions, and pfaffic acid against Trypanosoma cruzi. The hydroalcoholic extract obtained from dried, milled P. glomerata roots was submitted to acid hydrolysis followed by partition with CHCl3 . The concentrated CHCl3 fraction was suspended in MeOH/H2 O and partitioned with hexane (F1), CHCl3 (F2), and AcOEt (F3), in this sequence. The trypanocidal activity of the hydrolyzed extract and its fractions was evaluated in vitro. The hydroalcoholic extract displayed low activity, but fraction F1 was active against trypomastigotes of the Y strain of T. cruzi, with IC50 = 47.89 µg/ml. The steroids campesterol (7.7%), stigmasterol (18.7%), ß-sitosterol (16.8%), Δ7 -stigmastenol (4.6%), and Δ7 -spinasterol (7.5%) were the major constituents of F1, along with fatty acid esters (7.6%) and eight aliphatic hydrocarbons (30.1%). Fractions F2 and F3 exhibited moderate activity, and pfaffic acid, one of the main chemical constituents of these fractions, displayed IC50 = 44.78 µm (21.06 µg/ml). On the other hand, the hydroalcoholic extract of P. glomerata roots, which is rich in pfaffosides, was inactive. Therefore, the main aglycone of pfaffosides, pfaffic acid, is much more active against trypomastigotes of the Y strain of T. cruzi than its corresponding glycosides and should be further investigated.

The phytoecdysteroid ß-ecdysone is genotoxic in Rodent Bone Marrow Micronuclei and Allium cepa L. Assays.

ETHNAOPHARMACOLOGIAL RELEVANCE: In South America, the ß-ecdysone ecdysteroid has been found in species of the genus Pfaffia Mart. Due to the similar morphology of its roots to the Panax ginseng C. A. Mey. (Korean ginseng), some species of this genus has been known as Brazilian ginseng and have been used as tonic and aphrodisiac, as well as for the treatment of diabetes and rheumatism. AIM OF THE STUDY: Here we report a cytogenotoxic evaluation of ß-ecdysone (a natural ecdysteroid found in plants) in Rodent Bone Marrow Micronuclei and Allium cepa Assays. MATERIALS AND METHODS: Three ß-ecdysone (pure) concentrations (based in human therapeutic dosage) were used in the Micronucleus Assay. The animals were treated during two consecutive days. Micronucleated cells were counted in 2000 polychromatic erythrocytes per animal. For A. cepa L. Assay, one ß-ecdysone concentration was analyzed. The onions bulbs were exposed for 24h. RESULTS: The Micronucleus Assay showed genotoxic effects for all treatments, expressed by an increase of micronucleated cells. In A. cepa L. Assay, cell abnormalities associated to the malfunction/non-formation of mitotic spindle (aneugenic effect) and chromosomal bridges (clastogenic effect) were observed. CONCLUSIONS: The results indicate a cytogenotoxic activity of ß-ecdysone. Therefore, the popular use of Pfaffia and others species containing ß-ecdysone should be considered with caution.

Isolation and prebiotic activity of inulin-type fructan extracted from Pfaffia glomerata (Spreng) Pedersen roots.

Pfaffia glomerata (Amaranthaceae) is popularly known as "Brazilian ginseng." Previous studies have shown that fructose is the major carbohydrate component present in its roots. Inulin-type fructans, polymers of fructose, are the most widespread and researched prebiotics. Here, we isolated and chemically characterized inulin extracted from P. glomerata roots and investigated its potential prebiotic effect. Fructans were isolated and their structures were determined using colorimetric, chromatography, polarimetry, and spectroscopic analysis. The degree of polymerization (DP) was determined, and an in vitro prebiotic test was performed. The structure of inulin was confirmed by chromatography and spectroscopic analysis and through comparison with existing data. Representatives from the genera Lactobacillus and Bifidobacterium utilized inulin from P. glomerata, because growth was significantly stimulated, while this ability is strain specific. The results indicated that inulin extracted from P. glomerata roots represents a promising new source of inulin-type prebiotics.

Supplementation with Pfaffia glomerata (Sprengel) Pedersen does not affect androgenic-anabolic parameters in male rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Paffia spp (Amaranthacea) has a widespread use of in Brazil as a possible hormonal supplement and a substitute of Panax ginseng, although information on its reproductive effects is missing. AIM OF THE STUDY: To evaluated possible anabolic-androgenic or anti-androgenic effects of Pfaffia glomerata (PG) extract using intact eight-months-old male rats and pre-pubertal castrated rats. MATERIALS AND METHODS: Three different dose levels of PG (8.5, 30 and 85 mg/kg/day) were administered to eight-months-old rats for 28 days or to castrated males for 7 days (Hershberger assay). In the experiment with intact animals, 24h fecal samples were collected for quantification of fecal metabolites of androgens throughout treatment. At the end of the treatment period, animals were euthanized for evaluation of serum testosterone, reproductive organ weights, number of spermatids per testis, diameter of seminiferous tubules and cross-sectional area of soleus muscle fibers. In the Hershberber assay, androgenic or anti-androgenic effects were evaluated by the weights of androgen-dependent tissues: ventral prostate, seminal vesicle, glans penis and levator ani muscle/bulbocavernosus muscle. RESULTS: No effects were observed in the concentrations of fecal metabolites of androgens monitored during the treatment of intact eight-months-old rats. Moreover, at the end of treatment, no changes were seen in any of the investigated parameters. In the Hershberger assay, the PG extract did not induce androgenic or anti-androgenic effects at the dose levels tested. Significant effects were only observed in animals treated with testosterone and testosterone plus flutamide, which were used as positive controls for androgenicity and anti-androgenicity, respectively. CONCLUSIONS: At the dose levels tested, PG extract does not induce anabolic-androgenic or anti-androgenic effects in rats.

Quantification of β-ecdysone in differents parts of Pfaffia glomerata by HPLC

Pfaffia glomerata (Spreng.) Pedersen, Amaranthaceae, is widely distributed in Brazil. Roots are considered as the world's greatest supplier and β-ecdysone is the most important compound extracted from roots of Pfaffia glomerata. So, the aim this study was analyze the presence of β-ecdysone in the inflorescences and stems and compared with the content from roots of Pfaffia glomerata and determine the best extractive method of β-ecdysone this plant. The crude extracts were obtained by Soxhlet method, reflux, maceration, percolation and turbolyse. Compound extracts were quantified by High Performance Liquid Chromatography (HPLC). The analysis were carried out a Phenomenex Column C18, 5 µm, 250x4,6mm, maintened at 30 ºC, gradient system using as mobile phase a mixture of methanol and water, flow rate 1,0 mL and detection at 245 nm. Results showed Soxhlet method with ethanol:water (90:10 v/v) presented the higher concentration of β-ecdysone in P. glomerata and inflorescences showed higher amount of this active substance (3,06%), compared with stems (2,37%) and roots (1,63%), showing that the inflorescences and plant stems may also be used as a rich source of β-ecdysone.

Extração de ecdisterona em raízes de ginseng brasileiro / Extraction of ecdysterone from roots of Brazilian ginseng

Este estudo teve como objetivo otimizar a extração de ecdisterona em raízes de ginseng brasileiro. Primeiramente, para se avaliar a eficiência do solvente extrator, amostras de raízes dois acessos (BRA e JB-UFSM) de P. glomerata foram extraídas em Soxhlet com metanol e clorofórmio, separadamente, durante 4 horas. No segundo ensaio, com o intuito de se escolher o método extrator, a extração foi conduzida em Soxhlet e em ultrassom, utilizando metanol como solvente. Em P. tuberosa, as amostras foram extraídas com metanol, e a extração foi conduzida em Soxhlet e em banho ultrasônico. O conteúdo de ecdisterona foi determinado em Cromatógrafo Líquido de Alta Eficiência (CLAE). Em ambas as espécies, um maior conteúdo de ecdisterona foi detectado nas amostras extraídas com metanol e em Soxhlet. A metodologia proposta mostrou-se eficaz para a quantificação da ecdisterona a partir das raízes de P. glomerata e P. tuberosa, podendo ser aplicada no controle de qualidade de drogas vegetais e/ou fitoterápicos.

This study aimed at optimizing the extraction method from ecdysterone of Brazilian ginseng. Root samples of two accessions (BRA and JB-UFSM) of P. glomerata were extracted in a Soxhlet with methanol or chloroform for 4h. In the second trial, the extration was conduced in a Soxhlet or ultrasonic using metanol as a solvent. In P. tuberosa, the roots samples were extracted with methanol in a Soxhlet or in ultrasonic. The ecdysterone content was determinated using high efficiency liquid chromatography methods. In both studied species, the highest ecdisterone content was detected from samples extracted in a Soxhlet and using methanol as a solvent. This extration method has been successfully applied for determination of ecdysterone content from roots of Brazilian ginseng, and could be useful for the quality control of drugs and pharmaceutical formulations.

pH do meio de cultivo e crescimento de plântulas de ginseng brasileiro cultivadas in vitro / Culture medium pH and growth of brazilian ginseng in vitro cultured plantlets

O presente trabalho objetivou avaliar o efeito do pH do meio de cultivo sobre alguns parâmetros de crescimento da Pfaffia glomerata (Spreng.) Pedersen cultivada in vitro, bem como checar se o crescimento dos explantes altera o pH do meio ao longo do período de cultivo. Foram testados quatro tratamentos constituídos de distintos valores de pH (3,7; 5,0; 6,0 e 7,5) do meio de cultivo. O pH do meio de cultivo foi ajustado antes da inclusão do agar (6g L-1 - Merck) e da autoclavagem. Como fonte de explantes foram utilizadas segmentos nodais de plantas previamente estabelecidas in vitro em meio MS. Dos nove aos 15 dias após a inoculação (DAI) dos segmentos nodais, verificou-se maior número de raízes em pH 6,0 e o menor no pH 7,5. Aos 35 DAI, o comprimento da maior brotação e o número total de segmentos nodais por planta foram maiores em torno de pH 6,0. Aos 35 DAI, observou-se menor crescimento em biomassa de raízes em pH 3,7. Já a parte aérea apresentou menor biomassa em pH 7,5. Aos 35 DAI, a produção de matéria fresca e seca total da plântula foi maior em pH próximo a 6,0. Concluiu-se que valores de pH do meio de cultivo próximos a 6,0, ajustados antes da autoclavagem, são ideais para o crescimento da P. glomerata cultivada in vitro. Também se verificou que o crescimento da plântula modificou significativamente o pH do meio de cultivo.

The present research aimed to evaluate the effect of culture medium pH on some growth parameter of Pfaffia glomerata (Spreng.) Pedersen in vitro cultured plantlets, as well as to check whether the explant´s growth alters the culture medium pH. Four treatments consisted of different values (3.7; 5.0; 6.0 and 7.5) of culture medium pH were tested. The culture medium pH was adjusted prior to the addition of agar (6g L-1 - Merck) and autoclaving. Nodal segments from asseptic plants grown in MS medium were used as explants. From 9 to 15 days after inoculation (DAI) of nodal segments, the higher number of roots was obtained at pH 6.0, and the lower at pH 7.5. At 35 DAI, both length of the higher sprout and total number of nodal segments per plantlet were greater at about pH 6.0. At 35 DAI, roots biomass was lower at pH 3.7. On the other hand, shoots biomass was lower at pH 7.5. Fresh and dry matter of the whole plantlet was greater at pH around 6.0. In conclusion, values of culture medium pH near to 6.0, adjusted prior to autoclaving, are ideal for the growth of P. glomerata in vitro cultured plantlets. Moreover, the in vitro growth of plantlet modified significantly the culture medium pH.

Effects of Pfaffia glomerata (Spreng) pedersen aqueous extract on healing acetic acid-induced ulcers

The present study was carried out to evaluate the acute toxicity and the effect of the aqueous extract of the roots from Pfaffia glomerata (Spreng) Pedersen (Amaranthaceae) (AEP) on the prevention of acetic acid-induced ulcer and on the healing process of previously induced ulcers. The acute toxicity was evaluated in Swiss mice after oral administration of a single dose and the chronic gastric ulcer was induced with local application of acetic acid. The results showed that the LD50 of the extract was 684.6 mg.kg-1 for the intraperitoneal administration and higher than 10 mg.kg-1by the oral route. The administration of the AEP did not prevent ulcers formation. However, the AEP increased of the healing process of previously induced ulcers. The results suggest that AEP chronically administered promote an increase of tissue healing, after the damage induced by acetic acid and the extract seemed to be destituted of toxic effects in the mice by the oral route.

Pfaffia glomerata (Spreng) Pedersen (Amaranthaceae), uma planta conhecida popularmente como "Ginseng Brasileiro" e "paratudo", é utilizada para tratar distúrbios gástricos e como cicatrizante. Em estudos anteriores, foi demonstrado que o extrato aquoso bruto da P. glomerata (AEP) protegeu a mucosa gástrica contra úlceras induzidas por etanol e estresse e reduziu a secreção ácida gástrica basal e estimulada em ratos com ligadura de piloro. Além disso, a secreção gástrica de animais tratados com AEP apresentou níveis de nitrato e nitrito aumentados. O objetivo deste estudo foi avaliar se o AEP previne o desenvolvimento de úlceras induzidas por ácido acético e o efeito desse extrato no processo de cicatrização em úlceras previamente formadas. A administração do AEP em diferentes doses produziu efeitos tóxicos baixos e não preveniu a formação de úlceras, porém aumentou o processo de cicatrização em úlceras já existentes, como evidenciado no estudo histopatológico. Em conclusão, o AEP administrado cronicamente promove o aumento da cicatrização do tecido após a lesão induzida com o ácido acético.

Substituição de nitrato de potássio (PA) por salitre potássico no preparo de meio de cultura de tecidos vegetais esterilizado com hipoclorito de sódio / Potassium nitrate substitution for potassic saltpetre on preparation of plant tissue culture nutritive media esterilized with sodium hypochlorite

A aquisição de nitrato de potássio PA, para qualquer finalidade, depende de autorização do Ministério da Defesa, o que resulta em dificuldades na sua aquisição. Com a finalidade de estudar a possibilidade da substituição desse reagente por outro produto encontrado livremente no comércio, foi conduzido um experimento onde se testaram várias concentrações de salitre potássico em comparação com o nitrato de potássio PA, como componente dos sais de MS. Utilizou-se o produto granulado MURER® nas seguintes concentrações (g L-1): a) 7,0; b) 7,4; c) 7,8; d) 8,2. Foi adicionado um tratamento-controle, com o KNO3 PA, conforme aparece nos sais de MS (1,9 g L-1), sendo a espécie testada à fáfia, a partir de segmentos nodais. O meio de cultura constituiu-se dos sais de MS, vitaminas de White, 30 g L-1 sacarose, 2,0 mg L-1 ácido indol butírico, 2,0 mg L-1 ácido naftalenoacético, 2,0 mg L-1 benzilaminopurina, pH 6,0 + 0,1 e 1,5 g L-1 Phytagel® como gelificante. O número médio de ramos formados para todos os tratamentos foi inferior ao do tratamento-controle; mas, em compensação, o comprimento médio deles foi superior, também em todos os tratamentos, sendo que o comprimento total de ramos na menor concentração de salitre, atingiu 17,0 cm, enquanto no tratamento-controle foi de 12,6 cm. A média da biomassa seca foi de 68,3 mg para a menor concentração de salitre e de 50,0 mg para o tratamento-controle. A análise dos resultados indicou ser vantajosa a substituição do nitrato de potássio PA pelo salitre potássico.

The acquisition of potassium nitrate for any purpose, including preparation of nutritive media for micropropagation, depends on authorization from the Ministry of Defense, what turns out in difficulties in its acquisition. With the purpose of studying the possibility of replacing this component of the culture for another product found freely in the market, an experiment was carried out, where several concentrations of potassic saltpetre were tried, in comparison to the chemically pure potassium nitrate, as a component of the MS salts formulation. The commercial granulated product MURER® was used in the following (g L-1): a) 7,0; b) 7,4; c) 7,8; d) 8,2; and e) chemically pure potassium nitrate (control-treatment) as it appears in the MS salts, or 1,9 g L-1Fáfia was tested, starting from nodal segments. The basal nutrient medium comprised the MS salts, White vitamins, 30 g L-1 sucrose, 2.0 mg L-1 indol butyric acid, 2.0 mg L-1 naftaleneacetic acid, 2.0 mg L-1 benzilaminopurine, pH 6.0 + 0.1 and 1.5 g L-1 Phytagel® as the gelling agent. The number of branches formed in all concentrations of potassic saltpetre was lower than those ones in the control treatment; however, their lengths were higher. The mean length of branches in the treatment with the smallest potassic saltpetre concentration, reached 17.0 cm in length, while in the control-treatment it was 12,6 cm. The dry biomass weight reached 68.3 mg for the smallest potassic saltpetre concentration, but only 50,0 mg for the control treatment. The results indicated to be advantageous the substitution of the chemically pure potassium nitrate for the potassic saltpeter, when either shoot elongation or biomass production is desired, being the nutrient medium sterilized with sodium hipoclorite. However, when a faster multiplication rate is the main objective, the nutrient medium should be prepared with chemically pure potassium nitrate.

Hypnotic effect of ecdysterone isolated from Pfaffia glomerata (Spreng.) Pedersen / Efeito hipinótico de ecdisterona isolada de Pfaffia glomerata (Spreng.) Pedersen

Neste trabalho foi avaliado, em roedores, o efeito depressor das frações clorofórmio (CHCl3), acetato de etila (EtOAc) e n-butanol, obtidas das partes subterrâneas de Pfaffia glomerata, empregando-se o teste de tempo de sono barbitúrico como referência. Somente a fração lipofílica (CHCl3:EtOAc, 1:1, m/m) (i.p. 500 mg/kg; v.o. 1000 mg/kg) potenciou o tempo de sono induzido por pentobarbital. A ecdisterona foi isolada e identificada como constituinte majoritário (1,4 por cento m/m) desta fração, através de cromatografia líquida de alta eficiência e métodos espectroscópicos, respectivamente. Este composto potenciou o tempo de sono barbitúrico (100 mg/kg, i.p.; 400 mg/kg, v.o), sem causar hipotermia. Nestas mesmas doses, a ecdisterona não alterou a performance dos animais no rota-rod, esquiva inibitória e labirinto em cruz-elevado, além de não alterar o padrão de convulsões induzidas por pentilenotetrazol. Este perfil indica que esta substância, nestas doses, não apresenta perfil ansiolítico ou neurotóxico. Estes resultados indicam que a ecdisterona é o componente responsável pela ação hipnótica apresentada pela fração lipofílica obtida das partes subterrâneas de P. glomerata.

In this study the depressant effect of fractions from P. glomerata was initially evaluated using the mice barbiturate sleeping time test as reference. The fractions tested were the CHCl3, the EtOAc, the n-BuOH and the aqueous fraction obtained from P. glomerata subterraneous parts. Only the pretreatment with the lipophilic fraction (CHCl3: EtOAc, 1:1, w/w) increased the barbiturate sleeping time (i.p 500 mg/kg; v.o. 1000 mg/kg). Ecdysterone, the main substance isolated from this lipophilic fraction, was identified by spectroscopic methods and its content in the ethanol extract was determined as 1.4 percent (w/w) by HPLC. In order to investigate the hypothesis of ecdysterone displaying a depressant effect on nervous central system, an evaluation toward the hypnotic-sedative and anxiolytic effects of this drug was carried out. Ecdysterone 100 mg/kg, i.p, increased the barbiturate sleeping time without provoking hypothermia; when administered by oral route its minimal effective dose was 400 mg/kg. On the other hand, ecdysterone (100 mg/kg, i.p; 400 mg/kg, p.o) did not impair motor coordination and was ineffective on pentylenetetrazole-induced convulsion, elevated plus-maze and step-down inhibitory avoidance tests, indicating that at these doses the drug does not present an anxiolytic profile and does not cause manifest neurotoxic effects as well. In conclusion, the lipophilic fraction from P. glomerata presents a hypnotic effect being ecdysterone one of the compounds responsible for this CNS activity.