Hepatoprotective Effects of Four Brazilian Savanna Species on Acetaminophen-Induced Hepatotoxicity in HepG2 Cells.

We investigated four Cerrado plant species, i.e., Cheiloclinium cognatum (Miers) A.C.Sm, Guazuma ulmifolia Lam., Hancornia speciosa Gomes, and Hymenaea stigonocarpa Mart. ex Hayne, against acetaminophen toxicity using an in vitro assay with HepG2 cells. The activity against acetaminophen toxicity was evaluated using different protocols, i.e., pre-treatment, co-treatment, and post-treatment of the cells with acetaminophen and the plant extracts. HepG2 cell viability after treatment with acetaminophen was 39.61 ± 5.59% of viable cells. In the pre-treatment protocol, the extracts could perform protection with viability ranging from 50.02 ± 15.24% to 78.75 ± 5.61%, approaching the positive control silymarin with 75.83 ± 5.52%. In the post-treatment protocol, all extracts and silymarin failed to reverse the acetaminophen damage. In the co-treatment protocol, the extracts showed protection ranging from 50.92 ± 11.14% to 68.50 ± 9.75%, and silymarin showed 77.87 ± 4.26%, demonstrating that the aqueous extracts of the species also do not increase the toxic effect of acetaminophen. This protection observed in cell viability was accompanied by a decrease in ROS. The extracts' hepatoprotection can be related to antioxidant compounds, such as rutin and mangiferin, identified using HPLC-DAD and UPLC-MS/MS. The extracts were shown to protect HepG2 cells against future APAP toxicity and may be candidates for supplements that could be used to prevent liver damage. In the concomitant treatment using the extracts with APAP, it was demonstrated that the extracts do not present a synergistic toxicity effect, with no occurrence of potentiation of toxicity. The extracts showed considerable cytoprotective effects and important antioxidant characteristics.

Lycorine Alkaloid and Crinum americanum L. (Amaryllidaceae) Extracts Display Antifungal Activity on Clinically Relevant Candida Species.

Candida species are the main fungal agents causing infectious conditions in hospital patients. The development of new drugs with antifungal potential, increased efficacy, and reduced toxicity is essential to face the challenge of fungal resistance to standard treatments. The aim of this study is to evaluate the in vitro antifungal effects of two crude extracts of Crinum americanum L., a rich alkaloid fraction and lycorine alkaloid, on the Candida species. As such, we used a disk diffusion susceptibility test, determined the minimum inhibitory concentration (MIC), and characterized the components of the extracts using Electrospray Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (ESI FT-ICR MS). The extracts were found to have antifungal activity against various Candida species. The chemical characterization of the extracts indicated the presence of alkaloids such as lycorine and crinine. The Amaryllidaceae family has a promising antifungal potential. Furthermore, it was found that the alkaloid lycorine directly contributes to the effects that were observed for the extracts and fraction of C. americanum.

Seasonal Chemical Evaluation of Miconia chamissois Naudin from Brazilian Savanna.

Miconia chamissois Naudin is a species from the Cerrado, which is being increasingly researched for its therapeutic potential. The aim of this study was to obtain a standardized extract and to evaluate seasonal chemical variations. Seven batches of aqueous extracts from leaves were produced for the standardization. These extracts were evaluated for total solids, polyphenol (TPC) and flavonoid content (TFC), vitexin derivative content, antioxidant activity; thin-layer chromatography (TLC), and high-performance liquid chromatography (HPLC) profiles were generated. For the seasonal study, leaves were collected from five different periods (May 2017 to August 2018). The results were correlated with meteorological data (global radiation, temperature, and rainfall index). Using chromatographic and spectroscopic techniques, apigenin C-glycosides (vitexin/isovitexin) and derivatives, luteolin C-glycosides (orientin/isoorientin) and derivatives, a quercetin glycoside, miconioside B, matteucinol-7-O-ß-apiofuranosyl (1 → 6) -ß-glucopyranoside, and farrerol were identified. Quality parameters, including chemical marker quantification by HPLC, and biological activity, are described. In the extract standardization process, all the evaluated parameters showed low variability. The seasonality study revealed no significant correlations (p < 0.05) between TPC or TFC content and meteorological data. These results showed that it is possible to obtain extracts from M. chamissois at any time of the year without significant differences in composition.

Biodiversity of ß-Carboline Profile of Banisteriopsis caapi and Ayahuasca, a Plant and a Brew with Neuropharmacological Potential.

Ayahuasca is a psychoactive infusion with a large pharmacological application normally prepared with Banisteriopsis caapi, which contains the monoamine oxidase inhibitors ß-carbolines, and Psichotria virids, which contains the serotonin receptor agonist N,N dimethyltryptamine (DMT). The objectives of this study were to investigate the chemical profile of B. caapi and of ayahuasca collected in various Brazilian regions. In total, 176 plant lianas, of which 159 B. caapi and 33 ayahuasca samples were analyzed. Dried liana samples were powdered, extracted with methanol, diluted, and analyzed by LC-MS/MS. Ayahuasca samples were diluted and analyzed. Mean concentrations in B. caapi were 4.79 mg/g harmine, 0.451 mg/g harmaline, and 2.18 mg/g tetrahydroharmine (THH), with a high variability among the samples (RSD from 78.9 to 170%). Native B. caapi samples showed significantly higher harmine concentrations than cultivated ones, and samples from the Federal District/Goiás had higher THH content than those collected in the State of Acre. The other Malpighiaceae samples did not contain ß-carbolines, except for one D. pubipetala sample. Concentrations in ayahuasca samples ranged from 0.109 to 7.11 mg/mL harmine, 0.012 to 0.945 mg/mL harmaline, 0.09 to 3.05 mg/mL THH, and 0.10 to 3.12 mg/mL DMT. The analysis of paired ayahuasca/B. caapi confirmed that harmine is reduced to harmaline and to THH during the brew preparation. This is the largest study conducted with Malpighiaceae samples and showed a large variability in the main ß-carbolines present in B. caapi. This biodiversity is a challenge for standardization of the material used in ethnopharmacological studies of B. caapi and ayahuasca.

Triterpenes from Pouteria ramiflora (Mart.) Radlk. Leaves (Sapotaceae).

Pouteria ramiflora (Mart.) Radlk. (Sapotaceae) is a species used by inhabitants from the Cerrado for its edible fruits and medicinal value. Hexane crude extracts from leaves and fractions were evaluated for in vitro α-amylase inhibitory activity and antioxidant potential. The fraction with the highest α-amylase inhibitory activity was submitted to a phytochemical study. Three triterpenes were isolated, friedelin, epi-friedelanol, and taraxerol. This is the first report of these compounds isolated from P. ramiflora. Moreover, this is the first report of friedelin isolated from Pouteria sp. Epi-friedelanol was present in significant amounts, suggesting that this compound could be a candidate marker for this species.

Activity of crude extracts from Brazilian cerrado plants against clinically relevant Candida species.

BACKGROUND: Medicinal plants have traditionally been used in many parts of the world as alternative medicine. Many extracts and essential oils isolated from plants have disclosed biological activity, justifying the investigation of their potential antimicrobial activity. In this study, the in vitro antifungal activity of six Brazilian Cerrado medicinal plant species were evaluated against clinically relevant Candida species. METHODS: The crude extract plants were evaluated against American Type Culture Collection (ATCC) standard strains of Candida spp. using disk diffusion method and determining the minimum inhibitory concentration (MIC). The chemical study results were confirmed by HPLC method. RESULTS: All six plant species showed antifungal activity. Among the species studied, Eugenia dysenterica and Pouteria ramiflora showed significant inhibitory activity against C. tropicalis at lowest MIC value of 125 and 500 µg/disc, respectively. The Eugenia dysenterica also disclosed MIC value of 125 µg/disc against C. famata, 250 µg/disc against C. krusei and 500 µg/disc against C. guilliermondii and C. parapsilosis. Pouteria torta, Bauhinia rufa, Erythroxylum daphnites and Erythroxylum subrotundum showed activity against the yeast strains with MIC value of 1000 µg/disc. The chemical study of the most bioactive extracts of Eugenia dysenterica and Pouteria ramiflora revealed catechin derivatives and flavonoids as main components. CONCLUSIONS: All six evaluated plant species showed good antifungal potential against several Candida strains. However, E .dysenterica and P. ramiflora showed the higher inhibitory effect against the non-albicans Candida species. Our results may contribute to the continuing search of new natural occurring products with antifungal activity.

Radiation induced a supra-additive cytotoxic effect in head and neck carcinoma cell lines when combined with plant extracts from Brazilian Cerrado biome.

OBJECTIVES: Antineoplastic effects of molecules derived from plants have recently gained increasing attention as an additive to traditional therapies. The aim of this study was to evaluate the cytotoxic activity of plant extracts from the Brazilian Cerrado biome associated with radiotherapy in head and neck carcinoma cells (HNSCC). MATERIALS AND METHODS: Fifteen extracts derived from five Cerrado plants were tested in HNSCC cell lines (SCC-25, SCC-9, FaDu) and keratinocyte cells (HaCat). Cell cytotoxicity of extracts and association extract/radiation (2Gy/min) was assessed by MTT assay. Cisplatin (50 µg/mL) was used as a positive control. Extracts with the major cytotoxic activity were selected and their IC50 concentrations were defined. Apoptosis was assessed using flow cytometric analysis. RESULTS: Ten isolated extracts resulted in moderate cytotoxicity (>20 and ≤ 50 % of viable cells), while three extracts induced severe cytotoxic effects (≤ 20 % of viable cells). Plant extracts treatment improved radiotherapy cytotoxicity in all cell lines. Although plant extracts are not as potent as cisplatin plus radiation, in FaDu cells, seven extracts associated with irradiation showed cytotoxic activity similar or better than the association of cisplatin and radiation. Hexanic extract of Erythroxylum daphinites could induce apoptosis in oral cancer cells; however, necrosis was the prevalent kind of death in FaDu cells treated with hexanic extract of Erythroxylum suberosum. CONCLUSIONS: Pre-treatment of HNSCC cells with the extract derived from Cerrado plants followed by irradiation induced a supra-additive cytotoxic effect. CLINICAL RELEVANCE: This study highlights the potential biological relevance of the Cerrado biome when associated with traditional therapy for cancer.

Determination of rutin in Erythroxylum suberosum extract by liquid chromatography: applicability in standardization of herbs and stability studies / Determinación de rutina en el extracto Erythroxylum suberosum por cromatografía líquida: aplicación de la normalización de las hierbas y los estudios de estabilidad

In this study, a reverse phase-high performance liquid chromatography (RP-HPLC) technique for determination of rutin in Erythroxylum suberosum extract was developed and validated. A regression analysis was performed, with the observation of good linearity (r = 0.9997). The values obtained for precision and accuracy determination are in agreement with ICH guidelines. The detection and the quantitation limits of rutin were 0.19 ug/mL and 0.60 ug/mL, respectively. The results demonstrated that the developed method is a reliable HPLC technique for determination of rutin in E. suberosum extract. In addition, the applicability of this method in stability studies and standardization of herbs was investigated.

En este estudio, la técnica de cromatografía líquida de alta resolución en fase reversa para la determinación de la rutina en el extracto Erythroxylum suberosum fue desarrollada y validada. Se realizó un análisis de regresión, con la observación de una buena linealidad (r = 0,9997). Los valores obtenidos para la precisión y la determinación de la precisión están de acuerdo con las directrices ICH. La detección y cuantificación de los límites de la rutina fueron 0,19 ug / mL y 0,60 ug / mL, respectivamente. Los resultados demostraron que el método desarrollado es una técnica fiable de HPLC para la determinación de la rutina en el extracto de E. suberosum. Además, se investigó la aplicabilidad de este método en los estudios de estabilidad y la estandarización de hierbas.

Changes in the trade in native medicinal plants in Brazilian public markets.

Plants continue to be an important source of new bioactive substances. Brazil is one of the world's mega-diverse countries, with 20 % of the world's flora. However, the accelerated destruction of botanically rich ecosystems has contributed to a gradual loss of native medicinal species. In previous study, we have observed a fast and intensive change in trade of medicinal plants in an area of Amazon, where human occupation took place. In this study, we surveyed 15 public markets in different parts of Brazil in search of samples of 40 plants used in traditional medicine and present in first edition of Brazilian Official Pharmacopoeia (FBRAS), published in 1926. Samples of plants commercialized as the same vernacular name as in Pharmacopoeia were acquired and submitted to analysis for authentication. A total of 252 plant samples were purchased, but the laboratory analyses showed that only one-half of the samples (126, 50.2 %) were confirmed as the same plant species so named in FBRAS. The high number of unauthenticated samples demonstrates a loss of knowledge of the original native species. The proximity of the market from areas in which the plant occurs does not guarantee that trade of false samples occurs. The impact of the commerce of the substitute species on their conservation and in public health is worrying. Strategies are necessary to promote the better use and conservation of this rich heritage offered by Brazilian biodiversity.