Effects of Bryophyllum pinnatum Administration on Wistar Rat Pregnancy: Biochemical and Histological Aspects.

INTRODUCTION: Bryophyllum pinnatum is widely used in folk medicine. It has neuropharmacological, anti-inflammatory, immunomodulatory, antidiabetic, hepatoprotective, and nephroprotective effects, among others. It also acts on uterine contractility. It is prescribed by practitioners of anthroposophic medicine for preterm labor, insomnia, and emotional disorders, and has other potential uses in obstetrics. As all drugs currently used in preterm labor have side effects, new tocolytic agents remain an area of active research. OBJECTIVE: To evaluate the effect of B. pinnatum mother tincture (MT) on albino rats and their offspring throughout pregnancy from a biochemical and histological standpoint. METHODS: Longitudinal, prospective, randomized controlled bioassay. This is the second stage of a trial that investigated 60 animals distributed across six equal groups: controls C1 and C2, which received 1 and 25 times the vehicle dose (30% ethanol), B1 and B2 (1- and 25-fold doses of B. pinnatum MT), and B3 and B4 (which received 50- and 100-fold doses of B. pinnatum concentrate). At this stage, blood chemistry parameters (glucose, alanine aminotransferase [ALT], aspartate aminotransferase [AST], creatinine, and blood urea nitrogen) were measured in dams, as well as histological aspects of dam liver, kidney, placenta, and uterine tissue and fetal liver, kidney, heart, and brain. RESULTS: No differences were found between group B1 (therapeutic dose) and its control C1 in relation to glucose, AST, ALT, and creatinine. Group B2 exhibited lower glucose levels than groups C1, B3, and B4. There was no difference in AST across groups. Groups B3 and B4 exhibited higher ALT levels than groups C1 and B1. Groups B1-B4 exhibited higher urea nitrogen levels than group C1. Creatinine levels were higher in groups B2 and B3 than group C1. On morphological evaluation, fatty infiltration of the liver was observed in the alcoholic vehicle control groups (C1 and C2). CONCLUSIONS: Daily administration of B. pinnatum at therapeutic doses (group B1) to pregnant albino rats appears to be safe, with reduced glucose at dose B2, elevated ALT at doses B3 and B4, and increased urea at doses B1 to B4 and creatinine at B2 and B3, but never exceeding the normal reference range. It was not associated with histological changes in specimens of the maternal or fetal structures of interest.

Gastric Ulcer Healing Property of Bryophyllum pinnatum Leaf Extract in Chronic Model In Vivo and Gastroprotective Activity of Its Major Flavonoid.

Gastric ulcer is a common disease that develops complications such as hemorrhages and perforations when not properly treated. Extended use of drugs in the treatment of this pathology can provoke many adverse effects. Therefore, finding medicinal plants with gastroprotective and mucosal healing properties has gained increasing interest. Bryophyllum pinnatum (Crassulaceae), popularly known in Brazil as "saião" or "coirama," has been used to treat inflammatory disorders. It is rich in flavonoids, and quercetin 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside-Bp1 is its major compound. In this study, we aimed to investigate ulcer healing properties of B. pinnatum against an acetic acid-induced chronic ulcer model and the gastroprotective activity of Bp1 against gastric lesions induced by ethanol and indomethacin. Ultrafast liquid chromatography was used to quantify the main compounds (mg/g of the extract)-quercetin 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (33.12 ± 0.056), kaempferol 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (3.98 ± 0.049), and quercetin 3-O-α-L-rhamnopyranoside (4.26 ± 0.022) and showed good linearity, specificity, selectivity, precision, robustness, and accuracy. In vivo studies showed that treatment with the extract at 250 and 500 mg/kg stimulated the healing process in the gastric mucosa with significant ulceration index reduction, followed by improvement in the antioxidant defense system [increased glutathione (GSH) levels, decreased superoxide dismutase upregulation, and malondialdehyde (MDA) levels]. Moreover, the extract decreased interleukin-1ß and tumor necrosis factor-a levels and myeloperoxidase (MPO) activity, increased interleukin 10 levels, showed a cytoprotective effect in histological analyzes and also downregulated the expression of cyclooxygenase-2 and NF-κB (p65). The pretreatment with Bp1 at a dose of 5 mg/kg reduced gastric lesions in the ethanol and indomethacin models, increased GSH, and decreased MDA levels. In addition, the pretreatment decreased MPO activity, interleukin-1ß and tumor necrosis factor-α levels, while also showing a cytoprotective effect in histological analyzes. Our study suggests that treatment with B. pinnatum extract showed a higher inhibition percentage than pretreatment with the Bp1. This might in turn suggest that Bp1 has gastroprotective activity, but other compounds can act synergistically, potentiating its effect. We conclude that B. pinnatum leaf extract could be a new source of raw material rich in phenolic compounds to be applied in food or medicine.

Wound healing cream formulated with Kalanchoe pinnata major flavonoid is as effective as the aqueous leaf extract cream in a rat model of excisional wound.

Leaves of Kalanchoe pinnata are used worldwide for healing skin wounds. This study aimed to develop and compare two creams containing a leaf aqueous extract of K. pinnata (KP; 6%) and its major flavonoid [quercetin 3-O-α-L-arabinopyranosyl-(1→2)-α-L-rhamnopyranoside] (0.15%). Both creams were topically evaluated in a rat excision model for 15 days. On the 12th day, groups treated with KP leaf-extract and KP major flavonoid creams exhibited 95.3 ± 1.2% and 97.5 ± 0.8% of healing, respectively (positive control = 96.7 ± 0.8%; negative control = 76.1 ± 3.8%). Both resulted in better re-epithelialization and denser collagen fibres. Flavonol glycosides are the main phenolics in KP leaf-extract according to HPLC-ESI-MS/MS analysis. KP major flavonoid plays a fundamental role in the wound healing. The similar results found for both creams indicate that the use of KP crude extract should be more profitable than the isolated compound.

Bryophyllum pinnatum markers: CPC isolation, simultaneous quantification by a validated UPLC-DAD method and biological evaluations.

Bryophyllum pinnatum (Lam.) Oken (Crassulaceae) is widely used as leaf juice or extracts in traditional medicine all over tropical areas, especially in Brazil, to relieve inflammation-associated symptoms. Flavonol glycosides with unusual sugar moiety are among the major metabolites. Nevertheless, there are not enough quality control studies that can contribute to authentication of B. pinnatum and determination of their markers. As it is also used as medicinal plant in several countries, it is necessary to provide data related to safety, efficacy and quality. In this context, this work aims to isolate the major flavonoids from B. pinnatum hydroethanolic extract, to validate a method to quantify the content of chemical markers and to evaluate their xanthine oxidase inhibition and antioxidant activity. The extract was submitted to centrifugal partition chromatography (CPC). The solvents system CyHex-EtOAc-EtOH-H2O, 0.5:9:3:5.5, v/v/v/v was selected by shake-flask method. Four flavonoids (quercetin 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (1), kaempferol 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (2), quercetin 3-O-α-L-rhamnopyranoside (3) and kaempferol 3-O-α-L-rhamnopyranoside (4)) were isolated in a single and fast CPC run and their structures were confirmed by NMR analysis. An UPLC-DAD quantification method was established for the first time with validation of required parameters, according to RDC 166/2017. The calibration curves were linear with correlation coefficient ranging from 0.9996 to 0.9997 while the values of LOD (0.0077-1.984 ng.mL-1), LOQ (0.0263-6.012 ng.mL-1), recovery (≥ 80.7 %) and inter-day (%RSD ≤ 3.581) and intra-day precision (%RSD ≤ 2.628) were satisfactory. Quantitative analysis of these compounds showed that the proportion of 1, 2 and 3 were 2.43, 0.25 and 0.33 % (24.3 mg.g-1, 0.25 mg.g-1 and 0.33 mg.g-1 of extract), respectively. Moreover, in vitro xanthine oxidase (XO), DPPH and ABTS inhibition were evaluated for the extract and the major flavonoids. Compounds 2 (168 µM) and 3 (124 µM) moderately inhibited XO, while compounds 1 and 3 displayed average radical scavenging activity. In conclusion, our results suggest the flavonoid 1 as a specific marker which may be used for quality control of B. pinnatum hydroethanolic leaves extract.

Fatty acid compositions of ether extracts of Bryophyllumpinnatum Lam., Ficus exasperata Vahl., Gossypium herbaceum Linn. and Hilleria latifolia (Lam.) H. Walt / Composición de ácidos grasos de extractos etéreos de Bryophyllum pinnatum Lam., Ficus exasperata Vahl., Gossypium herbaceum Linn. y Hilleria latifolia (Lam.) H. Walt

SUMMARY Currently, there is a growing interest in identifying alternative sources of fatty acids due to the present increasing demand for oil-rich botanicals in industrial applications. The main objective of this work was to identify the fatty acid compositions of ether extracts of dried leaves of Bryophyllum pinnatum Lam., Ficus exasperata Vahl., Gossypium herbaceum Linn. and Hilleria latifolia (Lam.) H. Walt. The fatty acids were analyzed by gas chromatography with flame ionization detector. Among the evaluated ether extracts, the higher contents of saturated fatty acids were found in H. latifolia (27.96%) with the principal presence of stearic acid compared to the content of the saturated fatty acids in B. pinnatum (0.53%), F. exasperata (0.04%) and G. herbaceum (0.47%). Equally, the result showed that H. latifolia contained the highest percentage of unsaturated fatty acids with the predominant presence of oleic acid with the amount of 41.04%. Linoleic acid was also found to have the highest value in H. latifolia with the amount of 20.41%. Stearic acid, oleic acid, and linoleic acid were found in all the samples. The extract of H. latifolia contained a healthy mixture of different types of fatty acids thus suggesting it as a probable source of suitable fatty acids.

RESUMEN Actualmente, existe un creciente interés en identificar fuentes alternativas de ácidos grasos debido a la creciente demanda actual de productos botánicos ricos en aceite en aplicaciones industriales. El objetivo principal de este trabajo fue identificar las composiciones de ácidos grasos de extractos de éter de hojas secas de Bryophyllum pinnatum Lam., Ficus exasperata Vahl., Gossypium herbaceum Linn. y Hilleria latifolia (Lam.) H. Walt. Los ácidos grasos fueron analizados por cromatografía de gases con detector de ionización de llama. Entre los extractos de éter evaluados, los mayores contenidos de ácidos grasos saturados se encontraron en H. latifolia (27,96%) con presencia principal de ácido esteárico en comparación con el contenido de los ácidos grasos saturados en B. pinnatum (0,53%), F. exasperata (0,04%) y G. herbaceum (0,47%). Igualmente, el resultado mostró que la H. latifolia contenía el mayor porcentaje de ácidos grasos insaturados con presencia predominante de ácido oleico con una cantidad de 41,04%. También se encontró que el ácido linoleico tiene el valor más alto en H. latifolia con una cantidad de 20,41%. Se encontró ácido esteárico, ácido oleico y ácido linoleico en todas las muestras. El extracto de H. latifolia contenía una mezcla saludable de diferentes tipos de ácidos grasos, lo que sugiere que es una fuente probable de ácidos grasos adecuados.

Identification of a Selective PDE4B Inhibitor From Bryophyllum pinnatum by Target Fishing Study and In Vitro Evaluation of Quercetin 3-O-α-L-Arabinopyranosyl-(1→2)-O-α-L-Rhamnopyranoside.

Natural products are considered an important source of bioactive compounds especially in biodiversity-rich countries like Brazil. The identification of potential targets is crucial to the development of drugs from natural sources. In this context, in silico methodologies, such as inverse virtual screening (target fishing), are interesting tools as they are a rational and direct method that reduces costs and experimental time. Among the species of Brazilian biomes, Bryophyllum pinnatum (Lam.) Oken, native to Madagascar, is widely used by the population to treat inflammation conditions. It has a remarkable presence of flavonoids, including quercetin 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (1), considered one of its major compounds. However, until now there were no studies addressing its putative mechanism of action and explaining its pharmacological action. The enzyme PDE4B, known as an antiinflammatory protein, was indicated as a promising target by target fishing methods. This activity was confirmed by in vitro enzymatic inhibition, and an expressive selectivity of PDE4B over PDE4A was demonstrated. The interactions were investigated through molecular dynamics simulations. The results were pioneering, representing an advance in the investigation of the antiinflammatory action of B. pinnatum and confirm the potential of the flavonoid as a chemical extract marker. Also, the flavonoid was shown to be a promising lead for the design of other selective PDE4B blockers to treat inflammatory diseases.

Optimization of Aqueous Extraction from Kalanchoe pinnata Leaves to Obtain the Highest Content of an Anti-inflammatory Flavonoid using a Response Surface Model.

INTRODUCTION: The medicinal plant Kalanchoe pinnata is a phenolic-rich species used worldwide. The reports on its pharmacological uses have increased by 70% in the last 10 years. The leaves of this plant are the main source of an unusual quercetin-diglycosyl flavonoid (QAR, quercetin arabinopyranosyl rhamnopyranoside), which can be easily extracted using water. QAR possess a strong in vivo anti-inflammatory activity. OBJECTIVE: To optimize the aqueous extraction of QAR from K. pinnata leaves using a three-level full factorial design. MATERIAL AND METHODS: After a previous screening design, time (x1 ) and temperature (x2 ) were chosen as the two independent variables for optimization. Freeze-dried leaves were extracted with water (20% w/v), at 30°C, 40°C or 50°C for 5, 18 or 30 min. QAR content (determined by HPLC-DAD) and yield of extracts were analyzed. The optimized extracts were also evaluated for cytotoxicity. RESULTS: The optimal heating times for extract yield and QAR content were similar in two-dimensional (2D) surface responses (between 12.8 and 30 min), but their optimal extraction temperatures were ranged between 40°C and 50°C for QAR content and 30°C and 38°C for extract yield. A compromise region for both parameters was at the mean points that were 40°C for the extraction temperature and 18 min for the total time. CONCLUSION: The optimized process is faster and spends less energy than the previous one (water; 30 min at 55°C); therefore is greener and more attractive for industrial purposes. This is the first report of extraction optimization of this bioactive flavonoid. Copyright © 2018 John Wiley & Sons, Ltd.

Atividade antiulcerogênica do extrato aquoso da Bryophyllum pinnatum (Lam. ) Kurz / Antiulcerogenic activity of aqueous extract from Bryophyllum pinnatum (Lam. ) Kurz

Bryophyllum pinnatum (Lam.) Kurz pertence a família Crassulaceae e é conhecida vulgarmente como coirama, folha-da-fortuna, ou folha-do-ar, sendo usada popularmente como antifúngico, no tratamento da hipertensão, em úlceras e em inflamações. O presente estudo objetivou analisar a atividade antiulcerogênica do extrato aquoso das folhas de Bryophyllum pinnatum (Lam.) Kurz em modelo de úlcera gástrica induzida por indometacina em Rattus norvegicus, machos (150 a 250 g) com idade de 70 dias. Os animais receberam por via oral: água, extrato aquoso de Bryophyllum pinnatum (Lam.) Kurz (1 e 2 g Kg-1 ) ou ranitidina (60 mg Kg-1 ), fármaco com ação gastroprotetora conhecida que atua bloqueando os receptores de histamina H2. Após uma hora dos tratamentos, todos animais receberam indometacina via intraperitoneal. Os resultados demonstraram que o extrato aquoso da Bryophyllum pinnatum (Lam.) Kurz possui ação gastroprotetora; na dose de 1 g Kg-1 inibiu 45,49% o índice de ulceração induzido pela indometacina, e, na dose de 2 g Kg-1, inibiu 49,50%. Sugere-se o envolvimento de vários mecanismos na ação gastroprotetora dessa planta e não somente uma possível participação das prostaglandinas nesse efeito. Estudos futuros com diferentes modelos de indução de úlcera gástrica tornam-se necessários para melhor avaliar a atividade antiulcerogênica do extrato aquoso de Bryophyllum pinnatum (Lam.) Kurz.

Bryophyllum pinnatum (Lam.) Kurz belongs to the family Crassulaceae, is popularly known as "coirama", "folha-da-fortuna", or "folha-do-ar" and has been commonly used as antifungal, in the treatment of hypertension, ulcers and inflammation. The present study aimed to analyze the antiulcerogenic activity of aqueous extract from the leaves of Bryophyllum pinnatum (Lam.) Kurz using indomethacin-induced gastric ulcer models in Rattus norvegicus, males (150-250 g) aged 70 days old. The animals received by the oral route: water, aqueous extract from Bryophyllum pinnatum (Lam.) Kurz (1 and 2 g Kg-1) or ranitidine (60 mg Kg-1), a drug with known gastroprotective action, blocking histamine H2-receptors. After one hour of treatments, all animals received indomethacin by the intraperitoneal route. Results demonstrated that the aqueous extract from Bryophyllum pinnatum (Lam.) Kurz has gastroprotective action; at 1g kg-1 it inhibited 45.49% of the indomethacin-induced ulcer index, while at 2g Kg-1 it inhibited 49.50%. Several mechanisms of actions are suggested to be involved in the gastroprotective action of this plant, besides the possible participation of prostaglandins in this effect. Further studies using different gastric ulcer-induction models are needed to better evaluate the antiulcerogenic activity of aqueous extract from Bryophyllum pinnatum (Lam.) Kurz.

Avaliação da atividade antimicrobiana in vitro da planta Bryophyllum pinnatum Kurz ("Folha-da-fortuna") colhida em Várzea Grande, Mato Grosso/Brazil

As folhas de Bryophyllum pinnatum Kurz. (Folha-da-fortuna), Crassulaceae, têm sido usadas na medicina popular como emolientes para furúnculos, em queimaduras ou outros ferimentos, além de problemas respiratórios e gástricos. O trabalho teve por objetivo avaliar a atividade antimicrobiana do decocto de B. pinnatum Kurz. (Crassulaceae), sobre bactérias Gram positivas e Gram negativas. A avaliação da atividade antimicrobiana das folhas da planta em estudo foi realizada utilizando decocto, através da técnica de diluição em tubos, frente a três bactérias isoladas de cães com lesões de pele (Staphylococcus sp., Pseudomonas aeruginosa e Streptococcus sp.) e uma de fezes de suínos (Escherichia coli). O decocto apresentou, após 30 minutos de incubação, capacidade de inibir o crescimento bacteriano na concentração de 4mg/mL para Staphylococcus sp. Quando avaliado frente ao Streptococcus sp., não foi observada turvação do meio, embora tenha se observado um precipitado em todos os tubos. Na análise realizada com P. aeruginosa e E. coli, não foi observada inibição do crescimento em nenhuma diluição e tempo de ação do decocto. Os resultados demostraram uma ação antimicrobiana do decocto da folha de Bryophillum pinnatum Kurz. sobre bactérias Gram positivas.

Avaliação da atividade antimicrobiana in vitro da planta Bryophyllum pinnatum Kurz ("Folha-da-fortuna") colhida em Várzea Grande, Mato Grosso/Brazil

As folhas de Bryophyllum pinnatum Kurz. (Folha-da-fortuna), Crassulaceae, têm sido usadas na medicina popular como emolientes para furúnculos, em queimaduras ou outros ferimentos, além de problemas respiratórios e gástricos. O trabalho teve por objetivo avaliar a atividade antimicrobiana do decocto de B. pinnatum Kurz. (Crassulaceae), sobre bactérias Gram positivas e Gram negativas. A avaliação da atividade antimicrobiana das folhas da planta em estudo foi realizada utilizando decocto, através da técnica de diluição em tubos, frente a três bactérias isoladas de cães com lesões de pele (Staphylococcus sp., Pseudomonas aeruginosa e Streptococcus sp.) e uma de fezes de suínos (Escherichia coli). O decocto apresentou, após 30 minutos de incubação, capacidade de inibir o crescimento bacteriano na concentração de 4mg/mL para Staphylococcus sp. Quando avaliado frente ao Streptococcus sp., não foi observada turvação do meio, embora tenha se observado um precipitado em todos os tubos. Na análise realizada com P. aeruginosa e E. coli, não foi observada inibição do crescimento em nenhuma diluição e tempo de ação do decocto. Os resultados demostraram uma ação antimicrobiana do decocto da folha de Bryophillum pinnatum Kurz. sobre bactérias Gram positivas.

Avaliação da atividade antimicrobiana in vitro da planta Bryophyllum pinnatum Kurz ("Folha-da-fortuna") colhida em Várzea Grande, Mato Grosso/Brazil

As folhas de Bryophyllum pinnatum Kurz. (Folha-da-fortuna), Crassulaceae, têm sido usadas na medicina popular como emolientes para furúnculos, em queimaduras ou outros ferimentos, além de problemas respiratórios e gástricos. O trabalho teve por objetivo avaliar a atividade antimicrobiana do decocto de B. pinnatum Kurz. (Crassulaceae), sobre bactérias Gram positivas e Gram negativas. A avaliação da atividade antimicrobiana das folhas da planta em estudo foi realizada utilizando decocto, através da técnica de diluição em tubos, frente a três bactérias isoladas de cães com lesões de pele (Staphylococcus sp., Pseudomonas aeruginosa e Streptococcus sp.) e uma de fezes de suínos (Escherichia coli). O decocto apresentou, após 30 minutos de incubação, capacidade de inibir o crescimento bacteriano na concentração de 4mg/mL para Staphylococcus sp. Quando avaliado frente ao Streptococcus sp., não foi observada turvação do meio, embora tenha se observado um precipitado em todos os tubos. Na análise realizada com P. aeruginosa e E. coli, não foi observada inibição do crescimento em nenhuma diluição e tempo de ação do decocto. Os resultados demostraram uma ação antimicrobiana do decocto da folha de Bryophillum pinnatum Kurz. sobre bactérias Gram positivas.