Chemical constituents from Kalanchoe hybrida and their cytotoxicity / 中草药·英文版

Objective: Kalanchoe hybrida (Crassulaceae) is naturalized throughout all the island of Taiwan in China. The preliminary bioassay-guided fractionation of the crude extract of K. hybrida exhibited that the chloroform and n-butanol fractions possessed potent cytotoxicity against MCF-7, NCI-H460, and SF-268 tumor cell lines at 50 µg/mL concentration. Therefore, K. hybrida was selected as a target and the chemical constituents from the chloroform and n-butanol fractions of the crude extracts of K. hybrida were identified. The potential constituents were examined for their cytotoxicity against the tumor cell lines. Methods: A combination of conventional chromatographic techniques was performed on the crude extract of K. hybrida. The chemical structures of the purified constituents were identified on the basis of spectroscopic and spectrometric analysis. Results: The purification results had led to the characterization of totally 37 compounds. The isolated compounds 1, 2, and 4–12 were examined for their cytotoxicity in vitro, and bufadienolides 4–8 and flavonol glycoside 11 displayed significant cytotoxicity towards all the tested tumor cell lines among these tested compounds. Conclusion: The results indicated that these principles should be responsible for the bioactivity of corresponding partial fractions. The potential constituents could be further investigated to explore the new natural lead drugs.

Mecanismos de ação relacionados à atividade antiúlcera de Kalanchoe pinnata (Lam.) Pers. (Crassulaceae) / Mechanisms of action underlying antiulcer activity of Kalanchoe pinnata (Lam.) Pers. (Crassulaceae).

Kalanchoe pinnata (Lam.) Pers. (Crassulaceae) é uma espécie muito empregada na medicina tradicional no Brasil e em outras partes do mundo, especialmente Índia, países da África e China. É indicada popularmente para diversos fins incluindo o tratamento de úlceras gástricas. A análise fitoquímica revelou a presença de vários constituintes, em especial os flavonoides. O tratamento de úlcera gástrica convencional apresenta diversos efeitos colaterais e, na maioria das vezes, não evita a recidiva da lesão. Dessa maneira, é interessante encontrar uma terapêutica mais segura e efetiva. Com o objetivo de avaliar a segurança, foi realizado ensaio de citotoxicidade do extrato bruto, in vitro, com valor de IC50 igual a 0,926 mg/mL, sendo possível predizer um valor de LD50 (1341,46 mg/kg). Já em relação ao ensaio de citotoxicidade, in vitro, da fração acetato de etila não foi encontrado um valor de IC50. Resultados de fototoxicidade, in vitro, mostraram que o extrato bruto e fração acetato de etila de K. pinnata não possuem potencial fototóxico. A contagem microbiana na droga vegetal para bactérias aeróbias/mesófilas foi de 6,9 x 104 UFC/g e a contagem de bolores e leveduras foi de 2,4 x 103 UFC/g, ambos valores dentro do limite estabelecido pela OMS. Análise de endotoxinas também foi realizada para o extrato bruto (<4,0.105 UE/kg) e fração acetato de etila (<2,7.105 EU/kg) de K. pinnata. Referente à fitoquímica, diversos flavonoides foram identificados no extrato bruto e fração acetato de etila de K. pinnata. Paralelamente ao estudo fitoquímico foi verificado que a atividade gastroprotetora do extrato bruto envolve a ação das prostaglandinas e grupamentos sulfidrila. Já o mecanismo de gastroproteção da fração acetato de etila é dependente de prostaglandinas e óxido nítrico. A atividade cicatrizante do extrato bruto de K. pinnata também foi avaliada. De acordo com os resultados macroscópicos, as doses de 200mg/kg e 400 mg/kg reduziram a área de lesão, com uma taxa de 33% e 39%, respectivamente, após 7 dias de tratamento (p<0,05). Análise histológica dos grupos tratados com o extrato bruto (200 e 400 mg/kg) indicou melhor recuperação da lesão, verificada pela regeneração da mucosa gástrica e pelo restabelecimento da arquitetura glandular. As enzimas antioxidantes (catalase, superóxido dismutase e glutationa peroxidase) e a expressão de VEGF foram avaliadas no mecanismo de cicatrização de úlceras gástricas. Os resultados mostraram que a atividade antiulcerogênica foi mediada pela ação antioxidante da enzima SOD. Não foi evidenciado in vivo o aumento da expressão de VEGF e nem o sequestro do radical peroxil nos animais tratados com o extrato bruto. Os resultados dos ensaios in vitro (ORAC) mostraram uma maior capacidade de sequestro de radicais peroxil da fração acetato de etila (1192,35 ± 112,61 µmol equivalente de Trolox/g de amostra seca) quando comparado com o extrato bruto (431,32 ± 7,17 µmol equivalente de Trolox/g de amostra seca). A atividade anti Helicobacter pylori também foi avaliada, no entanto, o extrato bruto não apresentou atividade anti H.pylori. Ademais, o extrato bruto demonstrou um potencial anti-inflamatório, pois foi observada uma redução nos níveis de TNF-α e L-selectina, após o tratamento em neutrófilos estimulados com LPS. Analisando os resultados sugere-se que K. pinnata possui um potencial terapêutico no combate de úlceras gástricas e possivelmente, anti-inflamatório, sendo que os flavonoides podem estar relacionados com o efeito biológico observado.

Kalanchoe pinnata (Lam.) Pers. (Crassulaceae) is a commonly used species in traditional medicine in Brazil and in other parts of the world, especially India, Africa and China, for the treatment of various diseases, including gastric ulcers. Phytochemical analysis revealed the presence of several constituents in this plant, especially flavonoids. The available pharmaceutical products to treat peptic ulcer have several side effects and, in most cases, do not prevent recurrence of the gastric lesions. Therefore, it is important to find a safer and more effective therapy. In order to evaluate safety, the in vitro cytotoxicity assay of crude extract from K. pinnata was performed. The IC50 value was 0,926 mg/mL corresponding to LD50 value (1341, 46 mg/kg). It was not determined IC50 value in vitro cytotoxicity assay for ethyl acetate fraction from K. pinnata. Neither the crude extract nor ethyl acetate fraction from K. pinnata showed phototoxicity. Microbial counting was performed on the K. pinnata-based drug in order to investigate microbiological contamination. The microbial count for aerobic / mesophilic bacteria was 6.9 x 104 CFU/g, and yeast count was 2.4 x 103 CFU/g, both values in agreement with the limits established by WHO. Endotoxin analysis was also performed for the crude extract (<4,0.105 UE/kg) and for ethyl acetate fraction (<2,7.105 UE/kg) from K. pinnata. In the phytochemical analysis several flavonoids were identified in the crude extract and ethyl acetate fraction of K. pinnata. In parallel to the phytochemical study, it was verified that the gastroprotective activity of the crude extract of K. pinnata involved prostaglandins and sulfhydril compounds. On the other hand, the mechanism of gastroprotection of the ethyl acetate fraction of K. pinnata is dependent on prostaglandins and nitric oxide. The healing activity of the crude extract of K. pinnata was also evaluated. According to the macroscopic results the dose of 200 mg/kg and 400mg/kg reduced the injury area, with a rate of 33% and 39%, respectively, after 7 days of treatment (p <0.05). Histological analysis showed regeneration of the gastric mucosa and re-establishment of the glandular architecture in groups treated with the crude extract (200 and 400 mg/kg). Antioxidant enzymes (catalase, superoxide dismutase and glutathione peroxidase) and VEGF expression were evaluated in the mechanism of gastric ulcer healing. The results showed that the antiulcerogenic activity was mediated by SOD. It was not demonstrated an increase in VEGF expression and nor in the in vivo sequestration of the peroxyl radical in the animals treated with crude extract. The results of in vitro assay (ORAC) showed a greater sequestering of peroxyl radical to the ethyl acetate fraction (1192,35 ± 112,61 µmol equivalent of Trolox/g of ethyl acetate fraction) when compared to the crude extract (431,32 ± 7,17 µmol equivalent of Trolox/g of crude extract) of K. pinnata. The anti Helicobacter pylori activity was also evaluated; however, the crude extract did not show anti H. pylori activity. However, the crude extract of K. pinnata demonstrated an anti-inflammatory potential, because TNF-α and L-selectin levels were reduced after treatment in LPS-stimulated neutrophils. The analysis of the results suggests that K. pinnata has a therapeutic potential against gastric ulcers and possible anti-inflammatory properties, and the flavonoids may be linked to the biological effect.

Flavonoid Compounds from the Leaves of Kalanchoe prolifera and Their Cytotoxic Activity against P-388 Murine Leukimia Cells

Seven flavonoid compounds, kaempferol (1), quercetin (2), quercetin-3-O-β-D-glucopyranoside (3), kaempferol-3-O-β-D-glucopyranoside (4), kaempferol-3-O-α-L-rhamnoside (5), quercetin-3-O-sophoroside (6) and quercetin-3-O-rutinoside (7), were isolated from the methanolic extract of leaves of Kalanchoe prolifera. Compounds 1-7 were isolated for first time from this plant. These compounds were evaluated their cytotoxic activity against P-388 murine leukimia cells in vitro. Among those compounds kaempferol (1) and quercetin (2) showed strongest cytotoxic activity with IC₅₀ values of 4.45 ± 0.05 and 6.28 ± 0.02 µg/mL, respectively.

Influence of Benzyladenine and Thidiazuron on Shoot Regeneration from Leaf and Shoot tip Explants of Sedum sarmentosum Bunge

Effects of N6-benzyladenine (BA) or thidiazuron (TDZ) on adventitious shoot regeneration and axillary shoot multiplication of Sedum sarmentosum was investigated. Leaf and shoot tip explants obtained from in vitro-grown shoots of S. sarmentosum were cultured on Murashige and Skoog (MS) medium supplemented with 0, 2.0, 4.0 or 8.0 µM BA or TDZ. Of the two cytokinins studied, BA was found to be more responsive as compared to TDZ with respect to number of shoots produced per explant. High frequency of shoot regeneration (92.2%) with a mean of 12.3 shoots was produced when the leaf explants were cultured on MS medium supplemented with 8.0 µM BA. The highest number of shoots (25.4) was obtained when shoot tip explants were cultured on MS medium devoid of cytokinins after 35 days of culture. For root induction, regenerated shoots were cultured on MS medium supplemented with 0, 2.0, 4.0 or 8.0 µM indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthaleneacetic acid (NAA). The highest number of (17.6) roots per shoot was obtained on MS medium supplemented with 2.0 µM IBA after 30 days of culture. Regenerated plantlets were successfully acclimatized in the greenhouse with 100% survival rate.

Genus Kalanchoe (Crassulaceae): A Review of Its Ethnomedicinal, Botanical, Chemical and Pharmacological Properties.

Genus Kalanchoe comprises hundred species. Different extracts of these Kalanchoe species have been widely used in traditional medicine. Recently it has been reported that Kalanchoe extracts possess various biological activities viz. antiviral, sedative, antiulcer, immunomodulatory, antileishmanial, CNS depressant, anti-inflammatory, thyroid peroxidase inhibitor, cytotoxic, hepatoprotective, antioxidant, analgesic, anticonvulsant, antimicrobial, inhibition of B cell development, cardiovascular, antihyperglycemic, acetylcholinesterase inhibition, insecticidal and larvicidal activities. Earlier studies on different Kalanchoe species have reported the isolation of polysaccharides, flavonoids, sterols, ascorbic acid, trace elements, organic acids, hydrocarbons, triterpenoids, phenolic components and bufadenolides. This review presents the botany, chemistry, traditional uses and pharmacological data of genus Kalanchoe.

Chemical constituents of roots Rhodiola crenulata / 中国药学杂志

OBJECTIVE: To study the chemical constituents of the roots of Rhodiola crenulata. METHODS: The roots of R. crenulata were extracted with 80% ethanol, and the extract was partitioned with ethyl acetate. The chemical components were isolated by various chromatographic methods. Their structures were elucidated by spectral analysis. RESULTS: Eleven compounds were isolated and identified as isolariciresinol-9, 9′-acetonide (1), p-coumaric acid (2), p-coumaric acid-4-O-β-D-glucopyranoside (3), tachio-side (4), dihydroconiferin (5), 2-phenylethyl β-glucopyranoside (6), 2-phenylethyl β-vicianoside (7), 2-phenylethyl-6-0-α -L-arabi-nofuranosyl-β-D-glucopyranoside(8), 2-phenylethyl β-primeveroside (9), benzyl β-D-glucopyranoside (10), and rhodiocyanoside D (11). CONCLUSION: All of the compounds were isolated from Rhodiola crenulata for the first time, and compounds 1, 3, 4, 5, 7, 9 were obtained from the genus of Rhodiola for the first time.

Chemical constituents in herbs of Bryophyllum pinnatum / 中草药

Objective: To isolate and identify the chemical constituents from the herbs of Bryophyllum pinnatum. Methods: The compounds were isolated by chromatography on silica gel and Sephadex LH-20 columns and preparative TLC. The structures of the isolated compounds were elucidated based on spectroscopic data and chemical properties. The anti-allergic activities of the compounds were evaluated by RBL-2H3 cells. Results: Six compounds were isolated from the herbs of B. pinnatum. Their structures were identified as bryophylluside (1), kaempfrol (2), rutin (3), quercetin 3-O-α-L-arabinopyranosyl (1→2) α-L-rhamnopyranoside (4), quercitrin (5), and β-sitosterol (6). Conclusion: Compound 1 is a new lignan, and lignans are first isolated from B. pinnatum. Kaempfrol is one of the main anti-allergic active compounds in B. pinnatum.

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.

Kalanchoe thrysiflora Harv. and Kalanchoe marmorata Baker; DNA Profiling, biological guided fractionation of different extracts; isolation and identification of cytotoxic compounds.

DNA profiling of two closely related ornamental plants belonging to family crassulaceae viz. Kalanchoe thrysiflora Harv. and Kalanchoe marmorata Baker were performed to establish genetic polymorphism. Biological guided fractionation of the two plant extracts to assess their cytotoxicity, had led to the isolation of one steroidal and one triterpenoidal compound from the most active dichloromethane fraction of Kalanchoe thrysiflora. The cytotoxicity of the isolated compounds were evaluated against normal (HFB4) and cancer (MCF7) cells. Compound 1 (3-oxo-olean-12-ene) and compound 2 (β-sitosterol) showed similar cytotoxic activity on MCF7 at IC50 17.4 and 17.6 μg/ml respectively while on HFB4, the compounds revealed cytotoxic activity at IC50 21.9 and 21.6 respectively.

Phytochemical profile of the stems of Aeonium lindleyi

Aeonium species are plants rich in phenols and terpenoids and have been widely used in the Canary folk medicine. Following our project concerning bioactive compounds from Macaronesian region, a phytochemical study was carried out to investigate the constituents of the stems of Aeonium lindleyi Webb & Berthel.,Crassulaceae. Air dried and chopped stems of this plant were extracted with ethanol in a Soxhlet apparatus and the total extract was chromatographed on silica gel to afford sixteen known compounds including, one sterol, four lupane triterpenes and eleven phenolic compounds. Their structures were elucidated by means of ¹H- and 13C-NMR spectroscopic studies and comparison of their spectral data with values in the literature. Both group of isolated metabolites are known by their diverse biological activities and they have been described as antioxidant, cytotoxic, anti-inflammatory. Our results contribute to the knowledge of A. lindleyi as a potential source of bioactive compounds.

Atividade inibitória das folhas e caule de Kalanchoe brasiliensis Cambess frente a microrganismos com diferentes perfis de resistência a antibióticos / Inhibitory activity of aerial parts of Kalanchoe brasiliensis Cambess against microorganisms with variation profile antibiotic-resistant

A avaliação antimicrobiana das partes aéreas de Kalanchoe brasiliensis Cambess, Crassulaceae, a qual é comumente utilizada para o tratamento de inflamações da mucosa oral, bronquites e congestão nasal, é relatada. Esta atividade foi avaliada em discos de Petri usando o método de difusão para a determinação da Concentração Inibitória Mínima (MIC) e cinética bactericida. Foram usadas amostras bacterianas gram-positivas, gram-negativas e cepas de fungos leveduriformes do gênero Cândida. Apenas o óleo essencial demonstrou ser efetivo, apresentando atividade frente amostras gram-positivas de Staphylococcus aureus (MRSA) meticilina resistente. A ação observada foi considerada bacteriostática por reduzir um log10 UFC/mL a partir da sexta hora de exposição da amostra ao óleo essencial nas concentrações de 4 por cento e 8 por cento. Compostos fenólicos estão presentes em óleo essencial, sugerindo que o efeito foi devido à presença dos mesmos. Por este motivo à planta Kalanchoe brasiliensis Cambess pode representar uma alternativa terapêutica para infecções provocadas por Staphylococcus aureus.

This study reports the antimicrobial evaluation of the aerial parts of Kalanchoe brasiliensis Cambess, Crassulaceae, commonly used for the treatment of the oral mucosa inflammation, bronchitis and nasal congestion. The antimicrobial activity was assayed in petri dishes using the diffusion method for determination of the minimal inhibitory concentration (MIC) and the kill curve kinetic methods. It were used gram-positive and gram-negative strain, leveduriforms fungi strain classified in genus Cândida. Only the essential oil showed activity against methicilin resistant Staphylococcus aureus (MRSA). This action was considered bacteriostatic with the reduction to one log10 CFU/ml after six hour of exhibition at the concentration of 4 percent and 8 percent. There are studies accounts that polyphenols are present in the essential oil and are active against bacteria. K. brasiliensis is rich in polyphenols suggering that the antimicrobial effect showed is due to this. For this reason, the plant Kalanchoe brasiliensis, can represent a therapeutic alternative against infections caused for Staphylococcus aureus.