Isolation and identification of chemical constituents of C14 pterosins from Pteris cretica / 中草药

Objective: To study the chemical constituents of pterosins from Pteris cretica. Methods: According to the characteristics of UV absorption and MSn fragmentation of pterosins, the pterosins were isolated and purified by silica gel, Sephadex LH-20 column and C18 semi-preparative column chromatography, and their structures were identified by physicochemical properties and spectral data. Results: A total of 19 pterosins with C14 were isolated and their structures were identified as (2S,3S)-pterosin T (1), (2S,3S)-pterosin C 3-O-β-D-glucoside (2), (2R,3S)-pterosin C 3-O-β-D-glucoside (3), (2S)-pterosin B 14-O-β-glucoside (4), 6-(1,2-dihydroxyethyl)- 2,5,7-trimethyl-1-indene-1-one (5), (2R,3S)-pterosin C (6), (2S,3S)-pterosin C (7), (2S)-pterosin P (8), (2R,3S)-pterosin S (9), (2S,3S)-pterosin S (10), (2R,3S)-pterosin Q (11), (2S,3S)-pterosin Q (12), (2S,3S)-pterosin C 14-O-β-D-glucoside (13), (2R,3S)-pterosin C 14-O-β-D-glucoside (14), (2S,3S)-pterosin S 14-O-β-D-glucoside (15), dehydropterosin B (16), (2R,3S)-pterosin Q 3-O-β-D-glucoside (17), (2S,3S)-pterosin Q 3-O-β-D-glucoside (18), and (2S,3S)-pterosin T 14-O-β-D-glucoside (19). Conclusion: Compounds 2-5, 10, 11, 14-19 are isolated from Pteris cretica for the first time.

Study on Chemical Constituents of Petroleum Ether Part from Whole Herbs of Pteris vittata / 中国药房

OBJECTIVE： To study the chemical constituents from the petroleum ether part from whole herbs of Pteris vittata. METHODS： The petroleum ether part from whole herbs of P. vittata was separated and isolated by silica gel column， gel column， recrystallization and TLC. The structures of the compounds were identified according to physicochemical properties and spectrum data （1H-NMR and 13C-NMR）. RESULTS： A total of 11 compounds were isolated and identified from the petroleum ether part from whole herbs of P. vittata， as （2R）-acetyl pterosin B （Ⅰ）， palmitic acid （Ⅱ）， hop-22（29）-ene （Ⅲ）， epifriedelanol （Ⅳ）， lupenone （Ⅴ）， olean-18-en-3-one （Ⅵ）， stigmasterol （Ⅶ）， β-sitosterol （Ⅷ）， 22-hydroxyhopane （Ⅸ）， ergosterol （Ⅹ）， β-sitosterol acetate （Ⅺ）. CONCLUSIONS： Compounds Ⅰ-Ⅶ and Ⅸ-Ⅺ are isolated from this plant for the first time， and can provide theoretic reference for further studying bioactive pharmacodynamic substances in P. vittata and enriching chemical component data.

Study on chemical constituents from Pteris semipinnata / 中草药

Objective To study the chemical constituents from the whole plants of Pteris semipinnata. Methods The chemical constituents were isolated by silica gel, sephadex LH-20, ODS, macroporous resin, MCI Gel resin and semi-preparation HPLC. Their structures were determined on the basis of NMR and MS analysis. Results Twenty compounds were isolated and identified as apigenin (1), apigenin-7-O-β-D-glucopyranoside (2), apigenin-7-O-β-D-gentiobioside (3), apigenin-7-O-β-D-glucopyranosyl-4′-O-α- L-rhamnopyranoside (4), isoviolanthin (5), luteolin (6), luteoloside (7), luteolin-7-O-β-D-gentiobioside (8), quercetin-3-O-β-D- glucopyranoside (9), kaempferol-3-O-β-D-glucopyranoside (10), rutin (11), epigallocatechin (12), pinoresinol-4-O-β-D- glucopyranoside (13), phillyrin (14), bergeninum (15), protocatechuic acid (16), trans-caffeic acid (17), gallic acid (18), β-sitosterol (19), and β-daucosterol (20). Conclusion Compounds 3, 5, 12, 13, and 15—18 are isolated from the plants of Pteris, and 2—5, 8—13, and 15—20 are isolated from this plant for the first time.

Chemical constituents from Pteris dispar and their anti-tumor activity in vitro / 中国中药杂志

The dried whole plant of Pteris dispar were milled and extracted with 95% EtOH. The resulting dried extract was isolated by kinds of chromatographic column, including polyamide, Sephadex LH-20, preparative HPLC. As a result, ten diterpenes were isolated from the plant. By analyzing of ESI-MS and NMR spectroscopic data, the structures were established as geopyxin B(1), geopyxin E(2), ent-11α-hydroxy-18-acetoxykaur-16-ene(3), ent-14β-hydroxy-18-acetoxykaur-16-ene(4), neolaxiflorin L(5), ent-3β,19-dihydroxy-kaur-16-ene(6), ent-3β-hydroxy-kaur-16-ene(7), 7β,17-dihydroxy-16α-ent-kauran-19-oic acid 19-O-β-D-glucopyranoside ester(8), crotonkinin C(9)and crotonkinin C(10). Compounds 1-10 were obtained from P. dispar for the first time. Compounds 1 and 2 showed moderate activities against Bel-7402 with IC₅₀ values of 7.50 and 10.60 μmol•L⁻¹, and against HepG2 with IC₅₀ values of 6.68,11.80 μmol•L⁻¹, respectively.

Development and validation for the determination of total flavonoids and luteolin from Pteris multifida Poir / 国际中医中药杂志

Objective To determine the content of total flavonoids and luteolin from Pteris multifida Poir. Methods The content of total flavonoids was determined by gradient elution of macroporous resin D101 and ultraviolet spectrophotometry. The content of luteolin was determined by HPLC. The analysis was performed on a RP-C18 column(4.6 mm×250 mm, 5 μm) with aceconitrile-0.2% phosphoric acid (35:65) as the mobile phase at a flow rate of 1.0 ml/min, and 30 ℃ temperature. Results The detection of wave length was set at 349 nm. The content of luteolin was 0.015%, 0.019%, 0.016%, and the content of total flavonoids was 0.015%, 0.019%, 0.016%, respectively. Conclusions The method is suitable for the determination of flavonoids componets from Pteris multifida Poir.

Chemical constituents from Pteris multifida and cytotoxic activity / 中国中药杂志

The materials were extracted by 95% ethanol, and the extracting solution was isolated by kinds of chromatographic columns including polyamide, MCI, preparative MPLC, and preparative HPLC. Eight diterpenes and two sesquiterpenes were isolated from the plant. On analysis of ESI-MS and NMR spectroscopic data, the structures were established as ent-3β-hydroxy-kaur-16-en-19-al (1), 4-epi-kaurenic acid (2), mitrekaurenone (3), 7β,16α,17-trihydroxy-ent-kauran-19-oic acid (4), crotonkinin E (5), crotonkinin F (6), pterisolic acid A (7), pterisolic acid C (8), (2R)-pterosin P (9), and dehydropterosin B (10). Compounds 1-6 were obtained from Pteris for the first time, and compounds 7-10 were obtained from P. ensiformis for the first time. Compounds 5-8 showed moderate activity against HCT-116, HepG2 and BGC-823 cell lines, separately.

Chemical constituents from Pteris deltodon / 中草药

Objective: To study the chemical constituents in the whole plant of Pteris deltodon. Methods: Nine compounds were isolated from 95% ethanol extract in the whole plant of P. deltodon and purified by silica gel chromatography, Sephadex LH-20, and pre-HPLC, and their structures were identified on the basis of spectroscopic data and literatures. Results: Nine compounds were isolated and elucidated as β-sitosteol (1), (22E)-5α,8α-epidioxyergosta-6,22-dien-3β-ol (2), ent-kaur-16-ene-2β,15α-diol (3), cycloart-23-en-3β,25-diol (4), cycloart-25-en-3β,24-diol (5), emodin (6), 1,7-dihydroxyxanthone (7), ursolic acid (8), and ergosterol (9). Conclusion: All the compounds are for the first time isolated from P. deltodon; Compound 4 and 5 are cycloartanes first isolated from genus Pteris L.

Actividad antimicrobiana in vitro de Pteris vittata L / In vitro antimicrobial activity of Pteris vittata L

Introducción: las hojas de Pteris vittata L (helecho) son utilizadas por la población para el tratamiento de la candidiasis y en enfermedades producidas por bacterias en la piel. Objetivo: identificar preliminarmente las familias de metabolitos secundarios presentes en las hojas de la planta y evaluar su posible actividad antimicrobiana. Métodos: se recolectaron las hojas de Pteris vittata L. El material vegetal fue lavado, desinfectado, secado y seguidamente se procedió a su pulverización. Este polvo se utilizó en la elaboración de los diferentes extractos y tintura. La tintura obtenida se concentró y se fraccionó sucesivamente con n-hexano, cloroformo y acetato de etilo. A estos extractos se les realizó el tamizaje fitoquímico, ensayos microbiológicos y cromatografía de capa fina. Resultados: las pruebas in vitro efectuadas a los extractos obtenidos a partir de la tintura 20 por ciento, demostraron que éstos presentan actividad antimicrobiana frente a Escherichia coli, Staphylococcus aureus, destacándose los resultados obtenidos frente a Candida sp para los extractos de acetato de etilo y clorofórmico. En estas fracciones están presentes en mayor proporción alcaloides y quinonas, que podrían ser los responsables de esta actividad, lo cual se corrobora con la identificación de estos metabolitos secundarios mediante la cromatografía de capa fina y el tamizaje fitoquímico realizado. Conclusiones: el estudio combinado mediante la cromatografía de capa fina y el tamizaje fitoquímico de los extractos hexánico, acetato de etilo y clorofórmico permite inferir que la actividad antimicrobiana puede deberse a la presencia de quinonas y alcaloides(AU)

Introduction: Pteris vittata L. leaves (fern) are used by people on the candidiasis treatment and some skin illnesses caused by bacteria. Objective: to identify preliminarily the secondary metabolites present in the leaves of the plant and to evaluate their possible antimicrobial activity. Methods: Pteris vittata L. leaves were collected. The plant material was washed, disinfected, dried and pulverized. The powder obtained was used to make the various extracts and the tincture. The latter was concentrated and successively fractionated with n-hexane, chloroform, and ethyl acetate. The extracts underwent phytochemical screening, microbiological assays and thin-layer chromatography. Results: in vitro tests performed in the obtained extracts from the 20 percent tincture proved that they have antimicrobial activity against Escherichia coli and Staphylococcus aureus, emphasizing the accomplished results against Candida of the ethyl and chloroform acetate extracts. Alkaloids and quinones, which are found in large proportion in the extracts, would be responsible of the above- mentioned antibacterial activity. This was corroborated by the identification of these secondary metabolites through thin-layer chromatography and phytochemical screening. Conclusions: the combined study through thin-layer chromatography and phytochemical screening of the ethyl and chloroform acetate extracts showed that the antimicrobial activity could be possible due to the alkaloids and quinones presence(AU)

In vitro Antioxidant and Anticancer Properties of Active Compounds from Methanolic Extract of Pteris multifida Poir. Leaves.

Aims: To isolate pure compounds from the methanolic fraction obtained from successive fractionation of defatted ethanolic extract and evaluate in vitro antioxidant and anticancer activity of the crude ethanolic extract, methanolic fraction and pure compounds isolated from methanolic fraction from leaves of Pteris multifida Poir. Study Design: Isolation and identification of the compounds, evaluation of antioxidant and anticancer activity on cervical cancer cell line (HeLa), lung carcinoma cell line (NCIH460) and breast carcinoma cell line (MCF-7). Place and Duration of Study: Vietnam Academy of Science and Technology of Ho Chi Minh City and School of Biotechnology, International University, Vietnam National University, Ho Chi Minh City, between December 2012 and September 2013. Methodology: The crude ethanolic leaf extract and methanolic fraction obtained from successive fractionation of defatted ethanolic extract from Pteris multifida leaves were prepared. The isolated compounds from methanolic fraction were identified using different spectroscopic techniques. Antioxidant activity of the samples was evaluated by using the stable free radical 2, 2- diphenyl picrylhydrazyl (DPPH). Sulforhodamine B (SRB) assay was exploited for determination of anticancer activity against three selected human cancer cell lines: HeLa, NCI-H460 and MCF-7. Results: Two main compounds were isolated from methanolic fraction obtained from successive fractionation of defatted ethanolic extract: rutin (1) and apigenin-7-O-β-Dglucopyranoside (2). The crude ethanolic leaf extract showed weak antioxidant activity (IC50 = 89.84 μg/mL) whereas the methanolic fraction expressed quite strong antioxidant activity (IC50 = 21.9 μg/mL). Rutin (1) showed a good ingredient of antioxidant activity with IC50 value of 37.70 ± 0.03 μg/mL. Crude ethanolic leaf extract had cytotoxic activity against HeLa and NCI-H460 cell lines while the methanolic fraction had cytotoxic activity against HeLa, NCI-H460 and MCF-7 cell lines. Apigenin-7-O-β-D-glucopyranoside (2) had strong anticancer activity against MCF-7 cell line with IC50 = 22.62 ± 0.59 μg/mL. Conclusion: The crude ethanolic leaf extract and its methanolic fraction of P. multifida showed the potential activity in antioxidant and anticancer activity. Rutin had a potent antioxidant activity while apigenin-7-O-β-D-glucopyranoside had a strong anticancer activity against the human breast adenocarcinoma cell line MCF-7.

In-vitro shoot regeneration via caulogenesis in fern, Pteris vittata L.

An attempt was made to standardize the protocol for the shoot regeneration via caulogenesis in Pteris vittata L. employing leaf primordium explants. Calli were induced on Murashige and Skoog’s (MS) and Parkers and Thompson’s (P and T) media supplemented with different combinations of 2, 4-dichlorophenoxyaceticacid (2, 4-D), 6-benzylaminopurine (BAP), a-naphthalene acetic acid (NAA) and Indole - 3-acetic acid (IAA). A combination of full strength MS medium with 2, 4-D (2.26 µM) and BAP (2.22 µM) was found to be ideal for profuse callusing (80%) against other combinations. More shoot differentiation (2.8±0.06) was achieved from the calli on one-fourth strength of P and T media fortified with BAP (4.44 µM) and NAA (2.68 µM) when compared to other combinations but statistically not significant.

Mycorrhizal association in gametophytes and sporophytes of the fern Pteris vittata (Pteridaceae) with Glomus intraradices

Ferns, which are usually colonizing different environments and their roots frequently present mycorrhization, have two adult stages in their life cycle, the sporophytic and the gametophytic phase. This paper describes the experimental mycorrhizal association between Pteris vittata leptosporangiate fern and a strain of Glomus intraradices during the life cycle of the fern, from spore germination to the development of a mature sporophyte. The aim of this study was to compare the colonization pattern of in vitro cultures of G. intraradices along the fern life cycle with those found in nature. For this, mature spores were obtained from fertile P. vittata fronds growing in walls of Buenos Aires city, Argentina. Roots were stained and observed under the light microscope for arbuscular mycorrhizal colonization. Approximately, 75 fern spores were cultured in each pot filled with a sterile substrate and G. intraradices (BAFC N° 51.331) as inoculum on the surface. After germination took place, samples were taken every 15 days until the fern cycle was completed. In order to determine colonization dynamics each sample was observed under optical and confocal microscope after staining. Gametophyte was classified as Adiantum type. Male and female gametangia were limited to the lower face, mycorrhizal colonization started when they were differentiated and took place through the rhizoids. Spores and vesicles were not found in this cycle stage. Paris-type mycorrhizal colonization was established in the midrib and in the embrionary foot. It was colonized by external mycelium. When the first root was developed soil inoculum colonized de novo this structure and Arum-type colonization was observed. This study proves that the type of colonization is determined by the structure of the host, not by the fungus. Both the gametophyte and embryo foot have determined growth and Paris-type colonization, while, sporophyte roots have undetermined growth and Arum-type colonization. The structures found in vitro cultures were highly similar to those found under natural conditions. Rev. Biol. Trop. 60 (2): 857-865. Epub 2012 June 01.

Los helechos presentan dos etapas en su ciclo de vida, una fase esporofítica y una gametofítica. Estos por lo general pueden colonizar diferentes ambientes y frecuentemente presentan raíces micorrizadas. Este estudio describe la asociación experimental entre Pteris vittata, un helecho leptosporangiado y una cepa de Glomus intraradices durante el ciclo de vida del helecho, desde la germinación de las esporas hasta el desarrollo del esporofito maduro. El objetivo de este estudio fue comparar los patrones de colonización de G. intraradices a lo largo de todo el ciclo de vida del helecho con los tipos encontrados en la naturaleza. Las esporas maduras fueron obtenidas de frondes fértiles de P. vittata que crecen sobre las paredes de la ciudad de Buenos Aires, Argentina. Las raíces se tiñeron y fueron observadas bajo microscopio óptico para el estudio de la colonización micorrízica. Aproximadamente 75 esporas de helecho se cultivaron en macetas con un sustrato estéril y con un inóculo de G. intraradices (N° 51.331 BAFC) en la superficie. Después de la germinación, se tomaron muestras cada 15 días hasta que se completó el ciclo de vida del helecho. Con el fin de determinar la dinámica de la colonización, cada muestra se observó con el microscopio óptico y el microscopio de confocal luego de la tinción correspondiente. El gametofito fue clasificado como del tipo “Adiantum”. Los gametangios femeninos y masculinos se desarrollaron en la cara inferior del mismo. La micorrización comenzó cuando los gametangios estaban ya diferenciados y la colonización se produjo a través de los rizoides. Las esporas y las vesículas no se encontraron en esta fase del ciclo. La micorrizacion tipo Paris se observó sobre la línea de la nervadura central. El pie del esporofito fue colonizado por el micelio externo. Cuando la raíz se desarrolló, se colonizó “de novo”, y se observó una colonización de tipo Arum. Este estudio demuestra que el tipo de colonización está determinado por la estructura del helecho y no por el hongo. Tanto el gametofito como el pie del embrión tienen crecimiento definido y colonización tipo Paris, mientras que las raíces del esporofito presentan un crecimiento indeterminado y una colonización tipo Arum. Las estructuras que se encontraron bajo cultivo coinciden con las que se encontraron en condiciones naturales.

Preliminary antibacterial evaluation of fronds of Pteris quadriaurita Retz. towards bacteria involved in dermatological diseases.

Pteris quadriaurita, a medicinally relevant pteridophyte used as an antihelmintic plant in traditional systems of medicine. In the present study fronds of Pteris quadriaurita was evaluated for its antibacterial potential and phytochemical contents in various solvent extracts of the plant in increasing polarity towards pathogenic bacterial species involved in skin diseases in human being. Antibacterial activity was evaluated by disc diffusion method. The results indicated that fronds of the plant showed antibacterial activity especially in methanolic extract. The mathanolic extract of the plant showed maximum activity towards Pseodomonas aeruginosa, a multi-drug resistant strain. Petroleum ether and water extracts did not show any antibacterial activity towards any of the tested organisms. The occurrence of flavonoid and terpenoids content in the plants may be one of the reasons for their antibacterial activity. Methanolic extract of the plant exhibited minimum inhibitory concentration as 25mg/ml and minimum bactericidal concentration as 50mg/ml towards Pseudomonas aerogenosa.

Gametófitos y esporófitos jóvenes de cuatro especies de helechos del género Pteris (Pteridaceae) naturalizadas en América

Gametophytes and young sporophytes of four species of the fern genus Pteris (Pteridaceae) naturalized in the American continent. The pantropical fern genus Pteris L. has about 250 species of which 60 occur in the American continent. We studied the morphogenesis of the gametophyte, and the morphology of the young sporophytes of four species: P. cretica, P. ensiformis, P. multifida and P.vittata, together with a palynological analysis that includes the ability of spores to germinate. Gametophytes were obtained trough in vitro culture techniques with agar-gellified Knudson medium. Young sporophytes were placed in earth-sand (3:1) sterile substrate. We used light and SEM microscopy. Triletes spores predominate, but monolete, tetralete, and other types of apertura are often found. The viability of spores is not affected by the variation, so the term spore polymorphism is applied to the condition occurring among these species. Spore polymorphism is similar in P. cretica and P. multifida. Germination occurs following the Vittaria type, 3-7 days after the sowing. Filamentous, 3-5 celled gametophytes were found in P. cretica, P. multifida and P. vittata, and 7-9 celled in P. ensiformis. Development of gametophytes takes place following Adiantum type and eratopteris type. The symmetry of the laminae differ in gametophytes, those of P. ensiformis and P. multifida are similar and differ from the other two species, P. cretica and P. vittata. Gametophytes of P. ensiformis, P. multifida and P. vittata are bisexual and protandric, while male gametophytes were found in P. cretica. Antheridia correspond to the common leptosporangiate type; they are cylindric in P. vittata and ovoid in the other three species. Archegonia necks have 4 rows of 4 cells each. The sporophytes complete their development 3 months after sowing, and have indument close to the adult plants. P. cretica shows obligated apogamy. Rev. Biol. Trop. 58 (1): 89-102. Epub 2010 March 01.

El género pantropical Pteris L. tiene 250 especies de la cuales 60 están en el continente Americano. Se estudió la morfogénesis de los gametófitos, y la morfología de los esporófitos jóvenes de cuatro especies: P. cretica, P. ensiformis, P. multifida y P.vittata, junto con un análisis palinológico que incluye la capacidad de las esporas de germinar. Los gametófitos se obtuvieron mediante técnicas de cultivo in vitro. Los esporófitos jóvenes se trasladaron a sustrato estéril de tierra y arena (3:1). Se usó el microscopio de luz y el de barrido (SEM). Se encontraron esporas con diferentes tipos de aperturas. La germinación ocurre entre 3-7 días y corresponde al tipo Vittaria. Se encontraron gametófitos filamentosos formados por 3-5 células en P. cretica, P. multifida y P. vittata y por 7-9 células en P. ensiformis. El desarrollo gametofítico ocurre de dos formas: tipo Adiantum y tipo Ceratopteris. Los gametófitos de P. ensiformis, P. multifida y P. vittata son monoicos y protándricos. P. cretica desarrolla gametófitos anteridiados. Los anteridios corresponden al tipo común de los helechos leptosporangiados, son cilíndricos en P. vittata y ovoides en las otras tres especies. Los cuellos de los arquegonios tienen 4 hileras con 4 células cada una. Los esporófitos se desarrollan después de los 3 meses de su siembra y su indumento es semejante a las plantas adultas. P. cretica presenta apogamia obligada.

Una especie nueva de helecho del género Pteris (Pteridaceae) endémica de Costa Rica / A new species of fern of the genus Pteris (Filicales: Pteridaceae) endemic to Costa Rica

The new fern species Pteris herrerae A. Rojas & M. Palacios, endemic to Costa Rica, is described. It differs from P. decurrens C. Presl in basal segments reduced to 1/5-1/2 of the next segment (vs. 2/3-3/4), basal pinnae not bifurcated (vs. bifurcated), pinnae apex mucronate (vs. acuminate) and segment apex undulate (vs. dentate). It differs from Pteris consanguinea in the elliptic pinnae (vs. oblong), two segments reduced on the base (vs. lack), segments entire to undulate (vs. dentate), basal pinnae without basiscopic lobes (vs. with basiscopic lobes) and segment apex entire to undulate (vs. dentate).

Se describe Pteris herrerae A. Rojas & M. Palacios, endémica de Costa Rica. Esta es diferente de P. decurrens C. Presl por segmentos basales reducidos a 1/5-1/2 del tamaño de los siguientes (vs. 2/3-3/4), pinnas basales no bifurcadas (vs. bifurcadas), ápice de las pinnas mucronado (vs. acuminado) y ápice de los segmentos ondulado (vs. dentado). También es diferente de Pteris consanguinea Mett. ex Kuhn por pinnas deltado-lanceoladas (vs. oblongas), con un par de segmentos reducidos en la base (vs. sin ellos), pinnas basales sin lóbulos basicópicos alargados (vs. con lóbulos basiscópicos) y segmentos enteros a ondulados (vs. dentados).

Mechanisms of inhibition on human pancreatic cancer PC-3 cell line by a diterpenoid compound 5 F isolated from Pteris semi pinnata L treatment / 中华中医药杂志

Objective:To investigate the effect of 5F on proliferation and apoptosis of human pancreatic cancer.Methods:The pcDNA3.1-PUMAAS and pcDNA3.1 were transfected into AsPC-1 cells by Lipofectamine 2000 methods,stable transfected clony was chosen through G418.AsPC-1 cells were divided into three groups:transfected pcDNA3.1-PUMAAS,transfected pcDNA3.1 and control group without transfection AsPC-1 cells.Three groups was treated with serial concentrations(8.875,37.5,142?mol/L,respectively)of 5F 10?L.MTT assay was used to observe the inhibitory actions of 5F on AsPC-1 cells.The apoptotic rate of the cells was detected by flow cytometry.And the apoptosis was assessed by Hoechst 33258 dye staining and TUNEL staining.The expression of PUMA protein was detected by Western blotting and RT-PCR.Results:The inhibitory action on cell growth was seen in AsPC-1 control cells and pcDNA3.1-cells dealing with 5F.It could also promote the occurrence of apoptosis.5F inhibited the proliferation of AsPC-1 cells in a concentration-dependent manner.The apoptosis induced by 5F was accompanied with the up-regulation of PUMA.In cells treated with 5F,the apoptosis rate decreased greatly and proliferation rate increased compared with the AsPC-1 control cells and pcDNA3.1-cells dealing with 5F.The expression of PUMA was decreased greatly comparing with pcDNA3.1-cells treated with 5F,Conclusion:5F can depress the proliferation of AsPC-1 cell in vitro,mainly through the induction of apoptosis,and it was a potential agent for pancreatic cancer chemotherapy,the mechanism was probably related to its effect on the regulation of PUMA expression.

Effects of Extract 5F from Pteris Semipinnata L. on the Apoptosis of Skin Fibroblasts in Patients with Systemic Scleroderma / 中华皮肤科杂志

Objective To study the effects of extract 5F from Pteris semipinnata L. on the apoptosis of skin fibroblasts in patients with systemic sc leroderma (SSc) and the possible mechanism of its inhibition on the proliferatio n of skin fibroblasts. Methods The cultured SSc skin fibroblasts were used as an experimental model, and the apoptosis of SSc skin fibroblasts before and afte r the extract 5F stimulation was detected by TUNEL assay and fluorescent microsc opy. Results The apoptosis of SSc skin fibroblasts was induced by the extract 5F, and the apoptosis was enhanced along with the increase of the concentration and incubation time of this compound within definite limits. Conclusions These results indicate that the extract 5F can induce the apoptosis of SSc skin fibro blasts, and significantly inhibit the proliferation and collagen synthesis in th ese cells.

Purification of 5F from Pteris semipinnata and its enhanced cytotoxicity in vitro / 中国药理学通报

AIM To find a new method for purifying the active compound 5F isolated from Pteris semipinnata L.(PsL ) and observe its enhanced cytotoxicity when combined with other antitumor agents. METHODS Silica gel combined with AgNO 3 was made to purify 5F. Cytotoxicity was detected with trypan blue dye exclusion assay. RESULTS After purification, the concentration of compound 5F in purified products was higher than 99%. The inhibitory rates of 5F combined with 5 Fu or CDDP or VCR were higher than that of these drugs used alone. CONCLUSIONS Compound 5F could be purified effectively using silica gel combined with AgNO 3. 5F could enhance the cytotoxicity on HL 60 and K562 cells of the drugs mentioned above. The different effect of these agents on the various phases of cell cycle kinetics may explain the enhanced effect of 5F.

The effects of a diterpenoid compound 5F isolated from pteris semipinnata L on the activity and expression of mitogen activated protein kinase in K562 cells / 中国药理学通报

AIM: To study the effects of a diterpenoid compound 5F (ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic acid) isolated from pteris semipinnata L on the the activity and expression of mitogen activated protein kinase of K562 cells. METHODS: K562 cells were treated with compound 5F for 24 hours, then MAPK activity was determined using MBP as substrate, Western blotting was used to detect the effect of compound 5F on the expression of MAPK. RESULTS: The activity and expression of MAPK increased after the treatment of 5F. CONCLUSION: One mechanism of the anti-tumor effect of compound 5F is through abnormal activation and abnormal expression of MAPK.

Application of AgNO3-Processed Silica Gel Chromatography in Isolation and Purification of Active Components of Herbal Medicine / 广州中医药大学学报

[Objectivs] To investigate the principle of AgNO3-processed silica gel chromatography in isolation and purification of mixtures with carbon-carbon double bonds (CCDB) from herbal medicine. [Methods] AgN03-processed gel chromatography was used for the isolation of mixtures with carbon-carbon double bonds in the volatile oils and active components from Pteris semipinnata L. ; silver-ion complexion chromatography was applied to isolate arachidom'c acid from lipids. The principle of AgNO3-processed silica gel chromatography was analyzed by coordination theory. [Results] Stable AgNO3 coordinate complexes were obtained. [ Conclusion ] The principle of the formation of stable AgNO3 coordinate complexes may be: (1) the stability will increase with CCDB in free space; (2) exocyclic CCDB is more stable than that of intra-cyclic CCDB; (3) cis-form CCDB is more stable than that of trans-form CCDB; (4) terminal CCDB is more stable than that of cis-form CCDB; (5) aromatic hydrocarbon with multiple benzene rings is more stable than that with single ber zene ring. The possible principle will benefit to the isolation and purification of the mixtures with CCDB, which have similar structure and physiochemical property and are difficult to be isolated and purified by common chromatography and extriction method.

Role of calcium ion in the cyotoxicity and DNA fragment induction in HL-60 cells by 6F isolated from Pteris semipinnata L / 中国病理生理杂志

AIM: To investigate the changes of cytosolic free calcium concentration ([Ca 2+ ] i) and expression of Bcl-2 in HL-60 cells treated by 6F isolated from Pteris semipinnata L. (PSL), and to discuss the relations between calcium ion and cytotoxicity and DNA fragment induction effects of 6F. METHODS: HL-60 cells were used as in vitro model. [Ca 2+ ] i was measured on fluorescent spectrophotometry using Fura-2/AM as Ca 2+ indicator. Bcl-2 expressing level was measured by flow cytometry. Tetrazolium salt (MTT) and diphenylamine staining methods were applied for cytotoxicity assay and DNA fragmentation detection, respectively. RESULTS: [Ca 2+ ] i increased obviously in a dose and time dependent manner after treated HL-60 cells with 6F. 6F decreased the expressing level of Bcl-2. Adding 2 mmol/L Ca 2+ to the medium, or 1 mmol/L EDTA to chelate Ca 2+ , or 4 ?mol/L calcium ionophore A 23187 to increase the concentration of cytosolic Ca 2+ , the DNA fragment induction by 6F was not affected, whereas the cytotoxicity of 6F was enhanced. 250 ?mol/L Zn 2+ attenuated the DNA fragment induction, and the cytotoxicity of 6F against HL-60 cells was enhanced significantly. CONCLUSION: It was speculated that the decreased expressing of Bcl-2 by compound 6F was related to increased [Ca 2+ ] i in HL-60 cells, and DNA fragment induction was possibly catalyzed by Ca 2+ - independent DNase.