One new cyclopeptide was isolated from the ethyl acetate fraction of the 75% EtOH extract of Selaginella tamariscina by various column chromatography methods(HP-20, polyamide and semi-preparative HPLC). Its structure was identified as selapeptin A(1) by extensive spectroscopic analysis(HR-ESI-MS, 1 D and 2 D NMR). Compound 1 was evaluated for cytotoxic activities by MTT assay. It showed potent cytotoxic activity against B16 F10 with the inhibition rate of 51.57%±4.34% at 40 μmol·L~(-1) while had no impacts on MDA-MB-231 and MDA-MB-468 at 100 μmol·L~(-1).
Chromatography, High Pressure Liquid ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Peptides, Cyclic/pharmacology* ; Selaginellaceae/chemistry*

Abstract: A new selaginellin derivative named as selaginellin S (1) was isolated from the whole plants of Selaginella pulvinata (Hook. et Grev.) Maxim. (Selaginellaceae), together with a known one (selaginellin M, 2). Compounds 1 and 2 were separated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were determined on the basis of extensive spectroscopic analysis including IR, MS, 1D and 2D NMR experiments, as well as ECD calculations. Compound 1 is a key intermidiant in the biosynthesis pathway of selaginellins. Compound 2 is first reported in this plant.
Biphenyl Compounds ; chemistry ; isolation & purification ; Cyclohexanones ; chemistry ; isolation & purification ; Molecular Structure ; Selaginellaceae ; chemistry

In the current study, nine flavonoids were isolated and purified from 75% ethanol extract of Selaginella uncinata (Desv.) Spring by column chromatographic techniques over macroporous resin, polyamide, silica gel, Sephadex LH-20 and pre-HPLC. On the basis of their physico-chemical properties and spectroscopic data analyses, these compounds were elucidated as cirsimarin (1), nepitrin (2), apigenin-6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (3), apigenin-6-C-β-D-glucopyranosyl-8-C-α-L-arabinopyranoside (4), apigenin-7-O-β-D-glucopyranoside (5), 2,3-dihydroamentoflavone (6), 4'-O-methylamentoflavone (7), 2,3-dihydro-4'-O-methyl-amentoflavone (8), and 2,3,2",3"-tetrahydron-4'-O-methyl-robustaflavone (9). Compounds 1-5 belong to flavonoid glycosides and were isolated from the genus Selaginella for the first time.
Flavonoids ; analysis ; Selaginellaceae ; chemistry

AIM: To investigate the chemical constituents of Selaginella sinensis (Desv.) Spring. METHODS: Chromatographic separations on Diaion HP-20, silica gel, and Sephadex LH-20 were used. The structures of the isolates were elucidated on the basis of spectroscopic analysis, as well as chemical methods. RESULTS: Eight compounds were obtained and their structures were identified as sinensioside A (1), syringaresinol-4- O-β-D-glucopyranoside (2), (+)-medioresinol-4-O-β-D-glucopyranoside (3), pinoresinol-4, 4'-di-O-β-D-glucopyranoside (4), quercetin (5), eucomic acid (6), shikimic acid (7), and 2, 3-dihydroamentoflavone (8). CONCLUSION Compound 1 is a new dihydrobenzofuran sesquilignan glycoside from Selaginella sinensis.
Benzofurans ; chemistry ; isolation & purification ; Furans ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Lignans ; chemistry ; isolation & purification ; Molecular Structure ; Plant Extracts ; chemistry ; Plant Stems ; chemistry ; Quercetin ; chemistry ; isolation & purification ; Selaginellaceae ; chemistry

This study is to observe the protection effect of amentoflavone (AMT) in Selaginella tamariscina against TNF-alpha-induced vascular inflammation injury of endothelial cells. On the basis of TNF-alpha induced human umbilical vein endothelial cell, observe the influence of AMT on endothelial active factor, the contents of SOD and MDA, the protein expression of vascular endothelial adhesion molecules and inflammatory factor; study the effect of its common related signal pathways such as NF-kappaB; research the effect of AMT against TNF-a induced human umbilical vein endothelial cell injury by means of MTT, ELISA, Western blotting and the cell immunofluorescence. The results showed that AMT could increase the content of NO and decrease the levels of VCAM-1, E-selectin, IL-6, IL-8 and ET-1; enhance the activity of SOD, reduce the content of MDA; downregulate the protein expressions of VCAM-1, E-selectin, NF-kappaBp65 and up-regulate IkappaBalpha, attenuate the NF-kappaBp65 transfer to cell nucleus. AMT has the effect of protect vascular endothelial and maybe via the signal pathway of NF-kappaB to down-regulate the inflammation factor and oxidative damage factor of downstream.
Biflavonoids ; isolation & purification ; pharmacology ; Cell Cycle ; drug effects ; Cell Survival ; drug effects ; E-Selectin ; metabolism ; Endothelin-1 ; metabolism ; Human Umbilical Vein Endothelial Cells ; Humans ; I-kappa B Proteins ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Malondialdehyde ; metabolism ; NF-KappaB Inhibitor alpha ; Nitric Oxide ; metabolism ; Plants, Medicinal ; chemistry ; Protective Agents ; isolation & purification ; pharmacology ; Selaginellaceae ; chemistry ; Superoxide Dismutase ; metabolism ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; adverse effects ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo establish a HPLC-DAD model for the simultaneous determination of two selaginellins (selaginellin and selaginellin B) and four biflavones (amentoflavone, sequoiaflavone, hinokiflavone and isocryptomerin) contained in 10 batches of Selaginella tamariscina and 12 batches of S. pulvinata produced in different areas.

METHODThe analysis was performed on a Waters Cosmosil C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitril-0.1% formic acid as mobile phase in a linear gradient mode. The detection wavelength was set at 280, 337 nm. The flow rate was kept at 1.0 mL x min(-1), and the column temperature was 30 degrees C.

RESULTThe six active constituents showed significant different in content. Amentoflavone in S. tamariscina contains (5. 628-9. 184 mg x g(-1)) is more than that contained in S. pulvinata (0.823-7.131 mg x g(-1)), while selaginellin in S. pulvinata (0.123-0.593 mg x g(-1)) is more than that contained in S. tamariscina (0.067-0.133 mg x g(-1)). All the calibration curves showed good linear correlation coefficients (r > 0.9997) over the wide test ranges.

CONCLUSIONThe developed HPLC-DAD method is simple, sensitive and accurate and has the good repeatability in separation, which is available for the quality control of S. tamariscina and S. pulvinata.

Biflavonoids ; chemistry ; Biphenyl Compounds ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Cyclohexanones ; chemistry ; Flavones ; chemistry ; Flavonoids ; chemistry ; Selaginellaceae ; chemistry

Plant dehydroascorbate reductase (DHAR) is a physiologically important reducing enzyme in the ascorbate-glutathione recycling reaction. In this study, two DHARs genes (SmDHAR1 and SmDHAR2) were isolated from Selaginella moellendorffii. The SmDHAR1 and SmDHAR2 genes encode two proteins of 218 and 241 amino acid residues, with a calculated molecular mass of 23.97 kDa and 27.33 kDa, respectively. The genomic sequence analysis showed SmDHAR1 and SmDHAR2 contained five and six introns, respectively. Reverse transcription PCR revealed that the SmDHAR1 and SmDHAR2 were constitutive expression genes in S. moellendorffii. The recombinant SmDHAR1 and SmDHAR2 proteins were overexpressed in E. coli, and were purified by Ni-affinity chromatography. The recombinant SmDHAR1 showed 116-fold higher enzymatic activity towards the substrate dehydroascorbate than recombinant SmDHAR2. The recombinant SmDHAR1 showed higher thermal stability than recombinant SmDHAR2. These results indicated obvious functional divergence between the duplicate genes SmDHAR1 and SmDHAR2.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; DNA, Plant ; genetics ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Oxidoreductases ; biosynthesis ; chemistry ; genetics ; Plant Proteins ; biosynthesis ; chemistry ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Selaginellaceae ; enzymology ; genetics ; Sequence Analysis, DNA

OBJECTIVETo establish a method to analyze HPLC fingerprint characteristics of 10 plants from Selaginella.

METHODHPLC was applied for establishment of fingerprints, which were used to evaluate and distinguish the different species of Selaginella.

RESULTThe different species from Selaginella showed different HPLC fingerprint characteristic. The samples of the same species but collected in different period, different environment or different locations showed certain difference in fingerprints

CONCLUSION2 important mutual fingerprint peaks were found in the 10 plants of Selaginella species and 5 peaks can be used as "main fingerprint peaks". The dates of these peaks can used for assessment of phylogenetic relation among species and evaluation of quality.

Biflavonoids ; analysis ; China ; Chromatography, High Pressure Liquid ; methods ; Plants, Medicinal ; chemistry ; classification ; Selaginellaceae ; chemistry ; classification ; Species Specificity

OBJECTIVETo observe the Chinese herbal medicine Selaginella-induced radiosensitization of terminal nasopharyngeal carcinoma (NPC).

METHODSTotally 180 patients with NPC were divided equally into 3 groups with the same radiotherapeutic protocols. The patients in group A received radiotherapy alone, those in group B were given daily Selaginella (30 g) prepared into 50 ml decoction during the entire course of radiotherapy, and those in group C had Selaginella 30 g daily in the late course of radiotherapy.

RESULTSThe complete remission rate of nasopharyngeal primary lesions in groups B and C was significantly higher than that in group A, with also significantly higher complete remission rates of the cervical lymph nodes. The acute toxicity of the skin and mucous membrane was milder in the latter two groups, but the differences were not significant.

CONCLUSIONSelaginella may induce radiosensitization for terminal NPC and does not increase the acute toxicity of radiotherapy.

Adult ; Carcinoma, Squamous Cell ; drug therapy ; radiotherapy ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; drug therapy ; radiotherapy ; Phytotherapy ; Radiation-Sensitizing Agents ; therapeutic use ; Selaginellaceae ; chemistry ; Treatment Outcome

OBJECTIVETo evaluate the estrogenic activity of several kinds of Chinese herbal medicines.

METHODUse zoopery and reporter gene technique to study the estrogenic activity of five Chinese herbal medicines. Zoopery: weanling female Kunming mice weight 9-12 g were administrated botanical extracts of Selaginella tamariscina, Pinus Massoniana, Corallodiscus flabellate, Dryopteris sublaeta and Leonurus heterophyllus, the positive control group with Nilestriol tablets and control group with water, respectiely. On the eighth day, the animals were sacrificed and the uteri were separated solely and weighed. Reporter gene technique: Induce the expression of reporter gene controlled by ERE and measure the activity of luciferase on cell's clear supernatant.

RESULTThe botanical extracts of S. tamariscina can increase weights of mice (P < 0.01); In the expression of reporter gene controlled by ERE, Either ERalpha or ERbeta's has estrogenic activity (P < 0.01). Follow in the zoopery we find the water part and the n-butanol part of S. tamariscina are the two active parts.

CONCLUSIONS. tamariscina and it's water part and n-butanol part have estrogenic activities, effect on ERbeta is greater than ERalpha.

Animals ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Estrogen Receptor alpha ; metabolism ; Estrogen Receptor beta ; metabolism ; Female ; Leonurus ; chemistry ; Mice ; Organ Size ; drug effects ; Phytoestrogens ; isolation & purification ; pharmacology ; Pinus ; chemistry ; Plants, Medicinal ; chemistry ; Selaginellaceae ; chemistry ; Uterus ; anatomy & histology