Stellera chamaejasme extract against multidrug resistance of breast cancer cell line MCF-7 / 中国中药杂志

This study explored the effect and underlying mechanism of Stellera chamaejasme extract(SCE) on multidrug resistance of breast cancer. The chemotherapy-sensitive breast cancer cell line MCF-7 and adriamycin(ADR)-resistant cell line MCF-7/ADR were used as experimental subjects. MTT assay was used to detect cell proliferation activity. Pi staining was used to detect the cell cycle. 4',6-Diamidino-2-phenylindole, dihydrochloride(DAPI) staining and flow cytometry were used to detect apoptosis. Dansylcadaverine(MDC) staining and GFP-LC3B-Mcherry adenovirus transfection were used to detect autophagy. The protein expression of Bcl-2, Bax, caspase-9, caspase-3, LC3B, p62, and Beclin-1 was detected by Western blot. The results showed that SCE could significantly inhibit the proliferation of both sensitive and resistant breast cancer cell lines. The drug resistance factor was 0.53, which was significantly lower than 59 of ADR. Meanwhile, the proportion of sensitive/resistant cells in the G_0/G_1 phase increased significantly after SCE treatment. In addition, DAPI staining showed that a series of apoptosis phenomena such as nuclear pyknosis, staining deepening, and nuclear fragmentation appeared in sensitive/resistant cell lines after SCE administration. Moreover, the results of flow cytometry double staining showed that the proportion of apoptotic cells in sensitive/resistant cell lines increased significantly after SCE administration. Besides, Western blot showed that the protein expression levels of caspase-3, caspase-9, and Bcl-2 significantly decreased and the expression level of Bax protein significantly increased in both breast cancer cell lines after SCE administration. Furthermore, SCE could also increase the positive fluorescent spots after MDC staining and yellow fluorescent spots after GFP-LC3B-mcherry transfection, and up-regulate the expression levels of autophagy-related proteins LC3B-Ⅱ, p62, and Beclin-1 in breast cancer cells. In summary, SCE may play the role of anti-multidrug resistance by blocking the cell cycle of breast cancer multidrug-resistant cells, blocking autophagy flow, and ultimately interfering with the apoptosis resistance of drug-resistant cells.

Chemical constitutents from flowers of Stellera chamaejasme and their antioxidant activity / 中草药

Objective: To investigate the flavonoids and lignans from the flowers of Stellera chamaejasme and their structure-activity relationship (SAR) of antioxidant activity. Methods: The compounds were isolated by column chromatography and HPLC packed with macroporous resin, silica gel, and Sephadex LH-20. Their structures were elucidated by spectroscopic analysis. Their anti-oxidant activities in vitro were evaluated by DPPH, ABTS, and FRAP assays. Results: Twelve compounds were isolated from the flowers of S. chamaejasme, and identified as artemisetin (1), quercetin (2), isoscutellarein-8-O-β-D-glucuronopyranoside (3), quercetin-3-O-β-D- glucopyranoside (4), astragalin (5), hypolaetin-8-O-β-D-glucuronopyranoside (6), kaempferol 3-O-β-D-glucopyranosyl-(1→2)-O-α- L-xylopyranoside (7), rel-(3R,3'S,4R,4'S)-3,3',4,4'-tetrahydro-6,6'-dimethoxy [3,3'-bi-2H-benzopyran]-4,4'-diol (8), matairesinol (9), uralenol (10), cycloastragenol (11), and (+)-pinoresinol (12). Conclusion: Compounds 1, 3, 5-7, and 10 are isolated from this plant for the first time, and compounds 2, 4, 5, and 10 showed significant antioxidant activity, and the SAR analysis suggested that the glycosylation at the C-8 or C-3 position of flavonoids could weaken their antioxidant activity.

Chemical Constituents from Stellera chamaejasme Linn / 中国药学杂志

OBJECTIVE: To study the chemical constituents from the roots of Stellera chamaejasme Linn. METHODS: The 95% ethanol extract of the roots of Stellera chamaejasme Linn. was fractionalized by using petroleum ether, dichloromethane and ethyl acetate, respectively. The extraction parts were isolated by various chromatographic methods including silica gel, ODS column and preparative HPLC methods. Their structures were elucidated on the basis of spectroscopic data such as HR-ESI-MS and NMR. RESULTS: Seventeen compounds were isolated and identified as follows: naringenin (1), (-)-epiafzelechin (2), (+)-epiafzelechin (3), 2,6,2',6'-tetramethoxy-4,4'-bis (2,3-epoxy-1-hydroxypropyl) biphenyl (4), 8-hydroxypluviatolide (5), (-)-syringaresinol (6), prestegane B (7), 7-hydroxycoumarin (8), daphnoretin (9), 3-(2,4-dihydroxyphenyl) propanoic acid methyl ester (10), 3,4-dihydroxystyryl alcohol (11), 3-[(1E)-3-hydroxy-1-propen-1-yl]-2, 5-dimethoxyphenyl-β-D-glucopyranoside(12), Z-octadecyl caffeate (13), 1-glyceryl linolenate (14), glycerol monolinoleate (15), 2, 3-dihydroxypropyl-9-octadecenoate (16) and β-sitosterol (17). CONCLUSION: Compounds 1, 2, 4, 5, 9-15 are reported from the genus Stellera Linn. for the first time.

A new guaiane sesquiterpene from rhizome of Stellera chamaejasme / 中草药

Objective To study the chemical constituents from rhizome of Stellera chamaejasme. Methods The compound was isolated and purified by various chromatographic techniques, and its structure was elucidated by spectroscopic analysis. Results A new sesquiterpene was obtained from 95% ethanol extract of rhizome of S. chamaejasme and identified as 1β,10β-dihydroxyguaia- 4,11-dien-3-one (1). Conclusion Compound 1 is a new guaiane sesquiterpene and named stelleraguaianone A. Meanwhile, the compound with a guaiane sesquiterpene skeleton is also obtained from the genus Stellera for the first time.

A new lignan from rhizome of Stellera chamaejasme / 中国中药杂志

To investigate the chemical compounds from the rhizome of Stellera chamaejasme, nine lignans, including stellerachamin A (1), 8-hydroxypluviatolide (2), wikstromol (3), pinoresinol (4), matairesinol (5), dextrobursehernin (6), hinokinin(7), (-)-glaberide I (8) and （-） medioresinol (9) were isolated by various chromatographic methods. Their structures were extensively determined on basis of MS and NMR spectroscopic data analysis. Among them, compound 1 was a new lignan, and compounds 2 and 7 were isolated from Thymelaeaceae for the first time.

Transcriptome characterization of Stellera chamaejasme with Illumina sequencing technology / 中草药

Objective To obtain the transcriptome database and gene sequence, SSR as well as transposon information of Stellera chamaejasme. Methods Using the high-throughput sequencing platform (Illumina HiSeq 2000), a root transcriptome dataset of S. chamaejasme was obtained, and the sequencing results were analyzed with the bioinformatic way. Results With a total of 26 785 872 clean reads, 47 053 unigenes were assembled. All these unigenes were then blasted with Nr, Swiss-Prot, KEGG, COG, and GO databases. There were 11 138 and 24 744 unigenes were annotated with Nr and Swiss-Prot databases, respectively. The unigenes were involved in 36 GO-terms and 119 metabolic pathways. Further analysis showed that 15 unigenes were involved in terpenoids biosynthesis. Using MISA software, the results showed that there were 3 480 SSR from the 47 053 unigenes, and the most type of SSR was mononucleotide (1 986) with the frequency of 57.07%. Moreover, the hexanucleotide only had five repeat SSR and the frequency was only 0.14%. With RepeatMasker online tools to analyze the transposon of the transcriptome sequences, the results indicated that there were 1 497 transposons, and the number of transposons with E < 1×10-5 was 827. All the transposons were grouped into 22 types, and the LINE/L1 type (405) had the highest frequency (48.97%). The DNA/Ginger, DNA/hAT, DNA/PIF-ISL2EU, and LINE/Jockey as well as LTR/Lenti were the least type since each of them has only one transposon. Conclusion In this study, rich sequence information of gene, SSR as well as transposon information of Stellera chamaejasme is helpful to carry out the research of the molecular mechanism of phorbol ester biosynthesis in S. chamaejasme in the future.

A new biflavone glucoside from the roots of Stellera chamaejasme / 中国天然药物

The present study investigated the chemical constituents of the roots of Stellera chamaejasme (Thymelaeaceae). One new biflavone glucoside (1), along with other thirteen known compounds (2-14), was isolated by repeated column chromatographic methods and their structures were elucidated on the basis of spectral analyses. The cytotoxic activities of selected compounds were evaluated against four human cancer cell lines (A549, BEL-7402, HCT-116, and MDA-MB-231) by the SRB assay method. Compound 9 showed remarkable cytotoxicity against BEL-7402 with IC50 value being 0.65 μg·mL(-1); compounds 7, 8, and 12 exhibited significant cytotoxic activity against A549 with IC50 values being 2.38, 1.57, and 2.35 μg·mL(-1), respectively.

Isochamaejasmin induces apoptosis in leukemia cells through inhibiting Bcl-2 family proteins / 中国天然药物

The biflavonoid isochamaejasmin is mainly distributed in the root of Stellera chamaejasme L. (Thymelaeaceae) that is used in traditional Chinese medicine (TCM) to treat tumors, tuberculosis, and psoriasis. Herein, isochamaejasmin was found to show similar bioactivity against Bcl-2 family proteins to the reference Bcl-2 ligand (-)-gossypol through 3D similarity search. It selectively bound to Bcl-xl and Mcl-1 with Ki values being 1.93 ± 0.13 μmol·L(-1) and 9.98 ± 0.21 μmol·L(-1), respectively. In addition, isochamaejasmin showed slight growth inhibitory activity against HL-60 with IC50 value being 50.40 ± 1.21 μmol·L(-1) and moderate growth inhibitory activity against K562 cells with IC50 value being 24.51 ± 1.62 μmol·L(-1). Furthermore, isochamaejasmin induced apoptosis of K562 cells by increasing the intracellular expression levels of proteins of the cleavage of caspase-9, caspase-3, and PARP which involved in the Bcl-2-induced apoptosis pathway. These results indicated that isochamaejasmin induces apoptosis in leukemia cells by inhibiting the activity of Bcl-2 family proteins, providing evidence for further studying the underlying anti-cancer mechanism of S. chamaejasme L.

Inhibition of Extract from Leaf of Stellera chamaijasme L on Dermatic Staphylococcus aureus and Trichophyton gypseum / 医药导报

Objective To study the antibiotic activities of leaf extract of Stellera chamaejasme L on the skin. Methods Activities of the leaf extract on Trichophyton gypseum and Staphylococcus aureus was examined by antibiotic susceptibility paper disk method and tube double dilution method. Results When extracted by 95%ethanol,the extract from Stellera chamaejasme L at 100 mg·mL-1 formed an inhibition zone with the diameter of 12. 8 mm for Staphylococcus aureus and 12. 0 mm for Trichophyton gypseum. When extracted by dichloromethane,the minimum inhibitory concentration( MIC)of the extract from Stellera chamaejasme L was 0. 25 mg·mL-1 for Staphylococcus aureus and 0. 50 mg·mL-1 for Trichophyton gypsem. Conclusion Leaf extract of Stellera chamaejasme L has an antibacterial activity against Staphylococcus aureus and Trichophyton gypseum and the antibacterial activity against Staphylococcus aureus is stronger.

Accumulation of flavonoids and coumarins during adventitious bud growth in Stellera chamaejasme / 中草药

Objective: To investigate the accumulation of flavonoids and coumarins during the growth of adventitious buds in Stellera chamaejasme. Methods: The optimal culture media for the induction and the growth of adventitious buds were screened through tissue culturing. The dynamic accumulation of the total flavonoids and three components of coumarin (umbelliferone, daphnetin, and scopoletin) was analyzed by visible spectrophotometry and high-performance liquid chromatography methods, respectively. Results: The optimal condition for the induction of adventitious bud proliferation was MS + 0.5 mg/L BA + 0.5 mg/L NAA; After 4-week culture, the number of adventitious buds in every tip-bud was 35; The optimal condition for the growth of adventitious bud was MS + 0.3 mg/L BA + 0.1 mg/L NAA; After 4-week culture, the average length of buds was 2.3 cm; The amount of total flavonoids increased to the highest of 0.218 g after 5-week culture of the adventitious buds; The amount of the three components of coumarin increased with culturing time increasing during the growth of adventitious buds, and increase to the highest at week 5. But the contents of the components were lower than those in the wild roots, and the descending order had changed. Conclusion: The accumulation of flavonoids and coumarins in adventitious buds of S. chamaejasme could reach to the highest at week 5 of culture.

Effects of four extracts of Chinese herbal medicines on cellular immunity in rats induced by multiple drugs resistant bacillus tuberculosis among pneumoconiosis patients complicated with tuberculosis / 中华微生物学和免疫学杂志

Objective To investigate the regulative effect of the extract of Radix Ranunculi Ternati,Radix Sophorae Flavescenti,Prunella vulgaris L.and Stellera chamaejasme L.on cellular immunity induced by multiple drugs resistant bacillus tuberculosis(MDR-TB) from pneumoconiosis patients complicated with tuberculosis in rats.Methods MDR-TB model in rats was induced by MDR-TB.Normal control group were feed by standard feed,model group were irrigated by normal saline,and the other groups were respectively feed by the extract of Chinese herbal medicines.The content of IFN-γ,IL-4,IL-10 and IL-12 were examined by ELISA.RT-PCR was used to detect the mRNA levels of them.Results The content of IFN-γof the four extracts of Chinese herbal medicines were (2.01 ±0.73 ),( 1.92±0.56),( 1.98 ±0.67 ) and (1.94±0.59) pg/ml,IL-4 were (6.01±1.46),(6.12±1.35),(6.47±1.46) and (6.15±1.44) pg/ml,IL-10 were (12.09±3.07),( 12.45±4.01 ),( 12.13±3.43) and (12.54±3.78) pg/ml,IL-12 were (2.99±0.89),(2.75±0.84),(3.02±0.86) and (2.89±0.75) pg/ml.Compared to the model group,they resulted in significant in serum IFN-γ,IL-12,IL-4 and IL-10 (P＜0.05 or P＜0.01 ),the mRNA levels of them were significantly (P＜0.05 or P＜0.01 ).Conclusion The four extracts of Chinese herbal medicines can enhance the cellar immunological function in rats from up-regulation of the level of genetic transcription,accordingly provide the theory base of healing of pneumoconiosis patients complicated with tuberculosis with them.

Effects of Serum of Mice Treated With Ruixiang Langdu(Stellera chamaejasme L.) abstract on Proliferation and Clonal Formation of K562 Cells / 中国药房

OBJECTIVE:To study the antitumor mechanism of Ruixiang Langdu(Stellera chamaejasme L ) abstracts(SCA) METHODS:SCA-contained serum was derived from mice pre-administrated with different oral dosages of SCA and at different times after administration The effects of the serum on the proliferation of K562 leukemic cells were observed with MTT assay and clone formation RESULTS:After mixing with the SCA-contained serum derived at 1,2,4,8h after giving SCA(3,6 and 12g/kg),the rates of MTT transformation and clone formation of K562 cells were decreased significantly The SCA-contained serum 12 h after giving drug was more effective than others CONCLUSION:The SCA-contained serum inhibited the proliferation of tumor cells,which may be one of its important antitumor mechanism

Effect of serum Ruixiang Langdu (Stellera chamaejasme) extract on proliferation, clonal formation and DNA synthesis of mouse L_(1210) leukemic cells / 中草药

Object To explore, on the whole, the anticancer activity and mechanism of the aqueous extract of Ruixiang Langdu (Stellera chamaejasme L.) (SCLA). Methods Mice were given different i.g. doses of SCLA and their serum collected at different intervals after drug administration. The effects of the serum on the proliferation of mouse L 1210 leukemic cells were observed with MTT assay, clone formation and incorporation of [ 3H] TdR into DNA of the cells. Results Serum SCLA collected 1, 2, 4, 8 h after administration of 3, 6, and 12 g/kg SCLA showed significant decrease of MTT formazans and clone formation. Serum SCLA collected 2 h after drug administration showed more strong inhibition of cancer cell proliferation. It also inhibited the incorporation of [ 3H] TdR into DNA of L 1210 cells. Conclusion SCLA showed a remarkable inhibitory effect on proliferation and DNA synthesis of cancer cells cultured in vitro, which may be the main anticancer mechanisms of SCLA.

Chemical constituents from roots of Stellera chamaejasme / 中草药

Object To study the chemical constituents from the roots of Stellera chamaejasme L. Methods Chemical constituents were isolated by chromatographic methods and identified by physiochemical and spectral analysis. Results One lignan and three biflavonoids were purified and determined as bursehernin (Ⅰ), chamaejasmenin B (Ⅱ), isoneochamaejasmin A (Ⅲ),and (+)-chamaejasmin (Ⅳ). Conclusion Compound Ⅲ and Ⅳ are determined as new optical compounds. Compound I is first isolated from Stellera L. species.

Stellera chamaejasme induced apoptosis of HL-60 cells and regulated expression of bcl-2 protein in SGC-7901 cells / 中草药

Object To explore the antitumor mechanism of Stellera chamaejasme Linn. (SC). Methods SC containing-serum (SCCS) was derived from mice pretreated with different doses of SC. Cultured human leukemia HL-60 and human gastric adenocarcinoma SGC-7901 cells were used. Inhibition of proliferation was measured using MTT assay. Morphological assessment of apoptosis was performed with fluorescence microscope. DNA fragmentation was assessed by agarose gel electrophoresis and flow cytometry. Expression of bcl-2 protein was measured with immunohistochemistry. Results Exposure of exponentially growing HL-60 cells to mice serum containing 10% SC (pretreated with SC 3, 6, and 12 g/kg) for 48 h resulted in growth inhibition in a dose-dependent manner. Typical morphological changes of apoptosis and DNA fragmentation in HL-60 cells were induced. "Apobodies" in the apoptotic cells were observed, "ladder" pattern of agarose gel electrophoresis of DNA from these cells was revealed, and the percentage of apoptotic cells with fractional DNA content increased from 11.7% to 57.4%. Treatment with SC containing serum decreased the percentage of SGC-7901 cells of bcl-2 protein positive expression from 78.3% to 32.9%. Conclusion SC could induce apoptosis of HL-60 cells and decrease the expression of bcl-2 protein of gastric adenocarcinoma SGC-7901 cells.

HPLC fingerprint of Stellera chamaejasme of Inner Mongolia Region / 中成药

AIM:To establish a HPLC fingerprint of Stellera chamaejasme L. from Inner Mongolia Region. METHODS: The RP-HPLC method was used with Akzonobel Kromasil C_ 18 (250 mm?4.6 mm, 5 ?m) the acetonitrile-0.5% phosphoric acid (gradient elution) was used as mobile phase, analytic time was 60 min, and detective wavelength was at 297 nm, the column temperature of 15℃ were adopted. RESULTS: The HPLC fingerprint of Stellera chamaejasme L. set up showed that 14 peaks were co-possessing in different sources. The results of method validation met technical standard of fingerprint, the similarities of Stellera chamaejasme L. were 0.9 to 1.0. CONCLUSION: The method is stable and reliable with a good reproducibility and provides a reference standard for the quality control of Stellera chamaejasme L. from Inner Mongolia Region.

Quality standard for Euphorbia / 中成药

AIM: Euphorbia is the unprocessed root of Euphorbia ebracteolata Hayata or Euphorbia Fischeriana Steud,or Stellera chamaejasme L.To establish the quality standard for these potent herbs. METHODS: The microscope examination techniques,physio-chemical methods such as HPLC/UV and LC-MS-MS had been used. RESULTS: The content of ebracteolata compund B in E.Fischeriana was 0.01%,and in E.ebracteolata was(0.01%)-0.02%.But the skimmetine,chamaechromone and neochamaejasmin A had not been chekout in both species.The content of skimmetine,chamaechromone and neochamaejasmin A was 0.57%-2.12 %,0.86%-(1.6%),and 0.45%-0.96% in S.chamaejasme.But the ebracteolata compund B had not been chekout in this specie. CONCLUSION: The method can be applied to conventional testing of Euphorbia.

Serum-pharmacological effects of Stellera chamaejasme L. abstract with water on proliferation of gastric adnocacinoma SGC-7901 Cells / 中成药

Objective: To explore the anticancer mechanism of Stellera chamaejasme L. abstracts with water (SCLA). Methods: SCLA drugserum was derived after different time from mice which was orally peradministrated with different dose of SCLA. the effects of the serum on the roliferation of human gastric adnocacinoma SCG7901 cells were observed with MTT assay and clone formation methods.Results: After treatment with the drugserum derived at 1,2,3h from mice pertreated with SCLA(5,10 and 20g?kg -1 ), the MTT formazan and cone formation of gastric adnocacinoma SGC7901 cells were decreased significantly. SCLA from the mice which were given the drugserum 2h later was more effective than others. Conclusions: SCLA had a remarkable inhibitory effects on proliferation of gastric adnocacinoma SGC7901 cells, which may be the main anticancer mechanisms of SCLA.