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Aim To investigate the molecular mechanisms of alcohol extracts of Euphorbia fischeriana steud. against hepatocellular carcinoma (HCC) through a combination of network pharmacology analysis and experimental validation. Methods The active ingredients and targets of alcohol extracts of Euphorbia fischeriana steud. were determined through TCMSP, Swiss ADME, Swiss Target Prediction database and references. The databases DisGeNET and GeneCards were employed to screen potential HCC-related genes. Venny platform, STRING platform and Cytoscape software were applied to construct active ingredient-target-disease and protein-protein interaction (PPI) network maps. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses were performed using the DAVID database. To assess the effects of Euphorbia fischeriana steud. alcohol extracts on BEL-7402 cells, the proliferation and apoptosis were detected by CCK-8, EdU and flow cytometry assays, and the related protein levels of JAK2/STAT3 pathway were analyzed by Western blot. Additionally, H22 hepatocellular carcinoma mouse model was used to evaluate the in vivo efficacy of Euphorbia fischeriana steud. alcohol extracts. Results A total of 916 HCC targeted genes, 30 active ingredients containing the related 567 potential targeted genes, and 115 intersection targets of disease and compounds were obtained. KEGG enrichment analysis identified JAK2/STAT3 signaling as a critical pathway. In vitro experiments showed the alcohol extracts of Euphorbia fischeriana steud. could inhibit proliferation, promote apoptosis and suppress JAK2/STAT3 signaling pathway in a dose-dependent manner in BEL-7402 cells. In addition, the alcohol extracts of Euphorbia fischeriana steud., either alone or in combination with sorafenib, dramatically blocked tumor growth in in vivo tests. Conclusions Euphorbia fischeriana steud. alcohol extracts have anti-cancer effects in HCC, and the molecular mechanisms may be connected to the regulation of JAK2/STAT3 signaling pathway.
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BACKGROUND: Euphorbia fischeriana Steud is a very important medicinal herb and has significant medical value for healing cancer, edema and tuberculosis in China. The lack of molecular markers for Euphorbia fischeriana Steud is a dominant barrier to genetic research. For the purpose of developing many simple sequence repeat (SSR) molecular markers, we completed transcriptome analysis with the Illumina HiSeq 2000 platform. RESULTS: Approximately 9.1 million clean reads were acquired and then assembled into approximately 186.3 thousand nonredundant unigenes, 53,146 of which were SSR-containing unigenes. A total of 76,193 SSR loci were identified. Of these SSR loci, 28,491 were detected at the terminal position of ESTs, which made it difficult to design SSR primers for these SSR-containing sequences, and the residual SSRs were thus used to design primer pairs. Analyzing the results of these markers revealed that the mononucleotide motif A/T (44,067, 57.83% of all SSRs) was the most abundant, followed by the dinucleotide type AG/CT (9430, 12.38%). Using 100 randomly selected primer pairs, 77 primers were successfully amplified in Euphorbia fischeriana Steud, and 79 were successfully amplified in three other related species. The markers developed displayed relatively high quality and cross-species transferability. CONCLUSIONS: The large number of EST-SSRs exploited successfully in Euphorbia fischeriana Steud for the first time could provide genetic information for research on linkage maps, variety identification, genetic diversity analysis, and molecular marker-assisted breeding.
Subject(s)
Euphorbia/genetics , High-Throughput Nucleotide Sequencing/methods , Plants, Medicinal , Genetic Variation , Genetic MarkersABSTRACT
Euphorbiae Ebracteolatae Radix is the dry root of plant Euphorbia fischeriana or E. ebracteolata of Euphorbiaceae, which is a widely utilized natural medicine with broad development prospect. It has been discovered that Euphorbiae Ebracteolatae Radix contains several biologically active constituents, among which diterpenoids are the most important. The diterpenoids of Euphorbiae Ebracteolatae Radix contain eight types including abietane-type, tigliane-type, pimarane-type, rosane-type, cembrane-type, ent-kaurane-type, ingenane-type and atisane-type. Diterpenoids of Euphorbiae Ebracteolatae Radix have significant anti-tumor, anti-inflammatory, anti-viral and other pharmacological activities. In this paper, the chemical constituents of diterpenoids and pharmacological activities of Euphorbiae Ebracteolatae Radix in recent years were reviewed in order to provide references for better developing the resources of Euphorbiae Ebracteolatae Radix and its clinical application.
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Objective To establish a method for determination of ebracteolatain A content in the root of traditional Chinese medicine white Langdu, including Euphorbia fischeriana Steud and Euphorbia ebracteolata Hayata. Methods An HPLC-DAD method was created for determination at the following condition:the column was Agilent Zorbax SB-C18 (4.6 mm×150 mm, 5 μm);mobile phase was acetonitrile:0.1% formic acid in water=55:45(V:V), isocratic elution, the flow rate was 1.0 mL·min-1, the temperature of column was 25°C, the detection wavelength was set at 290 nm, the injection volume was 20 μL, and the running time was 20 min. Results Ebracteolatain A was separated from the interference in the baseline, and the linear range was 4.545-227.3 μg·mL-1, with the linear correlation being 0.9999 for ebracteolatain A. The result of intra-day and inter-day precisions were both within 2%(n=3), and the average recovery was (99.14±3.4)%(n=6). The content of ebracteolatain A in two batches of Euphorbia fischeriana Steud and Euphorbia ebracteolata Hayata were (56.73±1.09) μg/g, (18.98±2.11) μg/g and (235.2±2.4) μg/g (n=6), respectively. Conclusion The present method is simple, rapid, accurate and convenient for determination of ebracteolatain A in the root of traditional Chinese medicine white Langdu.
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Objective: To study the chemical constituents from the aerial part of fresh Euphorbia fischeriana. Methods: Fourteen compounds were isolated and purified by silica gel column chromatography, HPLC and recrystallization methods, and their structures were identified by spectral analysis. Results: Fourteen compounds were isolated and identified as 23 (E)-25-methoxycycloart-23-en-3β-ol (1), 24-hydroperoxycycloart-25-en-3β-ol (2), 25-hydro-peroxycycloart-23-en-3β-ol (3), lupeol (4), 24-methylenecycoartanol (5), euphol (6), 24-methylenecycloartane-3, 28-diol (7), obtusifoliol (8), phytol (9), jolkinolide A (10), 2, 4-dihydroxy-6-methoxy-3-methyl-1-acetophenone (11), 3-oxo-24-methylenecycloarane (12), 12-deoxyphorbol-13-hexadecanoate (13), and butyrospermol (14). Conclusion: Compound 9 is obtained from the plants in Euphorbia L. for the first time, and compounds 1-3 and 7-8 are isolated from E. fischeriana for the first time.
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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.