ABSTRACT
In this study, the toxicological/pharmacological research method of "quantity-weight-evidence" network was first proposed and practiced to supplement the existing methodology of network toxicology. We transformed the traditional qualitative network into a quantitative network in this study by attributing weights to toxic component content and target frequency, which improved the reliability of data and provided a research idea for the systematic safety evaluation and toxicological research of Chinese medicinal herbs. Firstly, 50% ethanol extract of Dysosma versipellis(DV) was administrated to rats via gavage and the potential hepatotoxic components were identified by serum pharmacochemistry. Then, the component targets were obtained from SwissTargetPrediction, PharmMapper and other online databases, and the target weights were given according to the relative content of components and target fishing frequency. Meanwhile, the targets of hepatotoxicity were predicted from online databases such as Comparative Toxicology Database(CTD) and GeneCards. Subsequently, protein-protein interaction analysis and KEGG pathway enrichment were performed with the STRING database. Finally, the quantitative network of "toxic components-weighted targets-pathways" was constructed. Eleven potential toxic compounds were predicted, including podophyllotoxin, podophyllotoxone, deoxypodophyllotoxin, and 6-methoxypodophyllotoxin. A total of 106 hepatotoxic targets and 65 weighted targets(e.g., Cdk2, Egfr, and Cyp2 c9) were identified. The results of Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment showed that these targets could act on PI3 K-AKT, MAPK, and Ras signaling pathways to play a role in inflammatory response and oxidative stress. However, traditional network toxicology showed that 51 targets such as AKT1, Alb, and Stat3 may lead to hepatotoxicity by mediating inflammation and cell proliferation. In conclusion, we proposed "quantity-weight-evidence" network toxicology in this study and used it to study the mechanism of DV-induced hepatotoxicity in rats. This study confirms the feasibility of this new methodology in toxicological evaluation and further improves the systematic evaluation of the safety of Chinese medicinal herbs.
Subject(s)
Animals , Rats , Chemical and Drug Induced Liver Injury/etiology , Drugs, Chinese Herbal/toxicity , Ethanol , Medicine, Chinese Traditional , Molecular Docking Simulation , Reproducibility of ResultsABSTRACT
To determine the inhibitory effect of endophytic fungi from Dysosma versipellis on HIV-1 IN-LEDGF/p75 interaction,the protein-protein interaction between human immunodeficiency virus type 1( HIV-1) integrase and lens epithelial growth factor p75 protein( LEDGF/p75) was used as a target. The homogeneous time-resolved fluorescence( HTRF) technique was used in the inhibitory activity assay. The results showed that eight endophytic fungi with anti-IN-LEDGF/p75 interaction activity were screened out from fifty-three strains with different morphological characteristic. Among them,106 strain showed strong inhibitory activity against HIV-1 IN-LEDGF/p75 interaction with IC50 value of 5. 23 mg·L-1,and was identified as a potential novel species of Magnaporthaceae family by the analyses of ITS-rDNA,LSU and RPB2 sequences data. This study demonstrated that potential natural active ingredients against the HIV-1 IN-LEDGF/p75 interaction exist in the endophytic fungi of D. versipellis. These results may provide available candidate strain resources for the research and development of new anti-acquired immunodeficiency syndrome drugs.
Subject(s)
Humans , Berberidaceae , Microbiology , Endophytes , Fungi , Chemistry , HIV Integrase , Metabolism , HIV-1 , Intercellular Signaling Peptides and Proteins , Metabolism , Protein BindingABSTRACT
Using the White as basic medium, the effects of the exogenous IBA and endophytic fungal elicitor on the growth of in vitro roots cultures of Dysosma versipellis and production of podophyllotoxin were investigated in this study. The results showed that the IBA and the endophytic fungus Zasmidium syzygii elicitor could increase the content of podophyllotoxin of in vitro roots of D. versipellis after 3 weeks. The White medium added with 3 mg·L~(-1) IBA induced the highest increase of podophyllotoxin(1 830.86 μg·g~(-1)), which was 2.07 folds greater than the control, and followed by 1.5 mg·L~(-1) IBA, fungal elicitor, 1 mg·L~(-1) IBA, 0.5 mg·L~(-1) IBA and 4.5 mg·L~(-1) IBA, which was 1.82, 1.71, 1.63, 1.43 and 1.1 folds greater than the control, respectively. The results also showed that the growth of roots was certain positively correlated with the change of IBA concentration. Therefore, 3 mg·L~(-1) IBA was the most suitable for the production of podophyllotoxin in the in vitro roots of D. versipellis, and the stimulating effect of Z. syzygii fungal elicitor was between 1.5 mg·L~(-1) and 1 mg·L~(-1) IBA, which was a potential natural elicitor to induce the accumulation of podophyllotoxin in future production.
Subject(s)
Ascomycota , Berberidaceae , Chemistry , Endophytes , Plant Roots , Podophyllotoxin , Tissue Culture TechniquesABSTRACT
Objective To explore a new identification method for medicinal materials of Dysosma, and analyze the second internal transcribed spacer (ITS2) barcode sequences of Diphylleia sinensis and Dysosma versipellis, Sinopodophyllum hexandrum, Dysosma difformis and Dysosma pleiantha in five kinds of podophyllum. Methods The ITS2 of ribosomal DNA of medicinal materials of podophyllum was amplified and sequenced by bi-directional sequencing of PCR products. Sequence assembly and consensus sequence generation were performed by using CodonCode Aligner. Phylogenetic study was performed using software MEGA 5.1 in accordance with Kimura-2-parameter (K2P) model. Genetic distances were calculated and analyzed and the phylogenetic tree was constructed by using the neighbor-joining (NJ) method. Results There were significant differences among five kinds of Dysosma. Their maximum intraspecific genetic distance (K2P distance) was far lower than their minimum interspecific genetic distance with the other species. In the cluster dendrogram, all species showed monophyletic. Conclusion ITS2 sequence as DNA barcoding technique can be used to identify Chinese herbal materials of Dysosma.
ABSTRACT
This study was conducted to investigate the chemical constituents in the root of Dysosma versipellis (Hance) M. Cheng. The constituents were isolated by silica gel, lichroprep RP-C18 and pharmadex LH-20 column chromatography and the IR, MS, NMR, 2D-NMR spectroscopic analysis were employed for the structural elucidation. Ten compounds were isolated from the 95% ethanol extract of Dysosma versipellis, their structures were elucidated as dysoverine D (1), dysoverine F (2), dysoverine A (3), podoverine A (4), α-peltatin (5), rutin (6), kaempferol-3-O-β-D-glucopyranoside (7), quercetin-3-O-β-D-glucopyranoside (8), kaempferol (9) and quercetin (10). Compound 2 is a new compound, and compounds 1 and 3-6 were isolated from this plant fo r the first time.
ABSTRACT
Various chromatographic techniques, including silica gel column chromatography, Sephadex LH-20, preparative thin-layer chromatography, and preparative HPLC, were employed to isolate the chemical constituents from callus cultures of Dysosma versipellis. Structures of the compounds were elucidated based on UV, IR, MS and NMR spectroscopic data analysis. Totally, seven flavonoid glycosides were isolated from the 95% ethanol extract of the callus cultures and identified as kaempferol-3-O-[6″-(3″'-methoxy)-malonyl]-β-D-glucopyranoside(1), kaempferol-3-O-(6″-O-acetyl)-β-D-glucopyranoside(2), kaempferide-3-O-β-D-glucopyranoside(3), kaempferol-3-O-β-D-glucopyranoside(4), isoquercitrin(5), quercetin-4'-O-β-D-glucopyranoside(6) and kaempferol-3-(6″-malonyl)-β-D-glucopyranoside(7), respectively.All these compounds were isolated from callus cultures of D. versipellis for the first time.Compounds 1, 2, 3, 6 and 7 were firstly obtained from plant materials of D. versipellis, and compound 1 was a new compound.
ABSTRACT
Secoisolariciresinol dehydrogenase (SDH) is a key enzyme involved in the biosynthetic pathway of podophyllotoxin.In this study, two SDH candidate genes,SO282 and SO1223, were cloned from callus of Dysosma versipellis by homology-based PCR and rapid amplification of cDNA end (RACE).The SDH candidate genes were expressed in Escherichia coli and the subsequent enzyme assay in vitro showed that recombinant SO282 had the SDH activity. These results pave the way to the follow-up investigation of the biosynthetic of podophyllotoxin.
ABSTRACT
OBJECTIVE: To study the lignans constituents from callus culture of Dysosma versipellis (Hance) M. cheng ex Ying.
ABSTRACT
OBJECTIVE: To examine the diversity and antimicrobial activities of the endophytic fungi of Dysosma versipellis (Hance) M. Cheng. METHODS: The identification of the fungi was carried out by rDNA-ITS sequence analysis. The antimicrobial activities of the isolated endophytic fungi were determined by agar block test. RESULTS: Nineteen strains of various endophytes were obtained from the roots, stems and leaves of D. versipellis, which were molecularly identified to belong to eight catalogues and thirteen genuses. The DV04 had remarkable antimicrobial activities to Staphylococcus aureus, Escherichia coli, and Candida albicans. Morphological and molecular identification demonstrated that DV04 represented Leptodontidium sp.. CONCLUSION: The endophytic fungi in Dysosma versipellis are diverse and rich. Some strains such as DV04 deserve further research as an alternative antimicrobial resource.
ABSTRACT
Objective To observe the pathological changes of major organs in rats with acute Dysosma versipellis poisoning and investigate the toxic mechanismand the injuries of target tissues and organs. Methods Forty Sprague-Daw ley (SD) rats were random ly divided into three experimental groups, which were given the gavage with 0.5, 1.0 and 2.0LD50 doses of Dysosma versipellis decoction, and one con-trol group, which was given the gavage with 1.0LD50 dose of normal saline. The rats were sacrificed 14 days after Dysosma versipellis poisoning and sam ples including brain, heart, liver, lung, and kidney were taken. After pathological process, the pathological changes of the major organs and tissues were observed by light microscope and electron microscope. The experimental data were statistical analyzed by x2 test. Results The observations of light microscopy: loose cytoplasmof neurons with loss of most Nissl bodies;swelling of m yocardial cells with disappearance of intercalated disk and striations;hepato-cellular edema with ballooning degeneration; and swelling epithelial cells of renal proximal convoluted tubule with red light coloring protein-like substances in the tube. The observations of electron microscopy:the structures of cell mem brane and nuclear mem brane of neurons were destroyed;cytoplasmof neurons, obvious edema;and most organelles, destroyed and disappeared. The mortalities of rats after acute poi-soning of the four groups increased with doses (P<0.05). Conclusion Acute Dysosma versipellis poisoning can cause multi-organ pathological changes. There is apositive correlation between the toxic effect and the dosage. The target tissues and organs are brain (neurons), heart, liver and kidney.
ABSTRACT
Objective: To study the chemical constitutents from the roots and stems of Dysosma versipellis. Methods: Silica gel column, Sephadex LH-20, and reverse phase HPLC chromatography were used to isolate the chemical constituents, and the structures of the isolated compounds were elucidated by spectral analysis and physicochemical properties. Results: Fourteen compounds were isolated from EtOAc and n-butanol fraction of the 95% ethanol extract in the roots and stems of D. versipellis, including 10 lignans, i.e. 4′,5′- didemethylpodophyllotoxin (1), podophyllotoxin (2), 4′- demethylpodophyllotoxin (3), diphyllin (4), podophyllo-toxone (5), picropodophyll-otoxin-4-O-β-D-glucoside (6), 4′- demethylpodophyllotoxin-4-O-glucoside (7), diphyllin-4-O-β-D-glucoside (8), podophyllotoxin-4-O-β-D-glucoside (9), and dysosmarol (10); three flavonoids, i.e. kaempferol (11), quercetin (12), and kaempferol-3-O-β-D- glucoside (13); and one sterol, i.e. β-sitosterol (14). Conclusion: Compound 1 is a new natural product, and compounds 7 and 8 are isolated from the plants of Dysosma Woodson for the first time.
ABSTRACT
AIM: To establish the quality standard of Guijiu(Rhizome or root of Podophyllum emodi(Wall.) Ying,Dysosma versipellis(Hance) M.Cheng or Dysosma pleiantha(Hance) Woodson.which herbs contained lignans including podophyllotoxin. METHODS: The microscopic examination, physio-chemical method such as LC-MS had been used. RESULTS: These methods can be used for the identification and quantitative analysis of Guijiu. CONCLUSION: The method proposes microscopic examination technique and physio-chemical method such as LC-MS fingerprint and LC-MS-MS for the identification and quantitative analysis of the herbs.