ABSTRACT
This study aimed to evaluate the biological effect and mechanism of Vernonia anthelmintica Injection(VAI) on melanin accumulation. The in vivo depigmentation model was induced by propylthiouracil(PTU) in zebrafish, and the effect of VAI on melanin accumulation was evaluated based on the in vitro B16F10 cell model. The chemical composition of VAI was identified according to the high-performance liquid chromatography quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS). Network pharmaco-logy was applied to predict potential targets and pathways of VAI. A "VAI component-target-pathway" network was established, and the pharmacodynamic molecules were screened out based on the topological characteristics of the network. The binding of active molecules to key targets was verified by molecular docking. The results showed that VAI promoted tyrosinase activity and melanin production in B16F10 cells in a dose-and time-dependent manner and could restore the melanin in the body of the zebrafish model. Fifty-six compounds were identified from VAI, including flavonoids(15/56), terpenoids(10/56), phenolic acids(9/56), fatty acids(9/56), steroids(6/56), and others(7/56). Network pharmacological analysis screened four potential quality markers, including apigenin, chrysoeriol, syringaresinol, and butein, involving 61 targets and 65 pathways, and molecular docking verified their binding to TYR, NFE2L2, CASP3, MAPK1, MAPK8, and MAPK14. It was found that the mRNA expression of MITF, TYR, TYRP1, and DCT in B16F10 cells was promoted. By UPLC-Q-TOF-MS and network pharmacology, this study determined the material basis of VAI against vitiligo, screened apigenin, chrysoeriol, syringaresinol, and butein as the quality markers of VAI, and verified the efficacy and internal mechanism of melanogenesis, providing a basis for quality control and further clinical research.
Subject(s)
Animals , Vernonia/chemistry , Melanins/metabolism , Zebrafish/metabolism , Network Pharmacology , Molecular Docking Simulation , Apigenin/pharmacology , Drugs, Chinese Herbal/pharmacology , Chromatography, High Pressure LiquidABSTRACT
Present study was conducted to standardize callus development and indirect organogenesis from differentexplants of Vernonia anthelmintica (L.) Willd. Among, the various hormonal combinations tested: IndoleAcetic Acid (IAA) and BA and IAA and Kinetin combinations were found to be optimum for inducing callusingwithin a time span of 10 days. Best callus response was observed in 1.5 mg l−1 IAA and 1.5 mg l−1 BA,which produced white friable callus. Best indirect shoot organogenesis was observed in 4 mg l−1 BA and6 mg l−1 kinetin. Elongated shoots when transferred on to half strength Murashige and Skoog (MS) Mediumsupplemented with Indole-3-butyric acid (IBA), rooting was observed. MS medium fortified with 2 mg l−1 IBAshowed better rooting than all other concentrations tested. This concentration produced maximum number ofroots and maximum percentage of rooting. Plantlets developed by indirect organogenesis of V. anthelminticawere successfully acclimatized to field conditions
ABSTRACT
This work aims to establish an HPLC specific chromatogram and determine six components of Vernonia anthelmintica with chlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, scutellarein and luteolin as index components. HPLC analysis was performed on a Waters Xbridge C_(18) column(4.6 mm×250 mm, 5 μm) with gradient elution of acetonitrile-0.05% trifluoroacetic acid solution at a flow rate of 1.0 mL·min~(-1). The detection wave length was 360 nm and the column temperature was 30 ℃. Chemometrics software Chempattern was employed to analyze the data. HPLC specific chromatogram of V. anthelmintica was established and six characteristic peaks were marked. Six characteristic peaks were simultaneously determined by HPLC within 50 min. The contents of the six components in 13 batch samples of V. anthelmintica were 0.14%-0.68%, 0.44%-0.74%, 0.63%-1.01%, 0.14%-0.71%, 0.15%-0.26% and 0.010%-0.030%, respectively. The HPLC specific chromatogram of V. anthelmintica, together with determination of six components showed strong specificity, and it can be used for the quality control of the crude drug.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , Phytochemicals/analysis , Quality Control , Vernonia/chemistryABSTRACT
Objective To establish the chromatographic conditions of isochlorogenic acid A and isochlorogenic acid C in vernonia anthelmintica. Methods By changing the mobile phase,flow rate,column temperature and other chromatographic conditions,the best chromatographic conditions was we pursued to established. Results The linear relationship between the concentration of isochlorogenic acid A and the peak area was between 5. 825-69. 9 μg·mL-1, and the concentration of isochlorogenic acid C,was between 5.15-61.80 μg·mL-1and the peak area was good . The sample recovery rates of the two groups were 98.70%-101.92%(RSD＝1.04%,n＝9)、95.99%-102.52%(RSD＝1.90%,n＝9). Conclusion The method is simple,rapid, accurate and reliable for the determination of isochlorogenic acid A and isochlorogenic acid C in Vernonia anthelmintica and also for the quality control of the raw material.
ABSTRACT
OBJECTIVE: To purify butin and butein from the Vernonia anthelmintica Willd, and preliminary study their effects on proliferation and melanogenesis of A375 human melanoma cells. METHODS: Selica gel and Sephadex LH-20 column chromatography methods were used to separate butin and butein; their effects on proliferation and melanogenesis of A375 human melanoma cells were studied by MTT method, enzyme method and NaOH method. RESULTS: Butin and butein were isolated and identified. In the concentration range of 0.50-10.00 μg · mL-1, butin and butein enhanced the proliferation of A375 human melanoma cells, and the effect of butein was stronger than butin(P < 0.05) ; in the concentration range of 0.10-1.00 μg · mL-1, both butin and butein increased the activity of tyrosinase; in the concentration range of 0.50-5.00 μg · mL-1, butein stimulated melanogenesis, but butin inhibited melanogenesis. CONCLUSION: Butin and butein may be the active components of Vernonia anthelmintica Willd; butein promotes the proliferation and stimulates the melanogenesis of A375 human melanoma cells, however, butin mildly inhibites the melanogenesis.
ABSTRACT
Objective:Vernonia anthelmintica,with complicated components,is a common medicine for the treatment of vitiligo.The purpose of this study was to establish and optimize the separation conditions of high performance liquid chromatography fingerprints(HPLC-FPs) for Vernonia anthelmintica(L.) Willd.Methods: We investigated the HPLC-FPs of different extraction samples by the retention times and relative areas of common peaks in the fingerprints.Results: We established the optimum separation conditions of HPLC-FPs for Vernonia anthelmintica(L.) Willd,and obtained 19 common peaks in the fingerprints.The HPLC-FPs of the 85% ethanol extract were quite similar to those of the semi-bionic extract(SBE) but very different from those of the water extract of Vernonia anthelmintica(L.) Willd.The No.1,4,6,11,12,14 and 16-19 peaks were the endemic ones of the HPLC-FPs of the 85% ethanol extract and SBE of Vernonia anthelmintica(L.) Willd,while No.20-30 were the endemic peaks of the HPLC-FPs of the water extract.Conclusion: The results of our study has provided experimental evidence for further investigation into the constituents absorbed into blood,pharmaceutical technology and pharmacodynamics of Vernonia anthelmintica(L.) Willd.
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OBJECTIVE:To find out a new purifying process of Vernonia Anthelmintica Willd by clarifying agent.METHO-DS:Using orthogonal design L16(45),influcing factors including clarifing agent on extraction process were investigated by detecting the content of total flavone ingredients.RESULTS:Different ratios of concentration volume had significant influence on the content of total flavone ingredients(P
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OBJECTIVE:To investigate the effects of extracts of different constitutes of Vernonia Anthelmintica Willd different on the serum levels of Cu and Zn in mice METHODS:Different groups of mice were given separately with (B) 1/10,(C)1/30,(C′)1/50 doses of LD50 of extracts of different constitutes of Vernonia Anthelmintica Willd for 7 days,and the levels of Cu and Zn in serum were determined RESULTS:The levels of Cu and Zn in serum were different in different groups of mice(P
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Objective To optimize the best condition of semi-bionic extraction for Vernonia anthelmintica.Methods The best condition of the semi-bionic extraction for V.anthelmintica by uniform design was optimized with total HPLC integrate area of common peaks in fingerprints,total flavonoids and dried extract weight as the indexes.Results According to industry production condition,the best technical condition:pH values of 80% ethanol for the 1st,2nd,and 3rd decoctions were in order of 6.0,6.5,and 8.0,and the extraction time were 1,0.5,and 0.5 h,respectively.Conclusion It is better for V.anthelmintica extracted by semi-bionic extraction,and the parametrs are optimized by uniform design.