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Objective: To establish a new method for the quantitative analysis of multi-components by single-marker (QAMS) to simultaneous determine six naphthoquinone components in Arnebia euchroma. Methods: The chromatographic peaks of the main naphthoquinone components in A. euchroma were identified by high resolution LC-MS. The acetyl Shikonin was used as internal marker to calculate the relative correlation factors (RCF) of deoxyshikonin, isobutyrylshikonin, β-acetoxyisovalerate shikonin, and β,β'-dimethylacryloyl shikonin by HPLC, and examine the durability and reproducibility of the RCF. The external standard method and QAMS were compared to determine the six components in A. euchroma. Results: The repeatability of RCF was good. The results calculated with QAMS were consistent with the results by the external standard method. Conclusion: The QAMS method for simultaneously measuring the content of six components is accurate and reliable to evaluate the quality of A. euchroma.
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OBJECTIVE: To establish a method for the simultaneous determination of shikonin, acetylshikonin and β, β-dimethylacrylshikonin in Arnebia euchroma. METHODS: RP-HPLC method was adopted. The determination was performed on Kromasil 100-5 C18 column with mobile phase consisted of acetonitrile-0. 1％ formic acid solution (80: 20, V/V) at the flow rate of 1. 0 mL/min. The detection wavelength was set at 516 nm, column temperature was 25 ℃, and sample size was 10 μL. RESULTS: The linear ranges of shikonin, acetylshikonin and β, β-dimethylacrylshikonin were 0. 404-10. 100 μg/mL(r=0. 999 8), 5. 350-107. 000 μg/mL(r=0. 999 6), 2. 035-40. 700 μg/mL(r=0. 999 8), respectively. The limit of quantitation was 0. 40, 2. 91, 1. 34 μg/mL, and the limit of detection was 0. 12, 0. 87, 0. 40 μg/mL. RSDs of precision, stability and reproducibility tests were all lower than 2. 0％ (n=6). The recovery rate were 99. 12％-104. 18％ (RSD=1. 85％, n=6), 96. 51％-100. 21％ (RSD=1. 43％, n=6), 98. 11％-102. 51％ (RSD=1. 42％, n=6), respectively. CONCLUSIONS: The method is simple, precise, stable and reproducible. It can be used for simultaneous determination of shikonin, acetylshikonin and β, β-dimethylacrylshikonin in A. euchroma.
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Aim Toevaluatethesynergisticeffectof anti-tumor by the pterostilbene and acetylshikonin act-ing on B16F10 cells and investigate the interrelated mechanisms.Methods Theresearchscreenedandan-alyzed the target-related of pterostilbene and ace-tylshikonin by system-pharmacological methods. The proliferative inhibition rate of B16F10 cells were meas-ured by MTT.The apoptosis in B16F10 cells were proved by both cellular morphological and biochemical methods.The expression of apoptotic genes were as-sessed via RT-PCR.The apoptotic rate and cell cycle were measured by flow cytometry.Melanoma models were established in C57BL/6 mice,and the inhibitory rateoftumorgrowthwasmeasured.Results The14 targets of pterostilbene were closely related to cell cy-cle,acetylshikonin′s 12 targets displayed a relationship with apoptosis,and correlated with p53 signaling path-way.Pterostilbene along with acetylshikonin signifi-cantly inhibited cell proliferation of B16F10 cells in a dose-dependent way and resulted a remarkable syner-gistic effect.The apoptotic rate reached highest with a blocked-cell cycle at G1 phase in the co-treatment group.The RT-PCR results showed that the expres-sions of p53,Bax and p21 were up-regulated and the expressions of Bcl-2,CDK2 and Cyclin E were down-regulated with time.The changes of p53,Bax and Bcl-2 were obvious in combined treated group.All treat-ments in vivo showed different tumor inhibition rates while co-treatment group showed highest.Conclusion Pterostilbenecooperatedwithacetylshikonininhibits the proliferation in B16F10 cells,and activates the p53 signaling pathway to induce the B16F10 cells apoptosis and a cell cycle arrest.
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Objective: By examining the secondary metabolites content changes of the low temperature stressed Arnebia euchroma suspension cells to explore the effects of unique environmental factors in genuine regional areas on A. euchroma in non-genuine regional areas. Methods: After stressing A. euchroma suspension cells for 24 h at 4 ℃, rosmarinic acid, lithospermum acid, shikonofuran A, shikonofuran E, acetylshikonin, deoxyshikonin, β,β'-dimethylacryl shikonin, isovalerylshikonin, and total shikonin compounds were measured by HPLC on six time points, one of which was before the stress and counted as 0 h, and others were in 12, 24, 48, 72, and 168 h after the stress. Results: The results showed that the different time points after low temperature stress, similar compounds had the same consistent change trends basically. However, the contents had significant differences. In addition to DAS, the maximum value of all the chemical ingredients were in low stress group. Conclusion: Low temperature stress can promote the accumulation of secondary metabolites in A. euchroma suspension cells, and have an important role in revealing the mechanism of the formation of its genuineness.
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Acetylshikonin, a natural naphthoquinone derivative compound, has been used for treatment of inflammation and cancer. In the present study, we have investigated whether acetylshikonin could regulate the NF-kappaB signaling pathway, thereby leading to suppression of tumorigenesis. We observed that acetylshikonin significantly reduced proliferation of several cancer cell lines, including human pancreatic PANC-1 cancer cells. In addition, acetylshikonin inhibited phorbol 12-myristate 13-acetate (PMA) or tumor necrosis-alpha (TNF-alpha)-induced NF-kappaB reporter activity. Proteome cytokine array and real-time RT-PCR results illustrated that acetylshikonin inhibition of PMA-induced production of cytokines was mediated at the transcriptional level and it was associated with suppression of NF-kappaB activity and matrix metalloprotenases. Finally, we observed that an exposure of acetylshikonin significantly inhibited the anchorage-independent growth of PANC-1 cells. Together, our results indicate that acetylshikonin could serve as a promising therapeutic agent for future treatment of pancreatic cancer.
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Humans , Carcinogenesis , Cell Line , Cell Proliferation , Cytokines , Inflammation , NF-kappa B , Pancreatic Neoplasms , ProteomeABSTRACT
OBJECTIVE: To establish a new chromatography technique to, separate and purify naphthaquinone components from rhizoma of Arnebia euchroma(Royle). METHODS: Medium and low pressure preparative chromatography and Flash column were used to separate and purify the constituents, and their structures were elucidated by physicochemical properties, IR, MS, 1H-NMR and C-NMR. RESULTS: Six monocases were obtained, which were deoxyshikonin, shikonin, isobutylshikonin, acetylshikonin, β-ace-toxyisovalerylshikonin, and β, β-dimethylacrylshikonin. The purities of the 6 compounds analyzed by HPLC were 99.62%, 99.02%, 99.5%, 99.31%, 99.3% and 99.2%, respectively. CONCLUSION: A medium and low pressure preparative chromatography method has been established for the first time which can accurately and rapidly separate monocases from herbs.
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In this report, we describe the cases recovering from oral candidosis by treatment with oral ointment containing antifungal naphthoquinone derivatives. The patients bearing persistent colonization with Candida genera were treated with our domestic oral ointment, three times a day for two weeks or for one month. During the observation period, the oral candidosis was gradually vanished at 3 days after the treatment and disappeared completely at the end of the period. This ointment contains naphthoquinone derivatives, which are constituents of Shikon (root of <i>Lithospermum erythrorhizon</i>), having been investigated against several fungal pathogens. When the biological activity of these compounds was tested against fungi, a wide range of sensitivity was recorded. With the determination of these naphthoquinones by HPLC, this ointment contains about 10 to 100-folds of each effective concentration.<br>This observation demonstrates that the oral ointment containing some antifungal naphthoquinone derivatives would useful for the patients bearing serious oral candidosis.
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300mg%). Conclusion: Acetylshikonin's content was high, radix arnebiae oil colour was very red, It was up to the mustard.