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OBJECTIVE@#Although Angelica archangelica is a medicinal and aromatic plant with a long history of use for both medicinal and food purposes, there are no studies regarding the antineoplastic activity of its root. This study aimed to evaluate the cytotoxicity and antitumor effects of the crude extract of A. archangelica root (CEAA) on breast cancer.@*METHODS@#The cytotoxicity of CEAA against breast adenocarcinoma cells (4T1 and MCF-7) was evaluated by a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Morphological and biochemical changes were detected by Hoechst 33342/propidium iodide (PI) and annexin V/PI staining. Cytosolic calcium mobilization was evaluated in cells staining with FURA-4NW. Immunoblotting was used to determine the effect of CEAA on anti- and pro-apoptotic proteins (Bcl-2 and Bax, respectively). The 4T1 cell-challenged mice were used for in vivo assay.@*RESULTS@#Using ultra-high-performance liquid chromatography-mass spectrometry analysis, angelicin, a constituent of the roots and leaves of A. archangelica, was found to be the major constituent of the CEAA evaluated in this study (73 µg/mL). The CEAA was cytotoxic for both breast cancer cell lines studied but not for human fibroblasts. Treatment of 4T1 cells with the CEAA increased Bax protein levels accompanied by decreased Bcl-2 expression, in the presence of cleaved caspase-3 and cytosolic calcium mobilization, suggesting mitochondrial involvement in breast cancer cell death induced by the CEAA in this cell line. No changes on the Bcl-2/Bax ratio were observed in CEAA-treated MCF7 cells. Gavage administration of the CEAA (500 mg/kg) to 4T1 cell-challenged mice significantly decreased tumor growth when compared with untreated animals.@*CONCLUSION@#Altogether, our data show the antitumor potential of the CEAA against breast cancer cells in vitro and in vivo. Further research is necessary to better elucidate the pharmacological application of the CEAA in breast cancer therapy.
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Objective: To investigate the effects of psoralen and angelicin on inflammation cytokine expression of human periodontal ligament cells (hPDLCs). Methods: hPDLCs were primarily cultured using tissue explant method. Effects of psoralen and angelicin on the cell viability were tested by CCK-8 assay. hPDLCs were stimulated by Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) after treatment with different concentrations of psoralen and angelicin for 2 h. mRNA expression of IL-1β and IL-8 were determined by real-time PCR. Enzyme-linked immunosorbent assay (ELISA) was used to measure the secretion of IL-1β and IL-8. Results: hPDLCs were cultured successfully by tissue explant method. Psoralen and angelicin (≤ 12.5 μg/mL) did not show significant effects on the cell viability of hPDLCs. Pg-LPS markedly elevated the mRNA expression of IL-1β and IL-8, which could be attenuated by psoralen and angelicin in a dose-dependent manner. Likewise, the up-regulated protein secretion of IL-1β and IL-8 could be significantly blocked by psoralen and angelicin. Conclusion: Psoralen and angelicin could attenuate the inflammatory response of hPDLCs induced by Pg-LPS. Therefore, psoralen and angelicin may act as natural agents to prevent and treat periodontitis.
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Objective·To investigate the effects of psoralen and angelicin on inflammation cytokine expression of human periodontal ligament cells (hPDLCs).Methods· hPDLCs were primarily cultured using tissue explant method.Effects ofpsoralen and angelicin on the cell viability were tested by CCK-8 assay,hPDLCs were stimulated by Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) after treatment with different concentrations of psoralen and angelicin for 2 h.mRNA expression of IL-1β and IL-8 were determined by real-time PCR.Enzyme-linked immunosorbent assay (ELISA) was used to measure the secretion of IL-1β and IL-8.Results · hPDLCs were cultured successfully by tissue explant method.Psoralen and angelicin (≤ 12.5 μg/mL) did not show significant effects on the cell viability of hPDLCs.Pg-LPS markedly elevated the mRNA expression of IL-1β and IL-8,which could be attenuated by psoralen and angelicin in a dose-dependent manner.Likewise,the up-regulated protein secretion of IL-1β and IL-8 could be significantly blocked by psoralen and angelicin.Conclusion · Psoralen and angelicin could attenuate the inflammatory response of hPDLCs induced by Pg-LPS,Therefore,psoralen and angelicin may act as natural agents to prevent and treat periodontitis.
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Objective To study the effect of Angelicin on proliferation activity and anti-aging related protein expression of human HDF cells and its mechanism. Methods According to the random number table method, the cells were divided into blank group, model group, estradiol group, Angelicin group, estrogen receptor antagonist+estradiol group, estrogen receptor antagonist+Angelicin group, and P38 pathway blocker group. Different groups were given the according drugs respectively for 24 h. Except the blank group, all the groups of cells were given UVB irradiation with a dose of 150 mJ/cm2. The MTT assay was used to detect cell proliferation rate. The Western blot was used to detect the expression levels of COLⅠ, MMP-1, ERβ, P38 and p-P38 in cells, and the MMP-1 mRNA expression was detected by real-time fluorescent quantitative PCR.Results Compared with the model group, the proliferation rate of HDF cells significantly increased in Angelicin(10,1,0.1 and 0.01 μmol/L groups)(P<0.01);The protein expression of COLⅠ (0.326 ± 0.006 vs. 0.176 ± 0.007),ERβ(0.281 ± 0.011 vs.0.143 ± 0.006)significantly increased(P<0.01),and the expression of MMP-1(0.256 ± 0.006 vs.0.395 ± 0.006)and p-P38(0.224 ± 0.003 vs.0.318 ± 0.005)significantly decreased (P<0.01) in Angelicin 10 μmol/L group. Compared with 10 μmol/L Angelicin group, the protein expression of estrogen receptor antagonist+Angelicin group ERβ(0.120 ± 0.007 vs.0.281 ± 0.011)significantly decreased and MMP-1mRNA(1.377 ± 0.012 vs.1.024 ± 0.010)significantly increased(P<0.01).Conclusions The Angelicin may degrade MMP-1 through the ER-P38 MAPK signaling pathway,and then promote collagen synthesis, to achieve the purpose of prevention and treatment of photoaging.
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Objective To assay Icariine, Epimedin C, asperosaponin Ⅵ, psoralen and angelicin in Xianlinggubao capsules via multi-wavelength HPLC method.Methods Separation was carried out on Welch Ultimate○R XB-C18 column.The mobile phase was acetonitrile-water system and a linear gradient elution was used.The column temperature was 30 ℃.The detection wavelength for Icariine, Epimedin C,asperosaponin Ⅵ was set at 212 nm, psoralen and angelicin at 246 nm.Results Five components reached baseline separation, the linearity was good when sample size was in the range of 0.008 2-0.328 μg for Icariine(r=0.999 5), 0.055 6-2.224 μg for Epimedin C (r=0.999 6), 0.144 1-5.764 μg for asperosaponin Ⅵ(r=0.999 6), 0.005 4-0.215 2 μg for psoralen(r=0.998 0), 0.006 6-0.265 6 μg for angelicin(r=0.998 5).The average recoveries were 97.59％, 98.58％, 98.11％, 97.86％, 98.22％ respectively.The RSDs of recovery were all less than 2.0％.Conclusion This method is simple, accurate, with good separation, high sensitivity for the assay of multiple components in Xianlinggubao capsule.
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Objective To establish the method for simultaneous determination of epimedium glycoside and other 4 kinds of active ingredients in Xianling Gubao Capsules by three wavelength switching method. Methods An Waters Atlantis T3 C18 column (4.6 mm × 250 mm, 5 μm) was used with the mixture of acetonitrile-0.05％ formic acid solution as the mobile phase in gradient elution (0–5 min, 12％–20％ A; 5–15 min, 20％–55％ A; 15–35 min, 55％ A;35–55 min, 55％–76％ A; 55.1 min, 12％ A; 55.1–60 min, 12％ A). Detection wavelength was as follow: 0–30 min, 212 nm; 30–42 min, 246 nm; 42–60 min, 270 nm. The flow rate was 1.0 mL/min. The column temperature was 30 ℃. Results The calibration curves of asperosaponin Ⅵ, psoralen, angelicin, epimedin C, and icariin were in good linearity among the ranges of 101.6–3048 ng, 8.7–261 ng, 7.9–237 ng, 117.2–3516 ng, 78–2340 ng, respectively. In the instrument precision test, stability test, repeatability test, the RSD was less than 3％. The average recoveries were 99.83％, 100.35％, 100.59％, 100.60％, 99.72％, respectively, and all the RSD were less than 3％. Conclusion The method is sensitive, accurate, and separation effect is good, which can provide a basis for quality evaluation standard of Xianling Gubao Capsules.
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Objective: To determine asperosaponin VI, psoralen, and angelicin in Xianling Gubao Capsules (XGC) via multi- wavelength HPLC method. Methods: Separation was carried out on Welch Ultimate® XB-C18 column. The mobile phase was acetonitrile-water system and a linear gradient elution was used. The column temperature was 30℃. The detection wavelength for asperosaponin VI was set at 212 nm, those for psoralen and angelicin were set at 246 nm. Results: Three components reached baseline separation, the linearity was good when sample volumes were in the ranges of 144.1-5 764.0 for asperosaponin VI (r = 0.999 6), 5.4-215.2 (r = 0.998 0) for psoralen, and 6.6-265.6 ng (r = 0.998 5) for angelicin. The average recoveries of asperosaponin VI, psoralen, and psoralen were 98.11%, 97.86%, and 98.22%, respectively. The RSDs of recoveries were all less than 2.0%. Conclusion: The method is simple and accurate and has good separation, with high sensitivity and good efficiency for the determination of more-index components in XGC.
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Objective: To investigate the positioning based on the relative retention time of fingerprinting and to establish a new quality evaluation method for traditional Chinese medicine preparations, using one chemical reference substance to calcutate multi- components simultaneously. Methods: Employed icariin as the maker component, icariin relative correction factors (RCF) of epimedin C to icariin, asperosaponin VI to icariin, psoralen to icariin and angelicin to icariin were ealeatated in the chromatographic conditions for determination of the four components in Xianling Gubao Capsule (XGC). The contents of icariin were determined by external standard method, and those of epimedin C, asperosaponin VI, psoralen and angelicin were calculated by icariin and their RCF. The accuracy of the new method was evaluated by comparing the relative retention times and calculating the content which using different brands columns for determination. Results: The analysis methods were established,the linearity was good when sample volume was in the range of at 8.2—328.0 ng for icariine(r = 0.999 5), 0.055 6—2.224 μg for epimedin C (r = 0.999 6), 0.144 1—5.764 μg for asperosaponin VI (r = 0.999 6), 5.4—215.2 ng (r = 0.998 0) for psoralen, 6.6—265.6 ng (r = 0.998 5) for angelicin. The average recoveries of asperosaponin VI, psoralen and psoralen were 97.59%, 98.58%, 98.11%, 97.86%, 98.22%, respectively. The RSDs of recovery were all less than 2.0%; There has been no significant difference between the calculated contents and the determined contents, according to the angle cosine value. Conclusion: The method can control the components without providing epimedin C, asperosaponin VI, psoralen, and angelicin reference. The method is not only save reference substance and medicine resources, but also suitable quality evaluation pattern for TCM preparation. This new method made fingerprinting more meaningful in TCM quality control.
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Objective To develop a HPLC method for psoralenand angelicin content assay in Zhuanggu granules .Meth-ods Agilent Zorbax C18 column(4 .6 mm × 250 mm ,5 μm) was used with methanol and water (45∶55) as mobile phase .The detection wavelength was set at 245 nm .The flow rate was 1 .0 ml/min and column temperature at 25 ℃ .The injection volume was 10 μl .Psoralenand angelicin were extracted with 70% ethanol under water bath .Results The linearity was obtained over 3 .75~40 μg/mlfor psoralenand angelicin .The RSDs were less than 2% .The average recovery was between 94% and 105% . Conclusion This simple and accurate HPLC method was suitable for the quality control of Zhuanggu granules .
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OBJECTIVE:To establish a method for simultaneous determination of 4 psoralen compounds in Buwu tincture. METHODS:HPLC was performed on the column of Dikma Diamonsil C18 with mobile phase of acetonitrile-0.2% Acetic acid by gradient elution at flow rate of 1 ml/min,detection wavelength was 246 nm,column temperature was 30 ℃,and the injection vol-ume was 15 μl. RESULTS:The linear range was 13.00-208 μg/ml for angelicin,26.00-416 μg/ml for bavachin,24.50-392 μg/ml for psoralidin and 37.88-606 μg/ml for isobavachalcone,respectively(r≥0.999 6);RSDs of precision,stability and reproducibility tests were less than 2.00%;recoveries were 95.22%-97.23%(RSD=0.87%,n=6),100.24%-104.64%(RSD=1.62%,n=6), 102.28%-104.39%(RSD=1.47%,n=6)and 97.68%-100.17%(RSD=0.97%,n=6),respectively. CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Buwu tincture.
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Objective To establish the quality standard of Huangqi-Changyu'an capsule. Method Thin layer chromatography was used to identify psoralen, evodia mtaecarpa, and nutmeg in Huangqi-changyu'an capsule. HPLC was used to determine the content of psoralen. A Hypeisil BDS C18 column(200 mm×4.6 mm,5 μm) was used with mobile phase consisted of acetonitrile-water (30:70) at the rate of 1 ml/min. The detection wavelength was set at 245nm with the column at room temperature. Results The liner range of psoralen was 3.572~53.58 μg/ml (r=0.999 9) with average recoveries of 99.04%, and the RSD was 0.8%. The liner range of isopsoralen was 3.796~56.94 μg/ml (r=0.999 9) with average recoveries of 99.20%, and the RSD was 0.6%. Conclusion This method is sentivive, accurate and rapid for the quality control of Huangqi-Changyu'an capsule.
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BACKGROUND: Monofunctional psoralens plus UVA radiation are not severely phototoxic and have less mutagenic activity than bifunctional psoralens plus UVA radiation. OBJECTIVE: The purpose of this study was to evaluate pigment producing effect using various concentrations(0.02%, 0.1%, 0.5%) of monofunctional psoralens such as angelicin, khellin and comparing it's effect with TMP in topical photochemotherapy. METHOD: Ninty three C57BL mice were painted with either angelicin, khellin or TMP solution in concentrations of 0.02%, 0.1% and 0.5% each and were UVA irradiated. Skin biopsies were performed at 1,3,5 weeks after UVA irradiation. The pigment producing effects were measured by the number, area and perimeter of the melanocytes after topical PUVA. RESULTS: The comparison of melanocyte numbers between different psoralens after five weeks of photochemotherapy showed a significant difference in decreasing order of TMP, khellin and angelicin. The area and perimeter of melanocytes were larger in the TMP group after five weeks photochemotherapy than the other group. However in the khellin and angelicin group, the area and perimeter of melanocytes were not increased by increasing the frequency of the UVA irradiation. CONCLUSION: The number, area and perimeter of melanocytes after topical PUVA increased in the TMP group compared to angelicin or khellin group. We expect the clinical application of angelicin and khellin in vitiligo is possible considering the result of the study of pigment producing effect with a higher concentration and higher dose of UVA.