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OBJECTIVE： To study the effects of ethanol extract of Sanguis Draconis on the survival of perforating flap model in rats and PI3K/Akt/eNOS pathway. METHODS： Perforating flap model was established by cutting off surrounding vessels and keeping one perforator. After modeling， the rats were divided into model group （external use， normal saline） and ethanol extract of Sanguis Draconis （EESD， the content of dracorhodin was 75.08 mg/g） group （external use， 0.21 g/cm2）， with 10 rats in each group. They were given relevant medicine for consecutive 7 days， once a day. The flap survival rate and flap microvessel density were determined after given relevant medicine 7 days. Human umbilical vein endothelial cells （HUVECs） were reoxygenated and glycoconjugated 16 h after hypoxia and hypoglycemia to establish oxygen-glucose deprivation/oxygen-glucose recovery model of HUVECs. After modeling， model cells were divided into normal group， model group， dracorhodin high-concentration， medium- concentration and high-concentration groups （2.5， 1.0， 0.5 μg/mL）. After reoxygenated and glycoconjugated for 24 h， cells morphology was observed by microscope； cell viability and the content of NO were detected by MTT assay and colorimetry. mRNA expression of Akt， PI3K and eNOS， PI3K protein expression， the phosphorylation of Akt and eNOS protein were determined by RT-PCR and Western blot assay. RESULTS： In rat experiment， compared with model group， flap survival rate and microvessel density of rats were increased significantly in EESD group （P＜0.01）. In cell experiment， compared with normal group， the survival rate of HUVEC， NO content， mRNA expression of PI3K， Akt， eNOS，PI3K protein expression， the phosphorylation of Akt and eNOS protein were decreased significantly （P＜0.05 or P＜0.01）. Compared with model group， dracorhodin high-concentration， medium-concentration and high-concentration groups survival rate of HUVEC cells， NO content， mRNA expression of PI3K， Akt and eNOS， PI3K protein expression， the phosphorylation of Akt and eNOS protein were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： The survival rate of perforating flap model in rat can be increased by treating with EESD， the mechanism of which may be associated with the activation of PI3K/Akt/eNOS pathway to protect endothelial cells.
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Objective To investigate the relationship between powder particle size and dissolution of Xuejie Sanqi Jiegu Paste (XSJP) in vitro. Methods The particle size of three kinds of XSJP was measured by laser particle size instrument. The hydroxysafflor yellow A, asperosaponin VI, ginsenoside Rg1, ginsenoside Rb1, notoginsenoside R1, dracorhodin were taken as indexes, and the feature of dissolution for the above three kinds of XSJP was examined by grouting method. The correlation between the powders in different particle sizes and their corresponding dissolution was analyzed by software SPSS 17.0. Results The particle size of the three kinds of powders was correlated with the dissolution of index components. The powders in different particle sizes were negatively correlated with the relative cumulative dissolution of hydroxysafflor yellow A, asperosaponin VI, ginsenoside Rg1, ginsenoside Rb1 and notoginsenoside R1. The cumulative dissolution of superfine powder was respectively increased by 6.8%, 16.6%, 6.6%, 6.7%, and 5.4%, while the dracorhodin had no direct correlation. Conclusion There is a correlation between the particle size and dissolution of Chinese medicine with fibre which contains cellular structure.
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Objective:To determine the content of dracorhodin and curcumin in Dieda pills by UPLC .Methods:A UPLC method was adopted.The determination was performed on a Waters Acquity UPLC BEH C 18 column(100 mm ×2.1 mm,1.7 μm) with the mobile phase consisting of acetonitrile-0.05 mol· L-1 NaH2PO4(50 ∶50).The detection wavelength was 440 nm for dracorhodin and 431 nm for curcumin, the column temperature was 30℃and the flow rate was 0.1 ml· min-1 .Results:There was a good linear range of 0.001 8-0.036 4 μg(r=0.999 9)for dracorhodin and 0.000 8-0.015 6 μg(r=0.999 9) for curcumin.The average recovery for dracorhodin was 97.94%(RSD=0.89%) and that for curcumin was 98.45%(RSD=0.91%).Conclusion:The method is simple, rapid and reproducible ,which can be used for the determination of dracorhodin and curcumin in Dieda pills .
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OBJECTIVE: To establish methods for the test of chemical dyes and quantitative determination of dracorhodin perchlorate in Guanmaining tablets. METHODS: HPLC-UV-MS/MS was used for the test of chemical dyes in Guanmaining tablets, and the concentration of dracorhodin perchlorate was determined by HPLC. In the test of chemical dyes, Agilent Zorbax-SB C18 (4.6 mm × 250 mm, 5 μm) was used as stationary phase, the mixture of 0.02 mol · L-1 ammonium acetate and acetonitrile was used as mobile phase in gradient elution mode, and the UV-Vis detection wavelength was set at 520 nm. ESI+ was used as ion source, and the MS/MS spectra of the analytes were recorded to confirm the positive results. In the quantitative analysis of dracorhodin perchlorate, Phenomenex Luna C18(4.6 mm × 250 mm, 5 μm) was used as stationary phase, the mixture of acetonitrile and 0.05 mol · L-1 sodium dihydrogen phosphate solution (37:63) was used as mobile phase, and the detection wavelength was set at 440 nm. RESULTS: Among 56 batches of Guanmaining tablets, 7 batches showed positive results of sudan I, sudan IV, and/or 808 scarlet. A good linear relationship was obtained in the range of 0.408 0 - 816.6 μg · mL-1 for dracorhodin perchlorate, with the linear regression equation of y = 1.746 1 × 107ρ +2 419.9(r = 1.000 0); the average recovery was 98.37% (RSD = 1.2%, n =6). The content of dracorhodin perchlorate in Guanmaining tablets ranged from 0.01 to 0.25 mg · tablet-1. CONCLUSION: The method is simple, accurate and rapid, and can be used for the quality control of Guanmaining tablets.
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Objective:To establish a method to determine dracorhodin in Gushang Dieda Zhitong capsules by HPLC. Methods:An HPLC method was used for the determination of dracorhodin, a Shimadzu VP-ODS C18 column was used,and acetonitrile-0. 05 mol ·L-1 sodium dihydrogen phosphate solution(35∶65)was the mobile phase. The flow rate was 1.0 ml·min-1. The detection wave-length was at 440nm. Results:The linear range of dracorhodin was 0. 047-0. 234 μg(r=0. 999 9). The average recovery was 99. 9%with RSD of 1. 5%. Conclusion:The method is simple, rapid, accurate and specific. It can be used in the quality control of Gushang Dieda Zhitong capsules.
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The growth inhibition and pro-apoptosis effects of dracorhodin perchlorate on human prostate cancer PC-3 cell line were examined.After administration of 10-80 μmol/L dracorhodin perchlorate for 12-48 h,cell viability of PC-3 cells was measured by MTT colorimetry.Cell proliferation ability was detected by colony formation assay.Cellular apoptosis was inspected by acridine orange-ethidium bromide fluorescent staining,Hoechst 33258 fluorescent staining,and flow cytometry (FCM) with annexin V-FITC/propidium iodide dual staining.The results showed that dracorhodin perchlorate inhibited the growth of PC-3 in a dose- and time-dependent manner.IC50 of dracorhodin perchlorate on PC-3 cells at 24 h was 40.18 μmol/L.Cell clone formation rate was decreased by 86％ after treatment with 20 μmol/L of dracorhodin perchlorate.Some cells presented the characteristic apoptotic changes.The cellular apoptotic rates induced by 10-40 μmol/L dracorhodin perchlorate for 24 h were 8.43％ to 47.71％ respectively.It was concluded that dracorhodin perchlorate significantly inhibited the growth of PC-3 cells by suppressing proliferation and inducing apoptosis of the cells.
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Objective To control the quality of Shujin Jiegu tablets by determining the content of dracorhodin in the preparation by HPLC.Method With dracorhodin as index,the content of dracorhodin in the preparation was determined by HPLC.Results Dracorhodin showed a good linear relationship in the range of 1.53～7.66 ?g,r =0.999 9.The average recovery was 98.64%,RSD=1.12%.Conclusion This method is simple,rapid,repeatable,and applicable to the quality control of the preparation.
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OBJECTIVE:To prepare powder of promoting blood circulation to arrest pain and to establish its quality con?trol method.METHODS:Qualitative identification of panax pseudo-ginseng,frankincense and dragon blood in the powder of promoting blood circulation to arrest pain was performed by TLC method.The content of dracorhodin in the dragon blood was determined by TLC Scanning Method.RESULTS:Good linear relationship was achieved when the sample size of dracorhodin perchlorate remained within the scope of0.6?g～3.2?g(r=0.9999).The average recovery rate was98.22%(RSD=1.53%).CONCLUSION:This preparation method is feasible,and the quality control method is simple,accurate and reproducible.
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OBJECTIVE: To determine the content of dracorhodin in Diedaqili Tablets by HPLC. METHODS: The separation was performed on Diamonsil TM C18 column. The mobile phase consisted of acetonitril-0. 05mol? L-1 sodium biphosphate solution ( 45∶ 55) with detection wavelength at 440nm and flow rate at 1. 0mL? min-1. RESULTS: The calibration curve of dracorhodin was linear within the range of 0. 122～ 0. 854? g ( r=0. 999 9) , with average recovery at 101. 4% ( RSD=0. 91% ) . CONCLUSION: This method is simple, reliable and reproducible, and suitable for the quality control of Diedaqili Tablets.