ABSTRACT
The Chinese medicine injections prepared by the natural products containing sesquiterpenoids caused various adverse reactions in clinical use, among which skin allergic reactions are the most common. However, whether the reason of allergic reaction was related to the three isoprene units contained in the sesquiterpenoids is not clear, so the evaluation of drug safety has important guiding significance. The sesquiterpenoids are small molecular substances, and they are not antigens or haptens. They may induce anaphylaxis reactions by acting mast cells directly. Current research confirmed that Mas-related G protein-coupled receptor-X2 (MRGPRX2) which is a 7-transmembrane G protein coupled receptor on mast cells was a key target mediated allergic reactions induced by many small molecular drugs. Unlike IgE-mediated allergic reactions, pseudo-allergic reaction is related to dosage and dosing rate, and occurs in the first exposure to the sensitizer. In this paper, a series of experiments in vitro found that not all sesquiterpenoids caused anaphylactoid reactions. Ginsenoside Re, ginsenoside Rb1 and germacrone were selected as representative of sesquiterpenoids for calcium imaging assay. The data confirmed that only germacrone activated calcium mobilization through MRGPRX2, causing an increase in intracellular calcium ion concentration in mast cells. Furthermore, the release rate of β-hexosaminidase and the release amount of histamine analysis confirmed that germacrone induced mast cells degranulation directly. Knockdown of MRGPRX2 expression by siRNA and competitive binding experiments against ciprofloxacin were used to prove the target of germacrone was MRGPRX2. The results indicated that germacrone could activate mast cells directly to induce anaphylactoid reaction via MRGPRX2, which might be the reason of skin allergic reactions caused by injections containing germacrone.
ABSTRACT
Objective:To establish the gray relational analysis for quality evaluation of the samples of Curcumae Radix introduced in Zhongshan. Method:With volatile oil and curcumin as Q-markers,and alcohol extract,germacrone,germacr-1(10)-ene-5,8-dione and curcumin as comprehensive evaluation index, the contents of the four main components in 72 samples of Curcumae Radix of 3 different varieties introduced in Zhongshan from 3 different regions were determined. The grey relational method was used to build the gray correlation evaluation model for Curcumae Radix introduced in Zhongshan. Result:The relative correlation degree (γi) of 72 samples was between 0.262 and 0.697,in which γi was above 0.450 for 10 samples,and below 0.300 for 37 samples,indicating great differences in the quality of Curcumae Radix after introduction. The γi was 0.697 and 0.525 respectively for No.MY-W-4 and No.MY-W-1 from Curcumae Radix in Mayu with the best quality. The average values of γi for the samples of 3 different varieties from 3 different regions were between 0.281 and 0.420,and Mayu samples had the maximum average value,indicating that Mayu samples had the highest overall quality of,and could be introduced as excellent resources. Conclusion:The evaluation method combined with GRA method and multi-index quantification was simple,objective and comprehensive, and could be used to evaluate the quality of Curcumae Radix introduced in Zhongshan,so as to provide references for screening high-quality provenance.
ABSTRACT
OBJECTIVE:To study the distribution characteristics of germacrone from zedoary turmeric oil (ZTO) in each tis-sue of mice,and to provide reference for further application of zedoary turmeric oil. METHODS:30 KM mice were given zedoary turmeric oil 0.5 mL;6 mice were randomly selected 1,2,4,8,12 h after medication,respectively. The contents of germacrone in heart,liver,spleen,lung and kidney tissues were determined by HPLC. 15 KM mice were selected,medication and sampling method were same as above;3 mice were collected at each time point respectively. The fluorescence intensity of germacrone in above sections were observed by fluorescence. The same number of mice were selected as control in 2 trials. RESULTS:The con-centration of germacrone in each tissue 1-4 h increased gradually as time and reached the peak value at 4 h. The contents of ger-macrone in liver and spleen were significantly higher than in heart and lung. The concentrations of germacrone in each tissue were ranked as liver>spleen>kidney>heart>lung. The results of fluorescence intensity observation was same as above results. CON-CLUSIONS:Results of 2 methods show same distribution characteristics of germacrone in mice tissues,and indicate that ger-macrone is distributed more in liver,spleen and kidney tissues and less in heart and lung.
ABSTRACT
OBJECTIVE: To establish a method for the simultaneous determination of curcumin and germacrone in Curcuma wenyujin by ultrasound-assisted ionic liquid-reversed phase liquid chromatography. METHODS: An Agilent ZORBAX Eclipse Plus C18 column (4. 6 mm × 250 mm, 5 μm) was used. Gradient elution was conducted with the mobile phase consisting of acetonitrile and 0.1% phosphoric acid at the flow rate of 1.0 mL·min-1. The detection wavelength was set at 265 nm, the flow rate was set at 1.0 mL·min-1, and the column temperature was maintained at 30 ℃. RESULTS: The result showed that curcumin and germacrone had the highest extraction yield when the [BMIM]PF6 methanol solution concentration of 0.6 mol·L-1 was used as the extraction solvent and the solid-liquid ratio was 1:20. Under the optimized condition, good linearity of curcumin was shown for curcumin in the range of 0.041 2-2.06 μg (r=0.999 9)and the average recovery was 98.31%. The good linearity of germacrone was shown for germacrone in the range of 0.010 04-0.502 μg (r=0.999 8)and the average recovery was 97.62%. CONCLUSION: This method is convenient, environmentally friendlygreen and highly reproducible. It is beneficial important to study the extraction method of the active gradients of the traditional Chinese medicine Curcuma wenyujin active gradient and its quality control of Curcuma wenyujin.
ABSTRACT
This study focuses on the protective effect of germacrone on human umbilical vein endothelial cells(HUVECs) damaged by H2O2-induced oxidative stress and its possible mechanisms. The oxidative damage model was established by using 500 μmol•L⁻¹ H2O2 to treat HUVECs for 3 hours, and then protected with different concentrations of germacrone for 24 hours. The effect of germacrone on cell viability of HUVECs damaged by H2O2 was detected by MTT. The contents of PGI2, TXB2, ET-1, t-PA, PAI-1, TNF-α and IL-6 were detected by ELISA. The content of NO was detected by using nitrate reductase method. Colorimetry was used to detect NOS and GSH-Px. The contents of MDA, SOD and LDH were detected by TBA, WST-1 and microplate respectively. Apoptosis was observed by Hoechst 33258 fluorescent staining. The mRNA expressions of Bax, Bcl-2 and Caspase-3 in cells were detected by RT-PCR. The results showed that the cell damage rate was 52% after treated with 500 μmol•L⁻¹ H2O2 for 3 hours. The cell activity was increasing with the rise of germacrone concentration within the range of 20-200 mol•L⁻¹. Compared with normal group, the contents of PGI2, NO, T-NOS, t-PA, SOD, GSH-Px and Bcl-2 mRNA expressions were lower after damaged with H2O2. The contents of PAI-1, ET-1, IL-6, TNF-α, TXB2, LDH, MDA, Bax mRNA and Caspase-3 mRNA expressions were increased. Compared with model group, the contents of PGI2, NO, T-NOS, t-PA, SOD, GSH-Px and Bcl-2 mRNA expressions were increased after treated with germacrone. The contents of PAI-1, ET-1, IL-6, TNF-α, TXB2, LDH, MDA, Bax mRNA and Caspase-3 mRNA expressions were lower after treated with germacrone. According to Hoechst 33258 fluorescence staining, compared with normal group, the cell membrane and the nucleus showed strong dense blue fluorescence, and the number of cells significantly decreased in model group. Compared with model group, blue fluorescence intensity decreased in drug group. The above findings demonstrate that germacrone may improve the effect on HUVECs damaged by H2O2-induced oxidative stress by resisting oxidation and inhibiting cell apoptosis.
ABSTRACT
OBJECTIVE:To establish a method for the simultaneous determination of curdione,germacrone and furanodiene of zedoray turmeric oil in Ruxiankang injection. METHODS:HPLC was performed on the column of Zorbax SB-C18 with mobile phase of acetonitrile-water (gradient elution) at a flow rate of 1.0 ml/min,detection wavelength was 216 nm,column temperature was 30℃,and the injection volume was 10 μl. RESULTS:The linear range was 60-480 μg/ml for curdione(r=0.999 0),40-320 μg/ml for germacrone(r=0.999 0)and 40-320 μg/ml(r=0.999 0);RSDs of precision,stability and reproducibility tests were lower than 2.0%;recoveries were 95.21%-99.89%(RSD=1.6%,n=6),102.33%-104.89%(RSD=1.0%,n=6)and 97.38%-99.06%(RSD=0.7%,n=6),respectively. CONCLUSIONS:The method is simple and accurate,and can be used for simultaneous contents deter-mination of curdione,germacrone and furanodiene of zedoray turmeric oil in Ruxiankang injection.
ABSTRACT
Abstract The gorgonian Phyllogorgia dilatata Esper is an octocoral known to be source of biologically active terpenes. In this study, odoriferous compounds present in P. dilatata tissues were investigated, due to their exotic olfactory notes. The search of volatile compounds was performed in a dichloromethane/methanol extract submitted to a silica gel vacuum chromatography and HPLC, yielding the isomers (Z,E) and (E,E)-germacrones, identified by GC/MS, 1 and 2D NMR. The stereochemistry of (E,E)-germacrone, as well as its preferred conformation, was confirmed by NOESY. Sensory analysis of the two isomers revealed a fragrant, citrus, woody and weak marine odor, similar to the odor of the natural gorgonian, and (E,E)-germacrone has a three times more intense aroma than the (Z,E) isomer.
ABSTRACT
Objective To establish the chromatographic fingerprint of Curcuma wenyujin by HPLC.Methods The chromatographic fingerprint of C.wenyujin was established by HPLC with Shimadzu VP-ODS C18 column(250 mm?4.6 mm,5 ?m),acetonitrile-water(0.1 phosphoric acid) gradient elution,the flow rate of 1.0 mL/min,column temperatures of 30 ℃,and detective wavelength at 210 nm.ResultsTaking germacrone as the reference peak,14 common-peaks were selected as the fingerprint peaks.The similarities among samples of C.wenyujin among 19 approved samples and their standard chromatographic fingerprints were calculated with a software "Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica" published by GPC(Version 2004A) and cluster analysis.ConclusionThis method with better accuracy and reproducibility could be used in the quality control of C.wenyujin.
ABSTRACT
AIM: To establish a HPLC method for determination of curcumol and germacrone in Zedoary Turmeric Oil and Glucose Injections. METHODS: RP-HPLC was applied for quantitative analysis of curcumol and germacrone. Hypersil ODS2 column (250 mm? 4.6 mm, 5 ?m) and gradient elution was used. Mobile phase A was acetonitrile and mobile phase B was water. Phase A time-percentage composition was as follows:0-3 min,45%;3-30 min,45%→65%;30-38 min, 65%; 38-45 min, 65%→90%;45-55 min,90%→100%;55-65 min,100%. The flow rate was at 1.0 mL?min -1 . Diode array detector was set at 214 nm and column temperature was at 25 ?C. RESULTS: The calibration curves of curcumol and germacrone were linear in the range of 0.26 - 6.5 ?g(r= 0.999 9 ) and 0.112 5 - 2.812 5 ?g(r=1), The average recoveries of them were 102.1% and 98.8% (n=5), respectively. CONCLUSION: Curcumol and germacrone can be determined by the method, and this will improve the quality control of Zedoary Turmeric Oil and Glucose Injections.
ABSTRACT
OBJECTIVE:To determine the content of germacrone in rhizoma curcuma by HPLC. METHODS: C18 was the chromatographic column, the mobile phase was acetonitrile and water(gradient elution) with a flow rate of 1.0ml/min, the detection wavelength was at 214nm. RESULTS:The linear range of germacrone was 0.95～285?g/ml (r=0.9 999),the mean recovery rate was 97.89%(RSD=2.52%,n=5).CONCLUSION:The method is simple and reliable and which can be used for the content determination and the quality control of rhizoma curcuma.