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RESUMEN Objetivos: Determinar el efecto citotóxico y genotóxico in vitro del extracto crudo y etanólico del rizoma de Curcuma longa L. Materiales y métodos: El efecto citotóxico fue evaluado utilizando líneas celulares DU-145, HT-29, 3T3 BALB/c. Se hallaron los porcentajes de crecimiento en 48 horas y se determinó la concentración inhibitoria 50 (CI50). El efecto genotóxico en el ADN genómico humano se determinó mediante el método Tomasevich. Resultados: El extracto crudo produjo una CI50 de 12,98 ± 0,21 μg/mL para la línea celular tumoral HT-29, que es inferior a DU-145 con una CI50 de 36,77 ± 9,12 μg/mL; el extracto etanólico presentó una CI50 de 13,24 ± 0,77 y 20,54 ± 2,58 µg/mL para ambas líneas celulares, respectivamente; el compuesto estándar curcumina presentó una CI50 de 3,96 ± 0,60 y 13,94 ± 2,79 μg/mL, respectivamente. El extracto crudo a concentraciones de 50 y 100 mg/mL fragmentó entre el 40% a 95% de ADN genómico humano; mientras que, a 200 mg/mL, la fragmentación fue mayor al 95%. El extracto etanólico a todas las concentraciones no fragmentó el ADN. La curcumina a 200 mg/mL fragmentó menos del 5% de ADN genómico humano. Conclusiones: Los extractos crudo y etanólico de Curcuma longa L. demuestran efecto citotóxico in vitro diferencial para la línea celular tumoral humana DU-145 y HT29 semejante al compuesto estándar curcumina. El extracto crudo de Curcuma longa L. presenta una potente actividad genotóxica in vitro frente al ADN genómico humano, esta actividad está ausente en el extracto etanólico.
ABSTRACT Objectives: To determine the in vitro cytotoxic and genotoxic effect of the crude and ethanolic extract from the Curcuma longa L. rhizome. Materials and methods: The cytotoxic effect was evaluated using DU-145, HT-29, 3T3 BALB/c cell lines. The growth percentages in 48 hours; and the half maximal inhibitory concentration (IC50) were determined. The genotoxic effect on human genomic DNA was determined using the Tomasevich method. Results: Crude extract produced an IC50 of 12.98 ± 0.21 μg/mL for the HT-29 tumor cell line, which is lower than the value obtained for DU-145, with an IC50 of 36.77 ± 9.12 μg/mL. The ethanolic extract presented an IC50 of 13.24 ± 0.77 and 20.54 ± 2.58 μg/mL for both cell lines, respectively; the curcumin standard compound presented an IC50 of 3.96 ± 0.60 and 13.94 ± 2.79 μg/mL, respectively. Crude extract concentrations of 50 and 100 mg/mL fragmented between 40% to 95% of human genomic DNA; while at 200 mg/mL, fragmentation was greater than 95%. The ethanolic extract at all concentrations did not fragment the DNA. Curcumin at 200 mg/mL fragmented less than 5% of human genomic DNA. Conclusions: The crude and ethanolic extracts of Curcuma longa L. demonstrate different in vitro cytotoxic effects for the human tumor cell lines DU-145 and HT-29; similar to the standard curcumin compound. The crude extract of Curcuma longa L. shows a potent genotoxic in vitro activity against human genomic DNA; this type of effect is not produced by the ethanolic extract.
Subject(s)
In Vitro Techniques , Curcuma , Rhizome , Cell Line, Tumor , Complex Mixtures , Cell Line , HT29 Cells , Inhibitory Concentration 50 , BALB 3T3 CellsABSTRACT
Objective To determine the therapeutic efficacy of curcumine on inflammatory bowel disease(IBD)and its underlying mechanisms.Methods The rat ulcerative colitis model was developed by using 30 g/L dextran sulfate sodium.Twenty-four young rats were divided into 4 groups:normal group,model group,curcumin 100 mg/kg group(curcumin 100 group),and curcumin 300 mg/kg group(curcumin 300 group).Curcumin was given through gastric gavage once a day for 7 days.General condition,disease activity index(DAI)and histopathological score(HPS)were evaluated.The proportions of CD4+ CD25+ Foxp3+ regulatory T cells(Treg)and CD4+ IL-17+ helper T lymphocyte 17(Th17)in CD4+ T cells in rats,and spleen cells were calculated by using flow cytometry.Immunohistochemical method was performed to determine the level of SOCS3 in colon tissues.Results After curcumin administered through gastric gavage for 5 days,compared with the DAI in the model group[(7.50±0.32)scores],HPS in the curcumin 100 group[(4.00±1.10)scores] and the curcumin 300 group [(5.00±0.89)scores] significantly reduced,and the difference was significant(F=12.469,P0.05).Compared with the model group[(6.5±1.5)％],the propotion of CD4+ CD25+ Foxp3+ Treg cells in CD4+ T cells in the curcumin 100 group[(9.9±1.0)％],the curcumin 300 group [(9.3±0.7)％] and the normal group[(9.6±1.1)％]was obviously upregulated(F=16.635,P0.05).Compared with the model group[(3.5±1.4)％],the propotion of CD4+ IL-17+ Th17 cells in CD4+ T cells in the curcumin 100 group[(2.0±0.9)％],the curcumin 300 group [(1.2±0.6)％] and the normal group[(2.0±0.9)％] was downregulated(F=5.155,P0.05).There was no obvious difference between the curcumin 100 group and the curcumin 300 group in these indicators respectively(all P>0.05).Conclusions Curcumin probably attenuates IBD severity,reduces inflammation and regulates the balance of Treg/Th17 through the inhibition of SOCS3 expression.
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Objective: To establish a method for the determination of curcumine and berberine hydrochloride in Shangke Dieda paste. Methods:An HPLC method was adopted to determine the content of curcumine and berberine hydrochloride in Shangke Dieda paste. For curcumine, the column was InertSutain C18 (250 mm × 4. 6 mm,5 μm); the mobile phase was acetonitrile and 4% acetic acid solution (44 ∶56);the flow rate was 1. 0 ml·min-1;the column temperature was 25℃;the detection wavelength was 430 nm;the sample size was 10μl. For berberine hydrochloride, the column was InertSutain C18 (250 mm × 4. 6 mm,5μm);the mobile phase consisted of acetonitrile-0. 1% phosphoric acid (44 ∶56, 0. 2 g dodecyl sodium sulfate was added to 100 ml solution); the flow rate was 1. 0 ml·min-1;the column temperature was 25℃ ;the detection wavelength was 345 nm;the sample size was 10 μl. Results:A good linear correlation was obtained within the range of 0.01-0.50 μg (r =0.999 3) for curcumin and 0.02-0.16 μg(r =0. 999 9) for berberine hydrochloride. The average recovery was 101. 03% with RSD of 1. 75% for curcumin and 99. 20% with RSD of 0. 64% for berberine hydrochloride (n=9). Conclusion:The established method is simple, accurate, sensitive and specific, which can be used for the quality control of Shangke Dieda paste.
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Background Proliferative vitreoretinopathy (PVR) is a common cause of vision loss clinically,and retinal pigment epithelium (RPE) cells play a major part in this disease.Studying the effect of traditional Chinese medicine on RPE cells are of great importance to reveal the pathogenesis and prevention of PVR,which were rarely reported.Objective This study was to study and compare the inhibition effect among curcumin,salvia miltiorrhiza and matrine on IL-1β-induced proliferation of rabbit RPE cells.Methods RPE cells at passages 3-4 were enrolled for the research and identified by transmission electron microscope.The proliferation effect of IL-1 β (2.5,5.0,10.0,20.0 μg/L) and inhibitory effect of curcumin (5,10,20 μg/ml),salvia miltiorrhiza (5,10,20 μg/ml)or matrine (100,200,400 μg/ml) on RPE cells 24,48 and 72 hours after cultivation were studied by MTT assay.The 50％ inhibitory dose (IC50) of the three medicines were analyzed by regression analysis.The use and feeding of the experimental animals were followed by the ARVO Statement.Results RPE cells isolated from the rabbit eye were in round shape and abundant in melanin;The melanin significantly decreased in the fourth generations of RPE cells.Immunohistochemistry showed that the RPE cells was positive for keratin (AE1/AE3).The proliferation rates of RPE cells were statistically different among different concentrations of IL-1β 24,48 and 72 hours after cultivation (Ftime =30.33,P =0.00;Fconcentration =9.37,P =0.00);The proliferation rates of RPE were significantly different among different time points or different concentrations of IL-1β (all at P ＜ 0.05).And the proliferation rate run up to maximum at 10 μg/L after 72 hours of cultivation.The inhibitory rates of the three medicines were statistically different among different time points or different concentrations (curcumin:Ftime =128.75,P =0.00;Fconcentration =334.05,P=0.00.salvia miltiorrhiza:Ftime =39.32,P=0.00;Fconcentration =165.57,P=0.00.matrine:Ftime =267.76,P =0.00;Fconcentration =912.34,P =0.00).The three medicines dose-dependently and time-dependently inhibit IL-1β-induced proliferation of RPE cells,with significant differences between the adjacent time points and concentrations (all at P＜0.05).The IC50 were 26.77,19.01 and 9.45 μg/ml for curcumin;33.72,23.47 and 12.56 μg/ml for salvia miltiorrhiza,570.96,352.25 and 97.50μg/ml for matrine 24,48 and 72 hours after cultivation.Conclusions The proliferation of RPE cells can be stimulated by IL-1β,and the maximal proliferation occurred with a concentration of 10.0 μg/L IL-1β.Curcumin,salvia miltiorrhiza and matrine dose-dependently and time-dependently inhibit proliferation of RPE cells induced by IL-1β.Curcumin is the best medicine to inhibit the proliferation of RPE cells.
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Objective: To design and synthesize conjugates of alkoxylbiphenyl/C5-curcumine with antitumor activities. Methods: Bicyclol, acetone and benzaldehyde with various substituents were used as raw materials. The target compounds were obtained by oxidizing and Aldol condensation reaction. And their antitumor activities were evaluated by MTS assay in the parental sensitive K562 and drug-resistant K562/A02 cell lines. Results: Twelve alkoxylbiphenyl/C5-curcumine conjugates were prepared.
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Objecti ve To design and synthesize conjugates of alkoxylbiphenyl/C5-curcumine with antitumor activities. Methods Bicyclol,acetone and benzaldehyde with various substituents were used as raw materials. The target compounds were obtained by oxidizing and Aldol condensation reaction. And their antitumor activities were evaluated by MTS assay in the parental sensitive K562 and drug-resistant K562/A02 cell lines. Results Twelve alkoxylbiphenyl/C5-curcumine conjugates were prepared. Conclusion The synthetic procedure of the target compounds is simple,and the yield is high. Compound 3b could significantly inhibit K562 and drug-resistant K562/A02 cell proliferation,and the IC50 is 5.38 and 3.60μmol/L,respectively.
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OBJECTIVE:To prepare Curcumine solid lipid nanoparticles(Cur-SLN).METHODS:Cur-SLN was prepared by emulsification evaporation-solidification at low temperature.The formulation of Cur-SLN was optimized by orthogonal experiment with the contents of curcumine,poloxamer 188,docosanoic acid glyceride lecithin used as factors and with entrapment efficiency as index;meanwhile,the appearance,particle size,surface potential,entrapment efficiency and the release profile of curcumine in vitro were studied.RESULTS:The optimized preparation conditions were as follows:the contents of curcumine,poloxamer,docosanoic acid glyceride lecithin were 8 mg,320 mg,140 mg and 320 mg,respectively.The Cur-SLN assumed spherical shape with particle size at(134?5)nm,entrapment efficiency at(67.4?1.3)% and mean surface potential at(-23.5?1.5)mV.The release profile of curcumine in vitro fitted Higuchi equation.CONCLUSION:It is feasible to prepare Cur-SLN by emulsion evaporation-solidification at low temperature,and the study results serve as experimental evidence for developing new curcumine injection.
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Objective To explore the effect of curcumine on the proliferation and heparanase expression in blood vessel endothelium cells isolated from ovarian cancer tissues. Methods Blood vessel endothelium cells were obtained from pathologically identified ovarian cancer tissues by primary culture. The control group was the normal blood vessel endothelium cells. The experimental groups was treated with curcumine at 2,10,50 and 250 ?g/ml for 24,48 h and 72 h respectively. The cytostasis rate and cell cycle changes of the primarily cultured cells were detected by MTT assay and flow cytometry respectively. The transcriptional and translation levels of heparanase were detected by RT-PCR and immunofluorescent staining. The MDA content and LDH activity in the supernatant were detected too. Results In the curcumine groups,the cell proliferation was inhibited in a dose-dependent and time-dependent manner ( P
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Objective To explore the effect of curcumine on rat cerebral ischemia model of blood stasis.Methods The blood-stasis rat model was established by injection of dexamethasone,then was given gastric infusion curcumine suspension for 10 continuous days.One hour after administration on the 11th day,we made ligation of the bilatory common carotid arteries of rats for 30 min to induce cerebral ischemia rat model.The rat blood sample was used for the detection of whole blood viscosity,and the brain was taken out for the observation of cerebral homogenate ATP and the pathological changes of brain tissue.Results The blood-stasis rats model was established successfully by intramuscular injection of dexamethasone,and cerebral ischemia rats model was established successfully by ligation of bilateral common carotid artery.Compared with the model group,curcumine could significantly reduce the whole blood viscosity of blood-stasis rats model of cerebral ischemia,and significantly increase the activity of Na+-K+-ATP,Mg2+ATP,Ca2+-ATP and Ca2+-Mg2+-ATP in brain homogenate,significant relieve the atrophy of nerve cells and glial cells of the model rats.Conclusion Curcumine can significantly improve the whole blood viscosity,ATP activity in the brain homogenate and morphological changes of brain tissue in rats model of blood stasis.
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Objective To investigate the effect of curcumine on the expression of matrix metalloproteinase-9 (MMP-9) in human umbilical vein endothelial cell (HUVEC). Methods HUVEC were cultured and identified in vitro .After cultured with curcumine at different concentration for 24 hours, the expression of MMP-9 in HUVEC was detected by cytoimmunochemical staining method and ELISA . Results All concentrations of curcumine can restrain the expression of HUVEC after co- culturing with the cells for 24 hours. The inhibitory effect got stronger with the increase of the concentration of curcumine. Conclusion The curcumine can depress the expression of matrix metalloproteinase-9 in HUVEC obviously.
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OBJECTIVE:To develop an HPLC method for the determination of the content of curcumine in Rujiexiao tablets.METHODS:The separation of sample was performed on VP-ODS C18 column (250 mm?4.6 mm,5 ?m).The mobile phase consisted of acetonitrile-0.2% phosphoric acid(61∶39) with a flow rate of 1.0 mL?min-1.The detection wavelength was set at 428 nm. RESULTS:The calibration curve of curcumine was linear within the range of 0.036～0.216 ?g (r=0.999 8) with an average recovery of 98.93%(RSD=0.99%,n=9).CONCLUSION:This method was proved to be simple,reliable and reproducible,and applicable for the quality control of Rujiexiao tablets.