Parida Yamada

Induction of growth inhibition and differentiation of human leukemia HL-60 cells by a tunisian gerboui olive leaf extract

Cancer protection associated with the consumption of olive products is well established, but not for leukemia. The protective effects of olive (Olea europaea L.) leaves were investigated by incubating human promyelocytic leukemia HL-60 cells with olive leaf extracts (OLEs) from seven principal Tunisian olive varieties, namely, Chemchali, Chemlali, Chétoui, Gerboui, Sayali, Zalmati and Zarrazi. The results showed significant growth inhibition of HL-60 cells incubated for 48 h with a 100-fold dilution of each OLE which had been obtained by incubating 10 g of dried leaves in 100 ml of 70% ethanol for one week with subsequent ultrafiltration. DNA fragmentation was observed in the cells incubated for 19 h with a 100-fold dilution of the Chemchali, Chemlali and Zalmati extracts. The results of a nitroblue tetrazolium (NBT) assay revealed NBT reduction, a differentiation marker, by the OLE-treated cells after an overnight incubation. The Gerboui extract showed the highest NBT reduction ability at more than 90%. An HPLC analysis revealed the presence of apigenin 7-glucoside in the extract, which was found in subsequent experiments to be responsible for the Gerboui extract-mediated cell differentiation.

Inhibitory Effect of Fulvic Acid Extracted from Canadian Sphagnum Peat on Chemical Mediator Release by RBL-2H3 and KU812 Cells

Fulvic acid (FA) was extracted and purified from Canadian Sphagnum peat (CP-FA) and characterized by using an element analysis meter, Fourier transform infrared (FT-IR) spectroscopy, electron spin resonance (ESR) spectroscopy, and 13C-nuclear magnetic resonance (13C-NMR) spectroscopy. To investigate the antiallergic effect of CP-FA, we incubated rat basophilic leukemia (RBL-2H3) cells with 0.001–10.0 μg/ml of CP-FA and determined the β-hexosaminidase release inhibition at different response stages. The intracellular calcium [Ca2+] i level was also determined by using Fluo 3-AM, a calcium-specific fluorescent probe, and the cytotoxicity of CP-FA was determined by the 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. The results revealed that RBL-2H3 cells incubated for 48 h with 0.001–10.0 μg/ml of CP-FA did not show any decreased viability. CP-FA inhibited the β-hexosaminidase release by IgE-sensitized, antigen-stimulated RBL-2H3 cells at the antigen-antibody binding stage and the antibody-receptor binding stage. CP-FA also inhibited histamine release from A23187 plus PMA- or compound 48/80-stimulated KU812 cells. Furthermore, there was a decrease in the intracellular [Ca2+] i level in IgE-sensitized cells incubated with CP-FA and stimulated with antigen. Our results show that CP-FA may be useful for the treatment or prevention of allergic diseases.

Inhibitory effect of various Tunisian olive oils on chemical mediator release and cytokine production by basophilic cells.

Tunisian olive oils have been traditionally used as a medicinal food for chronic inflammation. To investigate the antiallergic effect of virgin olive oil samples from five principal olive varieties grown in various regions of Tunisia, we used the type I allergy reaction model using rat basophilic leukemia (RBL-2H3) cells and different dilutions of olive oil samples to determine beta-hexosaminidase release inhibition at two different response stages. Results showed that the Sayali olive oil significantly inhibited beta-hexosaminidase release by the IgE antibody-sensitized, BSA antigen-stimulated RBL-2H3 cells at the antibody-antigen binding stage. The result of our experiment shows that the anti-allergic effect of olive oil at this binding stage may be dependent on their flavone content. The Zarrazi olive oil significantly inhibited beta-hexosaminidase release at the antigen-receptor binding stage. Moreover, we investigated the effect of olive oil samples on histamine release and production of cytokines by activated human basophilic (KU812) cells. Different dilutions of Sayali olive oil dose-dependently inhibited the production of tumor necrosis factor-alpha (TNF-alpha) and interleukin-4 (IL-4), and different dilutions of Zarrazi olive oil dose-dependently inhibited histamine release and IL-4 production by calcium ionophore A23187 plus phorbol 12-myristate 13-acetate (PMA)-stimulated KU812 cells.

Hirseins inhibit melanogenesis by regulating the gene expressions of Mitf and melanogenesis enzymes

Please cite this paper as: Hirseins inhibit melanogenesis by regulating the gene expressions of Mitf and melanogenesis enzymes. Experimental Dermatology 2009; 19: 450–457.

Inhibitory Effect of Acteoside Isolated from Cistanche tubulosa on Chemical Mediator Release and Inflammatory Cytokine Production by RBL-2H3 and KU812 Cells

The immediate-type allergic reaction is involved in many allergic diseases such as asthma, allergic rhinitis, and sinusitis. In this study, we investigated the effect of acteoside extracted from CISTANCHE TUBULOSA (Schrenk) R. Wight on the basophilic cell-mediated allergic reaction. The effect of acteoside on β-hexosaminidase release and intracellular [Ca (2+)] I level from rat basophilic leukemia (RBL-2H3) cells was determined. Also, ELISA was used to determine the level of histamine, tumor necrosis factor (TNF)- α, and interleukin (IL)-4 on human basophilic (KU812) cells. The effect of acteoside on basophilic cell viability was determined using the 3-[4,5-dimethylthiazolyl]-2,5-diphenyltetrazolium bromide (MTT) assay. These results indicated that 0.1-10.0 µg/mL acteoside inhibits the release of β-hexosaminidase and [Ca (2+)] I influx from IgE-mediated RBL-2H3 cells. Moreover, acteoside inhibited histamine release, TNF- α, and IL-4 production in a dose-dependent manner from calcium ionophore A23187 plus phorbol 12-myristate 13-acetate (PMA) or compound 48/80-stimulated KU812 cells. Our findings provide evidence that acteoside inhibits basophilic cell-derived immediate-type and delayed-type allergic reactions. This is the first report describing antiallergic activity of acteoside extracted from CISTANCHE TUBULOSA on basophilic cells.

Isolation of 5-(hydroxymethyl)furfural from Lycium chinense and its inhibitory effect on the chemical mediator release by basophilic cells.

The effect of hot water extracts of LYCIUM CHINENSE fruits (LCF) on the β-hexosaminidase (β-hexo) release by IgE sensitized BSA stimulated rat basophilic leukemia (RBL-2H3) cells was investigated. The ethylacetate (EtOAc) layer of the extract has shown an inhibitory effect on β-hexo release from RBL-2H3 cells at the antigen antibody binding stage. The water (H₂O) fraction (EFW) of the chloroform (CHCl₃) extract from the EtOAc layer also inhibited β-hexo release at the same stage in a dose-dependent manner. With column chromatography preparation, proton and carbon nuclear magnetic resonance (¹H and ¹³C NMR) spectra, electron ionization mass spectrometer (EI-MS) spectra, and high-performance liquid chromatography (HPLC) analysis, the active component was determined to be 5-(hydroxymethyl)furfural (5-HMF). Thus, the 5-HMF showed an inhibitory effect on β-hexo release at the antigen-antibody binding stage and the antibody-receptor binding stage. Furthermore, 5-HMF suppressed [Ca²+] I influx in the IgE-sensitized BSA-stimulated RBL-2H3 cells. Our results show that 5-HMF may be useful for the treatment or prevention of type I allergic diseases.

Stress response of heavy metal mixture present in wastewater and leachate on heat-shock protein 47-transfected cells

Heavy metals present in water environment and hazardous sites as single compounds or mixture may drastically affect human health. In the present work, we investigated the risk assessment of wastewater effluents and leachate with a focus on three heavy metals-nickel (Ni), cadmium (Cd), and lead (Pb)-and their combined effect on mammalian cells, using Chinese hamster ovary cells transfected with the heat-shock protein (HSP) 47 promoter. The heavy metal mixture model was designed based on the concentrations of metals in wastewater effluents and leachate sampled in Tunisia. Using a ternary diagram, we investigated the stress response of the interaction model. This research indicated that the single heavy metals induced the stress response on HSP(+) cells even at concentrations lower than the local and international guidelines. Differences in water quality likely influenced the metal responses such that the organic composition of the leachate increased the stress response induced by the heavy metals exclusively, whereas the effluents included organic compounds that were able to mask the heavy metal effect. The mixture characterization discovered the key role played by the high levels of Ni or combination of Cd and Pb to induce the highest stress response following 3-h incubation. Heat-shock protein 47 has proven its effectiveness for assessing the heavy metal mixture effect even at low concentrations. Furthermore, the combination of a bioassay system with a statistical model proved extremely useful for better understanding the major contributors to the stress response of the mixture.

Inflammatory and degranulation effect of yellow sand on RBL-2H3 cells in relation to chemical and biological constituents

Recent studie pointed out that allergic diseases have increased during the Asian dust storm event (ADSE) in Japan. Daily observations and the atmospheric concentrations of yellow sand (YS) aerosol have been increasing. In this study, YS samples collected from three sites of Japan during ADSE in 2009-2010 were used. The particles were analyzed by X-ray photoelectron spectroscopy (XPS) and X-ray fluorescence-energy dispersive spectrometer (XRF-EDS). We investigate ability of YS extract on enhancing the chemical mediator release and cytokine production from rat basophilic leukemia (RBL-2H3) cells. The dust particles at Fukuoka and Tsukuba were abundant in aluminum (Al), iron (Fe), potassium (K) and titan (Ti) than those at Naha. Concentration of the trace endotoxin and Cryptomeria japonica pollen allergen (Cry j 1) were measured in YS extract. After exposure of RBL-2H3 cells to YS extract, the β-hexosaminidase (β-hex) release, tumor necrosis factor-alpha (TNF-α) production were enhanced in RBL-2H3 cells. This process depends on endotoxin, Cry j 1 and other allergen present in the YS extract. YS water extract also show a strong cytotoxic effect on the cells. This data suggest that low levels of endotoxin and Cry j 1 in YS may cause allergy during the ADSE.

Properties of fulvic acid extracted from excess sludge and its inhibiting effect on β-hexosaminidase release.

The physicochemical and biological properties of fulvic acid extracted and purified from excess sludge and solubilized excess sludge were studied. Solubilization was introduced to improve the recovery rate of fulvic acid from the sludge. The structural features of fulvic acid from excess sludge and solubilized excess sludge were characterized by using an elemental analysis, Fourier transform infrared spectroscopy and 1H-nuclear magnetic resonance spectroscopy, and were compared with fulvic acid extracted from peat which had an inhibitory effect on the type I allergy in our previous study. The results show that they had a higher aliphatic characteristic with lower oxygen group content than fulvic acid from peat, and that the aliphatic characteristic was further strengthened by the use of solubilization. The biological properties of fulvic acid from excess sludge and solubilized excess sludge showed an inhibitory effect on β-hexosaminidase release at the antigen-antibody binding stage and antigen-receptor binding stage by using rat basophilic leukemia cells.

Indoleacetic acid falcarindiol ester induces granulocytic differentiation of the human leukemia cell line HL-60.

Indoleacetic acid falcarindiol ester (compound 1) has previously been isolated and purified using an SiO2 column and ODS HPLC from an acetone extract of Japanese ivy (Hedera rhombea). Here we investigate the differentiation-inducing activity of compound 1 using the human promyelocytic leukemia HL-60 cell line. The effect of compound 1 on HL-60 cell viability and proliferation was determined at different treatment times using the 3-(4,5-dimethythiazol-2-yl)-2,5-diohenyl-2 H-tetrazolium bromide (MTT) assay and flow cytometry analysis. Also cell cycle kinetics were examined using propidium iodide staining of DNA. Cell differentiation was assessed by specific and non-specific esterase double staining assays, and by detection of the cell surface differentiation markers CD11b and CD14 using flow cytometry. The results showed HL-60 cell growth inhibition at 0.1 and 1.0 microg/mL compound 1, whereas 10 microg/mL was cytotoxic. The growth suppression induced by compound 1 was accompanied by G0/G1 phase arrest in the cell cycle at 1.0 microg/mL. Moreover, staining and immunochemical analysis indicated that compound 1 induced granulocytic differentiation in HL-60 cells. This is the first report describing granulocytic differentiation activity of a falcarindiol derived polyacetylenic compound on leukemia cells.