An investigation on the possible role of melatonin in melanogenesis

The possible role of melatonin in melanogenesis was investigated by performing reactions of melatonin with peroxidase + H2O2 or H2O2 only, in the presence or absence of UV irradiation. Samples of the reaction mixtures were drawn at different times (from 15 to 480 min), the enzyme (when present) was removed by ultrafiltration and the samples so obtained were analyzed by MALDI/MS.The results show that melatonin undergoes oligomerization reaction with peroxidase + H2O2, leading to heptameric species. For high reaction times the MALDI/MS data do not show the formation of larger oligomers, but UV-vis spectroscopy indicates that the oligomerization processes proceed. The failure of MALDI-TOF in the identification of larger oligomers was related to the chemical-physical and morphological behavior of melanins.In the case of UV irradiation, the formation of species originating from the O- and O2 addition to melatonin, which activate new oligomerization channels, has been observed.     

Placental extracts induce the expression of antioxidant enzyme genes and suppress melanogenesis in B16 melanoma cells

One of the activities of placental extracts (PEs) is skin-whitening effect, but the physiological and genetic mechanism for this effect has not yet been clarified. Here, we focus on PE as a regulator of antioxidant enzyme genes. Porcine PE was prepared, and its activity was investigated in B16 melanoma cells. PE treatment decreased the melanin content of UV-irradiated B16 cells in a dose-dependent manner. PE directly reduced the enzyme activity of tyrosinase in a cell-free assay. In addition, PE treatment increased the gene expression of cytosolic superoxide dismutase (SOD-1), extracellular SOD (SOD-3) and catalase but did not affect the expression of tyrosinase. Moreover, PE protected the B16 cells from H₂O₂-induced cell death. Taken together, our data suggest that PEs could play a role not only as a suppressor of melanin synthesis but also as a regulator of antioxidant genes and might protect the skin against oxidative stress.     

The role of chalcones: helichrysetin,xanthohumol, and flavokawin-C in promoting neurite outgrowth in PC12 Adh cells

Chalcones are a group of compounds widely distributed in plant kingdom. The aim of this study was to assess the neurite outgrowth stimulatory activity of selected chalcones, namely helichrysetin, xanthohumol and flavokawin-C. Using adherent rat pheochromocytoma (PC12 Adh) cells, the chalcones were subjected to neurite outgrowth assay and the extracellular nerve growth factor (NGF) levels were determined. Xanthohumol (10 μg/mL) displayed the highest (p < 0.05) percentage of neurite-bearing PC12 Adh cells and the highest (p < 0.05) NGF level in the culture medium of xanthohumol-treated cells. While, helichrysetin induced a moderately high numbers of neurite-bearing cells, flavokawin-C did not stimulate neurite outgrowth. This work supports the potential use of xanthohumol as a potential neuroactive compound to stimulate neurite outgrowth.     

A mass spectrometric investigation on the possible role of tryptophan and 7-hydroxytryptophan in melanogenesis

The activity of tyrosinase and peroxidase + H2O2 in promoting melanogenesis from tryptophan (Trp) and 7-hydroxytryptophan (7-HTP) has been investigated. The reaction samples have been drawn at different reaction times and analysed by MALDI mass spectrometry. The data obtained showed that tryptophan undergoes, under tyrosinase and peroxidase action, an oligomerization process mainly due to the reaction of anthranilic acid (AA) and Trp. However, analysing the UV and fluorescence data, it is seen that the oligomers cannot belong to the melanin pattern, but their possible role in melanogenesis is not to be excluded. Once it reacts with the two enzymes, 7-hydroxytryptophan leads to dark brown products, indicating its possible role in melanin production. In contrast to what was observed in the case of 5-hydroxytryptophan, for which oligomers were constituted by 5-hydroxytryptophan (5-HTP) and 5-hydroxytryptamine (5-HT) units, the MALDI data indicate a sharply different behaviour for 7-HTP. In fact, in the case of 5-hydroxytryptophan, oligomerization takes place through the formation of 5-hydroxytryptamine and the oligomerization products are due to mixed 5-HTP-5-HT oligomers. In the case of 7-hydroxytryptophan, the formation of 7-hydroxytryptamine (7-HT) is also observed, but it does not seem to play any role; the only oligomerization products formed are due to the reaction of 7-hydroxytryptophan and AA. The data so obtained indicate that 7-hydroxytryptophan acts like an effective melanin precursor in the presence of both tyrosinase and peroxidase + H2O2.     

Effects of quercetin on mushroom tyrosinase and B16-F10 melanoma cells

In searching for tyrosinase inhibitors from plants using L-3,4-dihydroxyphenylalanine (L-DOPA) as a substrate, quercetin was found to be partially oxidized to the corresponding o-quinone under catalysis by mushroom tyrosinase (EC 1.14.18.1). Simultaneously, L-DOPA was also oxidized to dopaquinone and both o-quinones were further oxidized, respectively. The remaining quercetin partially formed adducts with dopaquinone through a Michael type addition. In general, flavonols form adducts with dopaquinone as long as their 3-hydroxyl group is free. Quercetin enhanced melanin production per cell in cultured murine B16-F10 melanoma cells, but this effect may be due in part to melanocytotoxicity. The concentration leading to 50% viable cells lost was established as 20 microM and almost complete lethality was observed at 80 microM.     

Effect of Korean Red Ginseng and Rg3 on Asian Sand Dust-Induced MUC5AC,MUC5B,and MUC8 Expression in Bronchial Epithelial Cells

Korean Red ginseng (KRG), commonly used in traditional medicine, has anti-inflammatory, anti- oxidative, and anti-tumorigenic properties. Asian sand dust (ASD) is known to aggravate upper and lower airway inflammatory responses. BEAS-2B cells were exposed to ASD with or without KRG or ginsenoside Rg3. Mucin 5AC (MUC5AC), MUC5B, and MUC8 mRNA and protein expression levels were determined using quantitative RT-PCR and enzyme-linked immunosorbent assay. Nuclear factor kappa B (NF-κB), activator protein 1, and mitogen-activated protein kinase expression and activity were determined using western blot analysis. ASD induced MUC5AC, MUC5B, and MUC8 mRNA and protein expression in BEAS-2B cells, which was significantly inhibited by KRG and Rg3. Although ASD-induced mucin expression was associated with NF-κB and p38 mitogen-activated protein kinase (MAPK) activity, KRG and Rg3 significantly suppressed only ASD-induced NF-κB expression and activity. KRG and Rg3 inhibited ASD-induced mucin gene expression and protein production from bronchial epithelial cells. These results suggest that KRG and Rg3 have potential for treating mucus-producing airway inflammatory diseases.     

Na_6PMo_(11)FeO_(40)对小鼠黑色素瘤B16细胞内黑色素合成抑制及抑菌活性研究

研究了过渡金属取代的Kiggen型结构的杂多酸盐Na_6PMo_(11)FeO_(40)对体外培养的小鼠黑色素瘤B16细胞的生物学效应,并分析了其对4种细菌的抗菌活性.结果显示:不同浓度的Na_6PMo_(11)FeO_(40)对B16细胞的细胞形态和数量有不同程度的影响,对B16细胞的增殖率、酪氨酸酶活性以及黑色素合成量的抑制呈现出明显的浓度效应.浓度为50~200μmol/L的Na_6PMo_(11)FeO_(40)能显著抑制B16细胞增殖(P0.05或P0.01);浓度为200μmol/L的Na_6PMo_(11)FeO_(40)对酪氨酸酶活性抑制极显著(P0.01),IC50为166.5μmol/L;Na_6PMo_(11)FeO_(40)对藤黄八叠球菌、金黄色葡萄球菌、枯草芽孢杆菌和大肠杆菌均有抗菌作用,且对球菌的抑制效果优于杆菌.实验结果表明,Na_6PMo_(11)FeO_(40)可作为具有防腐抑菌功能的新型酪氨酸酶抑制剂.     

Inhibition effect of phenyl compounds from the Oryza sativa roots on melanin production in murine B16-F10 melanoma cells

Five phenyl compounds, vanillin (1), methyl trans-ferulate (2), trans-p-coumaric acid methyl ester (3), N-benzoyltryptamine (4), and N-(trans-cinnamoyl)tryptamine (5), were isolated from the roots of Oryza sativa L. and identified on the basis of spectroscopic data. Compounds 3 and 5 showed strong inhibition effect on melanin production in murine B16-F10 melanoma cells and tyrosinase activity. Also, the quantitative analysis of the compounds was carried out using LC/MS/MS experiment. Compounds 3 and 5 could be used as skin-whitening agents.     

H6[P2Mo18O62]的钒取代多金属氧酸盐对小鼠黑色素瘤细胞B16黑色素生成的调控及其机制研究

合成了五种多金属氧酸盐. 以B16小鼠黑色素瘤细胞为模型, 用噻唑蓝(MTT)法检测细胞存活率, 酶标法测定抗氧化活性和内黑色素含量及酪氨酸酶活性, 最后用分子对接模拟多金属氧酸盐和酪氨酸酶结合的机制. 研究结果表明, 两种磷钼酸(H₇[P₂Mo₁₇VO₆₂]和H₈[P₂Mo₁₆V₂O₆₂])是高效的黑色素生成抑制剂, 在200 μmol/L的浓度下对黑色素合成的抑制率为74.40%和75.14%, 对细胞内酪氨酸酶活性的抑制率为35.71%和40.00%, 随着钒原子取代个数的增加, 两种抑制活性均逐渐降低. H₇[P₂Mo₁₇VO₆₂]和H₈[P₂Mo₁₆V₂O₆₂]对细胞没有毒性. 两种多酸都有较好的1,1-二苯基-2-三硝基苯肼(DPPH)清除能力, IC₅₀分别为1.683和2.800 mg/mL. 分子对接分数低于–146 kJ/mol. 综上所述, H₇[P₂Mo₁₇VO₆₂]和H₈[P₂Mo₁₆V₂O₆₂]能抑制B16细胞黑色素的生成, 其机制与抑制酪氨酸酶的活性有关.     

Inhibitory effect of capsaicin on B16-F10 melanoma cell migration via the phosphatidylinositol 3-kinase/Akt/Rac1 signal pathway

Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide), the major pungent ingredient of red pepper, has been reported to possess anti-carcinogenic and anti-mutagenic activities. In this study, the anti-migration activity of capsaicin on highly metastatic B16-F10 melanoma cells was investigated. Capsaicin significantly inhibited the migration of melanoma cells without showing obvious cellular cytotoxicity at low doses. This effect correlated with the down-regulation of phosphatidylinositol 3-kinase (PI3-K) and its downstream target, Akt. Although B16-F10 cell migration was increased by the PI3-K activator through the activation of Akt, these PI3-K activator-induced phenomena were attenuated by capsaicin. Moreover, capsaicin was found to significantly inhibit Rac1 activity in a pull-down assay. These results demonstrate that capsaicin inhibits the migration of B16-F10 cells through the inhibition of the PI3-K/Akt/Rac1 signal pathway. The present investigation suggests that capsaicin targets PI3-K/Akt/ Rac1-mediated cellular events in B16-F10 melanoma cells. Consequently, capsaicin administration should be considered an effective approach for the suppression of invasion and metastasis in malignant melanoma chemotherapy.     

N-Alkylamides from Piper longum L. and their stimulative effects on the melanin content and tyrosinase activity in B16 melanoma cells

Abstract

Piper longum L., known as long pepper, is an edible and medicinal plant used as spice and for the treatment of stomach disease and analgesia in traditional Chinese medicine. N-Alkylamides are the major secondary metabolites in this plant. Sixteen known N-alkylamides were isolated from P. longum. Their structures were established on the basis of spectroscopic data and comparison to reported literatures. Among them, five compounds were isolated from this plant for the first time. Ethanol extract, compounds 1, 2, 3, 7 and 11 exhibited potent ability to increase the melanin content and weak stimulative effect on the tyrosinase activity in a concentration-dependent manner. Moreover, compound 2 also presented strong capacity to increase the tyrosinase activity in a concentration-dependent manner. These results indicated that P. longum might be a good natural source of lead compound for skin disorder diseases.     

Effect of substituents in the ligand on oxidation of alkanes catalyzed by binuclear oxo-bridged iron complexes

Oxidation of alkanes (methane, ethane, hexane, and cyclohexane) by hydrogen peroxide andtert-butyl hydroperoxide in acetonitrile catalyzed by binuclear -oxo-bridged iron complexes [Fe₂OL₄(H₂O)₂](ClO₄)₄ and [Fe₂OL₂PhCOO)₂(H₂O)₂](ClO₄)₂, where L = bpy, 4,4-Me₂bpy, 4,4-(ClCH₂)₂bpy, phen, and 5-NO₂phen, was studied. It was shown that the nature of the substituent in the ligand affects both the rate of the catalyzed peroxide decomposition and catalytic activity of the complexes studied in the alkane oxidation.Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 12, pp. 2518–2520, December, 1995.This work was financially supported by the Russian Foundation for Basic Research (Project No. 94-03-08529), the International Science Foundation (Grant No. REUOOO), the European Foundation INTAS (Grant No. 93-315), and Amoco Company (USA).     

Effect of replicated polymeric substrate with lotus surface structure on adipose-derived stem cell behaviors

We fabricated polystyrene substrates with lotus leaf surface structure (LLSS) and investigated cell behaviors, including attachment, morphology, proliferation, and differentiation of adipose-derived stem cells (ASCs) on them. Compared to the flat substrate, the LLSS substrate induced higher cell attachment rate, but did not significantly change the cell proliferation rate. In addition, ASCs on the LLSS substrate exhibited relatively narrower spreading morphology and less organized cytoskeleton, there by resulting in smaller sizes of cells than those on the flat substrate. According to histochemical staining and RT-PCR analysis, the LLSS substrate induced higher adipogenic differentiation of ASCs than the flat substrate, while chondrogenic and osteogenic differentiation were decreased.