海藻酸钠对酪氨酸酶的抑制作用
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摘要
通过酶抑制动力学实验考察了海藻酸钠在实验体系和黑素细胞内对酪氨酸酶和黑色素生成的抑制作用,利用荧光光谱分析海藻酸钠对酪氨酸酶抑制作用机理,同时考察海藻酸钠对黑素细胞的毒性。结果表明,在64 mmol/L浓度内,细胞活性均在90%以上,显示出良好的生物相容性。海藻酸钠对酪氨酸酶活性及黑色素产量均具有一定的抑制效果,且抑制类型在20 mmol/L浓度内呈明显的浓度依赖型。海藻酸钠与酪氨酸酶相互作用后,对其内源性荧光产生了淬灭作用。酪氨酸酶的最大发射波长出现了红移,表明海藻酸钠与酪氨酸酶相互作用时,改变了其氨基酸残基的疏水性环境。
Enzyme inhibition kinetics experiments were carried out in vitro and in melanocytes to study the inhibitory effect of sodium alginate on the activity of tyrosinase and melanin deposition. The inhibition mechanism of sodium alginate on tyrosinase was analyzed by fluorescence spectroscopy, and the toxicity of sodium alginate on melanocyte was also investigated. The results show that within the concentration of 64 mmol/L, the cell viability was above 90%, indicating good biocompatibility. Sodium alginate had a certain inhibitory effect on tyrosinase activity and melanin production, which was in a concentration-dependent manner below the concentration of20 mmol/L. The endogenous fluorescence of tyrosinase was partly quenched after interacting with sodium alginate.The redshift of the maximum emission wavelength appearing in the fluorescence spectrum of tyrosinase indicated that the interaction between sodium alginate and tyrosinase changed the hydrophobic environment of its amino acid residues.
Enzyme inhibition kinetics experiments were carried out in vitro and in melanocytes to study the inhibitory effect of sodium alginate on the activity of tyrosinase and melanin deposition. The inhibition mechanism of sodium alginate on tyrosinase was analyzed by fluorescence spectroscopy, and the toxicity of sodium alginate on melanocyte was also investigated. The results show that within the concentration of 64 mmol/L, the cell viability was above 90%, indicating good biocompatibility. Sodium alginate had a certain inhibitory effect on tyrosinase activity and melanin production, which was in a concentration-dependent manner below the concentration of20 mmol/L. The endogenous fluorescence of tyrosinase was partly quenched after interacting with sodium alginate.The redshift of the maximum emission wavelength appearing in the fluorescence spectrum of tyrosinase indicated that the interaction between sodium alginate and tyrosinase changed the hydrophobic environment of its amino acid residues.
引文
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[23]Chai W,Lin M,Song F,et al.Rifampicin as a novel tyrosinase inhibitor:Inhibitory activity and mechanism[J].International Journal of Biological Macromolecules,2017,102:425-430.
[2]Promden W,Viriyabancha W,Monthakantirat O,et al.Correlation between the potency of flavonoids on mushroom tyrosinase inhibitory activity and melanin synthesis in melanocytes[J].Molecules,2018,23(6):1403.
[3]Hwang J,Park N,Na Y,et al.Coumestrol down-regulates melanin production in melan-a murine melanocytes through degradation of tyrosinase[J].Biological&Pharmaceutical Bulletin,2017,40(4):535-539.
[4]Abbas Q,Raza H,Hassan M,et al.Acetazolamide inhibits the level of tyrosinase and melanin:An enzyme kinetic,in vitro,in vivo,and in silico studies[J].Chemistry&Biodiversity,2017,14(9):1700117.
[5]Garcia-Gavin J,Gonzalez-Vilas D,Fernandez-Redondo V,et al.Pigmented contact dermatitis due to kojic acid:A paradoxical side effect of a skin lightener[J].Contact Dermatitis,2010,62(1):63-64.
[6]Mata T,Sanchez J,Oyanguren J.Allergic contact dermatitis due to kojic acid[J].Dermatitis,2005,16(2):89.
[7]Chen C,Lin L,Yang W,et al.An updated organic classification of tyrosinase inhibitors on melanin biosynthesis[J].Current Organic Chemistry,2015,19(1):4-18.
[8]Gasowska-Bajger B,Wojtasek H.Reactions of flavonoids with o-quinones interfere with the spectrophotometric assay of tyrosinase activity[J].Journal of Agricultural and Food Chemistry,2016,64(26):5417-5427.
[9]Lee H,Lee W,Chang S,et al.Hesperidin,a popular antioxidant inhibits melanogenesis via Erk1/2 mediated MITF degradation[J].International Journal of Molecular Sciences,2015,16(8):18384-18395.
[10]Magid A,Voutquenne-Nazabadioko L,Bontemps G,et al.Tyrosinase inhibitors and sesquiterpene diglyclosides from Guioa villosa[J].Planta Medica,2008,74(9):55-60.
[11]Gardelly M,Trimech B,Belkacem M,et al.Synthesis of novel diazaphosphinanes coumarin derivatives with promoted cytotoxic and anti-tyrosinase activities[J].Bioorganic&Medicinal Chemistry Letters,2016,26(10):2450-2454.
[12]Gomes A,Mano J,Queiroz J,et al.Incorporation of antimicrobial peptides on functionalized cotton gauzes for medical applications[J].Carbohydrate Polymers,2015,127:451-461.
[13]Schleeh T,Madau M,Roessner D.Two competing reactions of tetrabutylammonium alginate in organic solvents:Amidation versus gamma-lactone synthesis[J].Carbohydrate Polymers,2016,138:244-251.
[14]Kim H,Song M,Lee E,et al.Injectable hydrogels derived from phosphorylated alginic acid calcium complexes[J].Materials Science and Engineering C,2015,51:139-147.
[15]Vaid U,Mittal S,Babu J.Influence of anion induced proton abstraction on Cu(II)adsorption by alginic acid[J].Reactive and Functional Polymers,2015,97(196):48-55.
[16]Xie L,Chen Q,Huang H,et al.Inhibitory effects of some flavonoids on the activity of mushroom tyrosinase[J].Biochemisty,2003,68(4):481-491.
[17]Aguero L,Zaldivar-Silva D,Pena L,et al.Alginate microparticles as oral colon drug delivery device:A review[J].Carbohydrate Polymer,2017,168:32-24.
[18]Reakasame S,Boccaccini A.Oxidized alginate-based hydrogels for tissue engineering applications:A review[J].Biomacromolecules,2018,19(1):3-21.
[19]Zhou Q,Kang H,Bielec M,et al.Influence of different divalent ions cross-linking sodium alginate-polyacrylamide hydrogels on antibacterial properties and wound healing[J].Carbohydrate Polymers,2018,197:292-304.
[20]Da Costa J,Rodrigues A,Farias L,et al.Biological effects of kojic acid on human monocytes in vitro[J].Biomedicine&Pharmacotherapy,2018,101:100-106.
[21]Higa Y,Kawabe M,Nabae K,et al.Kojic acid-absence of tumorinitiating activity in rat liver,and of carcinogenic and photo-genotoxic potential in mouse skin[J].The Journal of Toxicological Science,2007,32:143-159.
[22]Kus N,Dolinska M,Young K,et al.Membrane-associated human tyrosinase is an enzymatically active monomeric glycoprotein[J].Plos One,2018,13(6):198247.
[23]Chai W,Lin M,Song F,et al.Rifampicin as a novel tyrosinase inhibitor:Inhibitory activity and mechanism[J].International Journal of Biological Macromolecules,2017,102:425-430.