熊果苷和丙戊酸影响企鹅珍珠贝(Pteria penguin)贝壳色泽
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摘要
企鹅珍珠贝(Pteria penguin)是我国最具潜力培育大型海水珍珠的优质母贝,其贝壳色泽及珍珠层颜色与黑色素(melanin)的合成水平有关。为了筛选安全有效的企鹅珍珠贝黑色素抑制剂和增强剂,探讨其有效使用浓度和作用时间,分析其毒副作用,实验分别采用不同浓度的熊果苷(arbutin)和丙戊酸(valproic acid)浸泡1月龄稚贝不同时间,观察3月龄,5月龄和7月龄幼贝壳色,并统计熊果苷和丙戊酸的使用对幼贝变色率、存活率、壳长、壳高、壳宽的影响。结果显示,2 mmol/L熊果苷处理稚贝1h或5 mmol/L丙戊酸处理1h即可显著提高壳色变异的比率(p<0.05),且幼贝变色率随熊果苷和丙戊酸的浓度和作用时间的增加而增加。对熊果苷和丙戊酸的毒性检测发现,熊果苷达到10 mmol/L处理8h时,仍不会影响幼贝的存活率、壳长、壳高、壳宽(p>0.05);但丙戊酸在浓度为5 mmol/L处理4h时,幼贝的存活率即显著下降,壳长、壳高和壳宽亦显著低于对照(p<0.05)。研究表明,熊果苷和丙戊酸对企鹅珍珠贝幼贝而言分别是有效的黑色素抑制剂和增强剂,熊果苷在10 mmol/L处理8h以下都未检测到毒副作用,丙戊酸在5 mmol/L处理4h时即存在一定毒性。
Pteria penguin is an ideal shellfish to cultivate high-quality seawater pearl,and its shell color and nacre color are related to melanin synthesis.In order to choose an effective and safe medicine to inhibit or enhance melanin level in Pteria penguin,and to study effective concentration,effective action time,and toxicity of the inhibitor and enhancer,the juveniles of 1 month old were immersed into arbutin and valproic acid with 3 different concentration for 1 h,4 h and 8 h.After arbutin and valproic acid treatment,we observed the shell colors of juveniles shellfish with 3 months old,5 months old,and 7 months old,and counted the effects of arbutin and valproic acid on the discoloration rate,survival rate,shell length,shell height and shell width.Research results indicated that treatment with 2 mmol/L arbutin for 1h or 5 mmol/L valproic acid for 1 h could significantly enhance the discoloration rate(p<0.05),which was enhanced with the increased concentration and action time of arbutin and valproic acid.The toxicity detection indicated that arbutin did not affect the survival rate,shell length,shell height and shell width of Pteria penguin juveniles,even if with 10 mmol/L for 8 h(p>0.05).However,the survival rate,shell length,shell height and shell width were significantly decreased when juveniles were disposed with 5 mmol/L valproic acid for 4 h(p<0.05),further more,shell length,shell height and shell width were significantly lower than control(p<0.05).These results suggested that arbutin and valproic acid were effective melanin inhibitor and enhancer for Pteria penguin juveniles,respectively.There was no toxicity detected when arbutin was used with 10 mmol/L arbutin for 8 h or below.However,toxicity was detected when valproic acid was used with5 mmol/L for 4 h.
Pteria penguin is an ideal shellfish to cultivate high-quality seawater pearl,and its shell color and nacre color are related to melanin synthesis.In order to choose an effective and safe medicine to inhibit or enhance melanin level in Pteria penguin,and to study effective concentration,effective action time,and toxicity of the inhibitor and enhancer,the juveniles of 1 month old were immersed into arbutin and valproic acid with 3 different concentration for 1 h,4 h and 8 h.After arbutin and valproic acid treatment,we observed the shell colors of juveniles shellfish with 3 months old,5 months old,and 7 months old,and counted the effects of arbutin and valproic acid on the discoloration rate,survival rate,shell length,shell height and shell width.Research results indicated that treatment with 2 mmol/L arbutin for 1h or 5 mmol/L valproic acid for 1 h could significantly enhance the discoloration rate(p<0.05),which was enhanced with the increased concentration and action time of arbutin and valproic acid.The toxicity detection indicated that arbutin did not affect the survival rate,shell length,shell height and shell width of Pteria penguin juveniles,even if with 10 mmol/L for 8 h(p>0.05).However,the survival rate,shell length,shell height and shell width were significantly decreased when juveniles were disposed with 5 mmol/L valproic acid for 4 h(p<0.05),further more,shell length,shell height and shell width were significantly lower than control(p<0.05).These results suggested that arbutin and valproic acid were effective melanin inhibitor and enhancer for Pteria penguin juveniles,respectively.There was no toxicity detected when arbutin was used with 10 mmol/L arbutin for 8 h or below.However,toxicity was detected when valproic acid was used with5 mmol/L for 4 h.
引文
Cha S.H.,Ko S.C.,Kim D.,and Jeon Y.J.,2011,Screening of marine algae for potential tyrosinase inhibitor:those inhibitors reduced tyrosinase activity and melanin synthesis in zebrafish,J.Dermatol,38(4):354-363
Chodurek E.,Orchel A.,Orchel J.,Kurkiewicz S.,Gawlik N.,Dzierzewicz Z.,and Stepien K.,2012b,Evaluation of melanosenesis in A-375 melanoma cells treated with5,7-dimethoxycoumarin and valproic acid,Cell Mol.Biol.Lett.,17(4):616-632
Chodurek E.,Zdybel M.,Pilawa B.,and Dzierzewicz Z.,2012a,Examination by EPR spectroscopy of free radicals in melanins isolated from A-375 cells exposed on valproic acid and cisplatin,Acta Pol.Pharm.,69(6):1334-1341
Eisinger M.,and Marko O.,1982,Selective poliferation of normal human melanocytes in vitro in the presence of phorbol ester and cholera toxin,Proc.Natl.Acad.Sci.USA,79(6):2018-2022
Kwak S.Y.,Choi H.R.,Park K.C.,and Lee Y.S.,2011,Kojic acid-amino acid amide metal complexes and their melanogenesis inhibitory activities,J.Pept.Sci.,17(12):791-797
Lee J.,Lee J.,Jung E.,Cho J.Y.,and Park D.,2013,Artemisinic acid inhibits melanogenesis through downregulation of C/EBP alpha-dependent expression of HMG-CoA reductase gene,Food Chem.Toxicol.,51:225-230
Lei T.C.,Virador V.,Yasumoto K.,Vieira W.D.,Toyofuku K.,and Hearing V.J.,2002,Stimulation of melanoblast pigmentation by 8-methoxypsoralen:the involvement of microphthalmia-associated transcription factor,the protein kinase a signal pathway,and proteasome-mediated degradation,J.Invest.Dermatol.,119(6):1341-1349
Liao A.H.,Ma W.C.,Wang C.H.,and Yeh M.K.,2014,Penetration depth,concentration and efficiency of transdermalα-arbutin delivery after ultrasound treatment with albumin-shelled microbubbles in mice,Drug Deliv.,21:1-10
Maeda K.,and Fukuda M.,1996,Arbutin:mechanism of its depigmenting action in human melanocyte culture,J.Pharmacol.Exp.Ther.,276(2):765-769
Masse M.O.,Duvallet V.,Borremans M.,and Goeyens L.,2001,Identification and quantitative analysis of kojic acid and arbutine in skin-whitening cosmetics,Int.J.Cosmet.Sci.,23(4):219-232
Sasaki C,Ichitani M.,Kunimoto K.K.,Asada C,and Nakamura Y.,2014,Extraction of arbutin and its comparative content in branches,leaves,stems,and fruits of Japanese pear Pyrus pyrifolia cv.Kousui,Biosci.Biotechnol.Biochem.,78(5):874-877
Sugimoto K.,Nishimura T.,Nomure K.,Sugimoto K.,and Kuriki T.,2004,Inhibitory effectis of a-arbutin on melamin synthesis in cultured human melanoma cells and a three-dimensional human skin model,Biol.Pharm.Bull,27(4):510-514
Takgi R.,and Miyashita T.,2014,A cDNA cloning of novel alpha-class tyrosinase of Pinctada fucata:its expression analysis and characterization of the expressed procin,Enzyme Res.,2014:780549
Tryon R.C.,and Johnson S.L.,2012,Clonal and lineage analysis of melanocyte stem cells and their progeny in the zebrafish,Methods Mot.Biol.,916:181-195
Wang M.F.,Yu F.Y.,Wang Y.L.Wang A.M.,and Yu X.Y.,2015,The microsatellte analysis and shell traits analysis in Pteria penguin from cultivated population and wild population of weizhou island,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(9):1887-1895(王梅芳,于非非,王永丽,王爱民,余祥勇,2015,企鹅珍珠贝养殖群体与涸洲岛野生群体的微卫星分析和形态分析。基因组学与应用生物学,34(9):1887-1895)
Wang Y.L.,2010,Studies on microsatellite marker and morphological contrastive analysis of wild and cultivated Pteria penguin(Boding),Thesis for M:S.,Guangdong Ocean University,Suppervisor:Wang M.F.,pp.25-26(王永丽,2010,企鹅珍珠贝野生与养殖群体的微卫星标记及形态学对比分析,硕士学位论文,广东海洋大学,导师:王梅芳,PP25-26)
Xu M.,Lao Q.C.,Zhao P.,Zhu X.Y.;Zhu H.T.,Luo X.L.,Yang C.R.,He J.H.,Li C.Q.,and Zhang Y.T.,2014,6'-O-Caffeoylarbutin inhibits melanogenesis in zebradsh,Nat.Prod.Res.,28(12):932-934
Yao C.,Oh J.H.,Oh I.G.,Park C.M.,and Chung J.H.,2013,[6]-Shogaol inhibits melanogenesis in B16 mouse melanoma cells through activation of the ERK pathway,Acta Pharmacol.Sin.,34(2):289-294
Yoon T.T,and Hearing V.T.,2003,Co-culture of mouse epidermal cells for studies of pigmentation,Pigment Cell Res.,16(2):159-163
Yu X.,Yu H.,Kong L.,Guo F.,Zhu G.,and Li Q.,2014,Molecular cloning and differential expression in tissues of a tyrosi ease gene in Pacific oyster Crassostrea ggas,Mol.Biol:Rep.,41(8):5403-5411
Yu X.Y.,Wang M.F.,Liang F.L.,and Liu Y.,2005,Differentiation of isozymes from different tissues of Pteria penguin coding,Zhongguo Shuichan Kexue(Journal of Fishery Sciences of China),12(2):201-206(余祥勇,王梅芳:梁飞龙:刘永,2005,企鹅珍珠贝不同组织同工酶表达的差异,中国水产科学,12(2):201-206)
Zdybel M.,Chodurek E.,and Pilawa B.,2014,EPR studies of free radicals in A-2058 human melanoma cells treated by valpromic acid and 5,7-dimethoxycoumarin,Acta Pol.Pharm.,71(6):1066-1072
Chodurek E.,Orchel A.,Orchel J.,Kurkiewicz S.,Gawlik N.,Dzierzewicz Z.,and Stepien K.,2012b,Evaluation of melanosenesis in A-375 melanoma cells treated with5,7-dimethoxycoumarin and valproic acid,Cell Mol.Biol.Lett.,17(4):616-632
Chodurek E.,Zdybel M.,Pilawa B.,and Dzierzewicz Z.,2012a,Examination by EPR spectroscopy of free radicals in melanins isolated from A-375 cells exposed on valproic acid and cisplatin,Acta Pol.Pharm.,69(6):1334-1341
Eisinger M.,and Marko O.,1982,Selective poliferation of normal human melanocytes in vitro in the presence of phorbol ester and cholera toxin,Proc.Natl.Acad.Sci.USA,79(6):2018-2022
Kwak S.Y.,Choi H.R.,Park K.C.,and Lee Y.S.,2011,Kojic acid-amino acid amide metal complexes and their melanogenesis inhibitory activities,J.Pept.Sci.,17(12):791-797
Lee J.,Lee J.,Jung E.,Cho J.Y.,and Park D.,2013,Artemisinic acid inhibits melanogenesis through downregulation of C/EBP alpha-dependent expression of HMG-CoA reductase gene,Food Chem.Toxicol.,51:225-230
Lei T.C.,Virador V.,Yasumoto K.,Vieira W.D.,Toyofuku K.,and Hearing V.J.,2002,Stimulation of melanoblast pigmentation by 8-methoxypsoralen:the involvement of microphthalmia-associated transcription factor,the protein kinase a signal pathway,and proteasome-mediated degradation,J.Invest.Dermatol.,119(6):1341-1349
Liao A.H.,Ma W.C.,Wang C.H.,and Yeh M.K.,2014,Penetration depth,concentration and efficiency of transdermalα-arbutin delivery after ultrasound treatment with albumin-shelled microbubbles in mice,Drug Deliv.,21:1-10
Maeda K.,and Fukuda M.,1996,Arbutin:mechanism of its depigmenting action in human melanocyte culture,J.Pharmacol.Exp.Ther.,276(2):765-769
Masse M.O.,Duvallet V.,Borremans M.,and Goeyens L.,2001,Identification and quantitative analysis of kojic acid and arbutine in skin-whitening cosmetics,Int.J.Cosmet.Sci.,23(4):219-232
Sasaki C,Ichitani M.,Kunimoto K.K.,Asada C,and Nakamura Y.,2014,Extraction of arbutin and its comparative content in branches,leaves,stems,and fruits of Japanese pear Pyrus pyrifolia cv.Kousui,Biosci.Biotechnol.Biochem.,78(5):874-877
Sugimoto K.,Nishimura T.,Nomure K.,Sugimoto K.,and Kuriki T.,2004,Inhibitory effectis of a-arbutin on melamin synthesis in cultured human melanoma cells and a three-dimensional human skin model,Biol.Pharm.Bull,27(4):510-514
Takgi R.,and Miyashita T.,2014,A cDNA cloning of novel alpha-class tyrosinase of Pinctada fucata:its expression analysis and characterization of the expressed procin,Enzyme Res.,2014:780549
Tryon R.C.,and Johnson S.L.,2012,Clonal and lineage analysis of melanocyte stem cells and their progeny in the zebrafish,Methods Mot.Biol.,916:181-195
Wang M.F.,Yu F.Y.,Wang Y.L.Wang A.M.,and Yu X.Y.,2015,The microsatellte analysis and shell traits analysis in Pteria penguin from cultivated population and wild population of weizhou island,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(9):1887-1895(王梅芳,于非非,王永丽,王爱民,余祥勇,2015,企鹅珍珠贝养殖群体与涸洲岛野生群体的微卫星分析和形态分析。基因组学与应用生物学,34(9):1887-1895)
Wang Y.L.,2010,Studies on microsatellite marker and morphological contrastive analysis of wild and cultivated Pteria penguin(Boding),Thesis for M:S.,Guangdong Ocean University,Suppervisor:Wang M.F.,pp.25-26(王永丽,2010,企鹅珍珠贝野生与养殖群体的微卫星标记及形态学对比分析,硕士学位论文,广东海洋大学,导师:王梅芳,PP25-26)
Xu M.,Lao Q.C.,Zhao P.,Zhu X.Y.;Zhu H.T.,Luo X.L.,Yang C.R.,He J.H.,Li C.Q.,and Zhang Y.T.,2014,6'-O-Caffeoylarbutin inhibits melanogenesis in zebradsh,Nat.Prod.Res.,28(12):932-934
Yao C.,Oh J.H.,Oh I.G.,Park C.M.,and Chung J.H.,2013,[6]-Shogaol inhibits melanogenesis in B16 mouse melanoma cells through activation of the ERK pathway,Acta Pharmacol.Sin.,34(2):289-294
Yoon T.T,and Hearing V.T.,2003,Co-culture of mouse epidermal cells for studies of pigmentation,Pigment Cell Res.,16(2):159-163
Yu X.,Yu H.,Kong L.,Guo F.,Zhu G.,and Li Q.,2014,Molecular cloning and differential expression in tissues of a tyrosi ease gene in Pacific oyster Crassostrea ggas,Mol.Biol:Rep.,41(8):5403-5411
Yu X.Y.,Wang M.F.,Liang F.L.,and Liu Y.,2005,Differentiation of isozymes from different tissues of Pteria penguin coding,Zhongguo Shuichan Kexue(Journal of Fishery Sciences of China),12(2):201-206(余祥勇,王梅芳:梁飞龙:刘永,2005,企鹅珍珠贝不同组织同工酶表达的差异,中国水产科学,12(2):201-206)
Zdybel M.,Chodurek E.,and Pilawa B.,2014,EPR studies of free radicals in A-2058 human melanoma cells treated by valpromic acid and 5,7-dimethoxycoumarin,Acta Pol.Pharm.,71(6):1066-1072