内皮素-2促进体外培养的绵羊皮肤黑色素细胞增殖及黑色素生成
|
摘要
内皮素(endothelin,ET)及其受体在黑色素细胞成熟分化时起着有效的促进作用.然而,内皮素-2(ET-2)在黑色素生成的作用方面,还处于争论中或报道不一致.在此,我们研究ET-2对体外培养的绵羊皮肤黑色素细胞增殖和黑色素生成的影响.比较实验组ET-2(1,10,100 nmol/L)与空白对照组,通过MTT法和Ando等的方法检测出黑色素细胞的增殖率和黑色素含量显著增加.荧光定量PCR和Western印迹分别检测出内皮素受体B(Bdnrb);酪氨酸激酶受体(Kit);酪氨酸酶(Tyr);酪氨酸相关蛋白-1(Tyrp-1)基因的mRNA水平及蛋白水平的相对表达量显著增加.这些数据表明,ET-2可能促进绵羊的皮肤黑色素细胞增殖和黑色素生成.
Endothelin( ET) and its receptor have been shown to play roles in the melanocyte maturity.However,the reports on the function of ET-2 in melanogenesis are debated. Herein,we investigated the effect of ET-2 on ovine skin melanocyte proliferation and melanogenesis. MTT method and Ando methods showed that the cell proliferation rates and melanin levels were significantly increased in melanocytes treated by ET-2( 1 to 100 nmol / L) compared with the control( untreated) cells. Furthermore,real-time quantitative PCR and Western blotting revealed that the levels of both mRNA and protein of the Bdnrb,Tyr,Kit and Tyrp-1 genes were significantly increased. These data suggest that ET-2 might promote proliferation of ovine skin melanocytes and melanogenesis.
Endothelin( ET) and its receptor have been shown to play roles in the melanocyte maturity.However,the reports on the function of ET-2 in melanogenesis are debated. Herein,we investigated the effect of ET-2 on ovine skin melanocyte proliferation and melanogenesis. MTT method and Ando methods showed that the cell proliferation rates and melanin levels were significantly increased in melanocytes treated by ET-2( 1 to 100 nmol / L) compared with the control( untreated) cells. Furthermore,real-time quantitative PCR and Western blotting revealed that the levels of both mRNA and protein of the Bdnrb,Tyr,Kit and Tyrp-1 genes were significantly increased. These data suggest that ET-2 might promote proliferation of ovine skin melanocytes and melanogenesis.
引文
[1]Yanagisawa M,Kurihara H,Kimura S,et al.A novel potent vasoconstrictor peptide produced by vascular endothelial cell[J].Nature,1988,332(6163):411-415
[2]Slominski A,Tobin D J,Shibahra S,et al.Melanin pigmentaion in mammalian skin and its hormonal regulation[J].Physiol Rev,2004,84:1155-1228
[3]Imokawa G,Yada Y,Miyagishi M.Endothelins secreted from human keratinocytes are intrinsic mitogens for human melanocytes[J].J Biol Chem,1992,267(34):24675–24680
[4]Tsuboi R,Sato C,Oshita Y,et al.Ultraviolet B irradiation increases endothelin-1 and endothelin receptor expression in cultured human keratinocytes[J].FEBS Lett,1995,371(2):188–190
[5]Murakami T,Fujimoto M,Ohtsuki M,et al.Expression profiling of cancer-related genes in human keratinocytes following nonlethal ultraviolet B irradiation[J].J Dermatol Sci,2001,27(2):121–129
[6]Venuti A,Salani D,Poggiali F,et al.The E5 oncoprotein of human papillomavirus type 16 enhances endothelin-1-induced keratinocyte growth[J].Virology,1998,248(1):1–5
[7]Reid K,Turnley AM,Maxwell GD,et al.Multiple roles for endothelin in melanocyte development:regulation of progenitor number and stimulation of differentiation[J].Development,1996,122(12):3911–3919
[8]Kotake-Nara E,Saida K.Endothelin-2/vasoactive intestinal contractor:regulation of expression via reactive oxygen species induced by Co Cl2,and biological activities including neurite outgrowth in PC12 cells[J].Sci World J,2006,6:176–186
[9]Adur J,Takizawa S,Quan J,et al.Increased gene expression and production of murine endothelin receptors after birth[J].Biochem Biophys Res Commun,2003,305(3):700–706
[10]Newton V E.Clinical features of the Waardenburg syndromes[J].Adv Otorhinolaryngol,2002,61:201–208
[11]Bennett DC,Lamoreux ML.The color loci of mice–a genetic century[J].Pigment Cell Res,2003,16(4):333–344
[12]Mosmann T.Rapid colorimetric assay for cellular growth and cytotoxicity assays[J].Immunol Methods,1983,65(1-2):55-63
[13]杨壮群,王正辉,荔鹏,等.芦荟苦素对体外培养的黑素细胞影响的实验研究[J].中国美容医学(Yang Z Q,Wang ZH,Li P,et al.Experimental study of effects of aloesin on cultured human melanocytes in vitro[J].Chin J Aesthetic Med),2003,12(5):464-466
[14]Dupin E,Le Douarin NM.Development of melanocyte precursors from the vertebrate neural crest[J].Oncogene,2003,22(20):3016-3023
[15]Goding CR.Melanocytes:the new Black[J].Int J Biochem Cell Biol,2007,39(2):275-279
[16]Aokih H,Motohashi T,Yoshimura N,et al.Cooperative and indispensable roles of endothelin 3 and KIT signalings in melanocyte[J].Dev Dyn,2005,233(2):407-417
[17]Saldana-Caboverde A,Kos L.Roles of endothelin signaling in melanocyte devlopment and melanoma[J].Pigment Cell Melanona Res,2010,23(2):160-170
[18]耿建军,张杰,穆晓丽,等.内皮素-1(ET-1)对羊驼皮肤黑素细胞增殖和黑素生成的影响[J].畜牧兽医学报(Geng J J,Z J,Mu X L,et al.Effect of ET-1 on proliferation and melanin synthesis of alpaca skin melanocytes in vitro[J].Acta Vet Zootech Sin),2011,42(7):932-938
[19]耿建军,白俊明,范瑞文,等.内皮素-3对羊驼黑色素细胞特征及细胞内毛色基因表达的影响[J].畜牧兽医学报(Geng J J,Bai J M,Fan R W,et al.The effects of endothelin-3 on the melanocyte characteristic and expression of hair color genes in melanocytes from alpaca(Lama Pacos)in vitro[J].Acta Vet Zootech Sin),2013,44(7):1070-1077
[20]Reid K,Turnley A M,Maxwell GD,et al.Multiple roles for endothelin in melanocyte development:regulation of progenitor number and stimulation of differentiation[J].Development,1996,122(2):3911-3919
[2]Slominski A,Tobin D J,Shibahra S,et al.Melanin pigmentaion in mammalian skin and its hormonal regulation[J].Physiol Rev,2004,84:1155-1228
[3]Imokawa G,Yada Y,Miyagishi M.Endothelins secreted from human keratinocytes are intrinsic mitogens for human melanocytes[J].J Biol Chem,1992,267(34):24675–24680
[4]Tsuboi R,Sato C,Oshita Y,et al.Ultraviolet B irradiation increases endothelin-1 and endothelin receptor expression in cultured human keratinocytes[J].FEBS Lett,1995,371(2):188–190
[5]Murakami T,Fujimoto M,Ohtsuki M,et al.Expression profiling of cancer-related genes in human keratinocytes following nonlethal ultraviolet B irradiation[J].J Dermatol Sci,2001,27(2):121–129
[6]Venuti A,Salani D,Poggiali F,et al.The E5 oncoprotein of human papillomavirus type 16 enhances endothelin-1-induced keratinocyte growth[J].Virology,1998,248(1):1–5
[7]Reid K,Turnley AM,Maxwell GD,et al.Multiple roles for endothelin in melanocyte development:regulation of progenitor number and stimulation of differentiation[J].Development,1996,122(12):3911–3919
[8]Kotake-Nara E,Saida K.Endothelin-2/vasoactive intestinal contractor:regulation of expression via reactive oxygen species induced by Co Cl2,and biological activities including neurite outgrowth in PC12 cells[J].Sci World J,2006,6:176–186
[9]Adur J,Takizawa S,Quan J,et al.Increased gene expression and production of murine endothelin receptors after birth[J].Biochem Biophys Res Commun,2003,305(3):700–706
[10]Newton V E.Clinical features of the Waardenburg syndromes[J].Adv Otorhinolaryngol,2002,61:201–208
[11]Bennett DC,Lamoreux ML.The color loci of mice–a genetic century[J].Pigment Cell Res,2003,16(4):333–344
[12]Mosmann T.Rapid colorimetric assay for cellular growth and cytotoxicity assays[J].Immunol Methods,1983,65(1-2):55-63
[13]杨壮群,王正辉,荔鹏,等.芦荟苦素对体外培养的黑素细胞影响的实验研究[J].中国美容医学(Yang Z Q,Wang ZH,Li P,et al.Experimental study of effects of aloesin on cultured human melanocytes in vitro[J].Chin J Aesthetic Med),2003,12(5):464-466
[14]Dupin E,Le Douarin NM.Development of melanocyte precursors from the vertebrate neural crest[J].Oncogene,2003,22(20):3016-3023
[15]Goding CR.Melanocytes:the new Black[J].Int J Biochem Cell Biol,2007,39(2):275-279
[16]Aokih H,Motohashi T,Yoshimura N,et al.Cooperative and indispensable roles of endothelin 3 and KIT signalings in melanocyte[J].Dev Dyn,2005,233(2):407-417
[17]Saldana-Caboverde A,Kos L.Roles of endothelin signaling in melanocyte devlopment and melanoma[J].Pigment Cell Melanona Res,2010,23(2):160-170
[18]耿建军,张杰,穆晓丽,等.内皮素-1(ET-1)对羊驼皮肤黑素细胞增殖和黑素生成的影响[J].畜牧兽医学报(Geng J J,Z J,Mu X L,et al.Effect of ET-1 on proliferation and melanin synthesis of alpaca skin melanocytes in vitro[J].Acta Vet Zootech Sin),2011,42(7):932-938
[19]耿建军,白俊明,范瑞文,等.内皮素-3对羊驼黑色素细胞特征及细胞内毛色基因表达的影响[J].畜牧兽医学报(Geng J J,Bai J M,Fan R W,et al.The effects of endothelin-3 on the melanocyte characteristic and expression of hair color genes in melanocytes from alpaca(Lama Pacos)in vitro[J].Acta Vet Zootech Sin),2013,44(7):1070-1077
[20]Reid K,Turnley A M,Maxwell GD,et al.Multiple roles for endothelin in melanocyte development:regulation of progenitor number and stimulation of differentiation[J].Development,1996,122(2):3911-3919