FGF21及其受体FGFR1和FGFR2在小鼠毛囊第1生长周期的表达
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摘要
旨在通过对FGF21、受体FGFR1和FGFR2在小鼠毛囊第1生长周期中的定位和表达情况的研究,以探索FGF21、受体FGFR1和FGFR2在毛囊发育过程中的作用。选取1、3、5、8、12、17、21和23日龄小鼠背部皮肤,采用免疫组织化学技术,实时荧光定量PCR和蛋白免疫印迹技术检测FGF21、FGFR1和FGFR2 mRNA及蛋白表达。结果显示:在小鼠毛囊第1生长周期中,FGF21主要表达于毛囊毛乳头、毛基质、内外根鞘以及毛囊周围的结缔组织中;FGFR1在毛囊各部均有表达,且在退化期(12～17日龄)和静止期(18～21日龄)主要表达于内外根鞘中;FGFR2广泛表达于毛囊各部。1、3、5、8、12、17日龄FGF 21 mRNA表达量极显著低于23日龄(P<0.01),21日龄的表达量低于23日龄,差异不显著(P>0.05);1～5日龄和23日龄FGF21蛋白表达量较高。1、3、5、8、12、21、23日龄FGFR1 mRNA表达量极显著低于17日龄(P<0.01);FGFR1蛋白表达量从12日龄开始上升,在17日龄时达到最高,之后又呈下降趋势。1、5、8、12、21和23日龄FGFR2 mRNA表达量极显著低于17日龄(P<0.01),在3日龄的表达量低于17日龄,差异不显著(P>0.05);FGFR2蛋白在1～17日龄都有较高表达。综上表明,在第1个毛囊生长周期中,FGF21对参与毛囊形成的成纤维细胞的增殖和分化具有重要作用,诱导毛囊由生长期进入退化期。FGFR1对内外根鞘细胞的增殖、分化有着明显作用,诱导毛囊由退化期进入静止期。FGFR2对毛囊内细胞增殖、分化具有重要作用,诱导毛囊由生长期和退化期进入静止期。
To explore the role of FGF21 and receptors FGFR1, FGFR 2 in hair follicle development, we researched the localization and expression of FGF21 and receptors FGFR1, FGFR2 in the first hair follicle growth cycle of mice. The immunohistochemistry, real-time PCR and Western blot were used to study the expression of FGF21 and receptors FGFR1, FGFR2 mRNA and proteins in mice back skin on 1, 3, 5, 8, 12, 17, 21, 23 days old. The results showed that during the first hair follicle growth cycle of mice, FGF21 was mainly expressed in dermal papilla, matrix cells, inner and outer root sheaths and connective tissue around hair follicles; FGFR1 was located in various structures of hair follicle, and abundantly expressed in the inner and outer root sheaths in the catagen(12-17 days old) and telogen(18-21 days old); FGFR2 was abundantly located in various structures of hair follicle. The expression levels of FGF21 mRNA at 1, 3, 5, 8, 12 and 17 days old were significantly lower than at 23 days old(P<0.01), and the expression at 21 days old was lower than at 23 days old, the difference was not significant(P>0.05); The expression levels of FGF21 protein were higher at 1-5 days and 23 days old. The expression levels of FGFR1 mRNA at 1, 3, 5, 8, 12, 21 and 23 days old were significantly lower than at 17 days old(P<0.01); The expression levels of FGFR1 protein increased from 12 days old, reached the highest at 17 days old, and then decreased. The expression levels of FGFR2 mRNA at 1, 5, 8, 12, 21 and 23 days old were significantly lower than at 17 days old(P<0.01), and the expression at 3 days old was lower than at 17 days old, the difference was not significant(P>0.05); FGFR2 protein was highly expressed at 1-17 days old. The results showed that during the first hair follicle growth cycle of mice, FGF21 plays an important role in the proliferation and differentiation of fibroblasts involved in hair follicle formation, and induces hair follicles to enter the catagen from the anagen. FGFR1 plays an important role in the proliferation and differentiation of inner and outer root sheath cells, and induces hair follicles to enter the telogen from the catagen. FGFR2 plays an important role in cell proliferation and differentiation in hair follicles, and induces hair follicles to enter the telogen from the anagen and catagen.
To explore the role of FGF21 and receptors FGFR1, FGFR 2 in hair follicle development, we researched the localization and expression of FGF21 and receptors FGFR1, FGFR2 in the first hair follicle growth cycle of mice. The immunohistochemistry, real-time PCR and Western blot were used to study the expression of FGF21 and receptors FGFR1, FGFR2 mRNA and proteins in mice back skin on 1, 3, 5, 8, 12, 17, 21, 23 days old. The results showed that during the first hair follicle growth cycle of mice, FGF21 was mainly expressed in dermal papilla, matrix cells, inner and outer root sheaths and connective tissue around hair follicles; FGFR1 was located in various structures of hair follicle, and abundantly expressed in the inner and outer root sheaths in the catagen(12-17 days old) and telogen(18-21 days old); FGFR2 was abundantly located in various structures of hair follicle. The expression levels of FGF21 mRNA at 1, 3, 5, 8, 12 and 17 days old were significantly lower than at 23 days old(P<0.01), and the expression at 21 days old was lower than at 23 days old, the difference was not significant(P>0.05); The expression levels of FGF21 protein were higher at 1-5 days and 23 days old. The expression levels of FGFR1 mRNA at 1, 3, 5, 8, 12, 21 and 23 days old were significantly lower than at 17 days old(P<0.01); The expression levels of FGFR1 protein increased from 12 days old, reached the highest at 17 days old, and then decreased. The expression levels of FGFR2 mRNA at 1, 5, 8, 12, 21 and 23 days old were significantly lower than at 17 days old(P<0.01), and the expression at 3 days old was lower than at 17 days old, the difference was not significant(P>0.05); FGFR2 protein was highly expressed at 1-17 days old. The results showed that during the first hair follicle growth cycle of mice, FGF21 plays an important role in the proliferation and differentiation of fibroblasts involved in hair follicle formation, and induces hair follicles to enter the catagen from the anagen. FGFR1 plays an important role in the proliferation and differentiation of inner and outer root sheath cells, and induces hair follicles to enter the telogen from the catagen. FGFR2 plays an important role in cell proliferation and differentiation in hair follicles, and induces hair follicles to enter the telogen from the anagen and catagen.
引文
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[13] CHAPNIK N,GENZER Y,FROY O.Relationship between FGF21 and UCP1 levels under time-restricted feeding and high-fat diet[J].J Nutr Biochem,2017,40:116-121.
[14] HOLLAND W L,ADAMS A C,BROZINICK J T,et al.An FGF21-adiponectin-ceramide axis controls energy expenditure and insulin action in mice[J].Cell Metab,2013,17(5):790-797.
[15] BROOKS A N,KILGOUR E,SMITH P D.Molecular pathways:fibroblast growth factor signaling:a new therapeutic opportunity in cancer[J].Clin Cancer Res,2012,18(7):1855-1862.
[16] GOODMAN L.Getting under your skin[J].J Clin Invest,2004,113(8):1073.
[17] PIRVOLA U,YLIKOSKI J,TROKOVIC R,et al.FGFR1 is required for the development of the auditory sensory epithelium[J].Neuron,2002,35(4):671-680.
[18] TSUBOI R,YAMAZAKI M,MATSUDA Y,et al.Antisense oligonucleotide targeting fibroblast growth factor receptor (FGFR)-1 stimulates cellular activity of hair follicles in an in vitro organ culture system[J].Int J Dermatol,2007,46(3):259-263.
[19] 魏文珍.成纤维细胞生长因子受体2在胃癌中的研究进展[J].医学综述,2016,22(12):2331-2334.WEI W Z.Research progress of FGFR2 and gastric cancer[J].Medical Recapitulate,2016,22(12):2331-2334. (in Chinese)
[20] KELLER U A D,KRAMPERT M,KüMIN A,et al.Keratinocyte growth factor:effects on keratinocytes and mechanisms of action[J].Eur J Cell Biol,2004,83(11-12):607-612.
[21] RICHARDSON G D,BAZZI H,FANTAUZZO K A,et al.KGF and EGF signalling block hair follicle induction and promote interfollicular epidermal fate in developing mouse skin[J].Development,2009,136(13):2153-2164.
[22] MANSUKHANI A,AMBROSETTI D,HOLMES G,et al.Sox2 Induction by FGF and FGFR2 activating mutations inhibits wnt signaling and osteoblast differentiation[J].J Cell Biol,2005,168(7):1065-1076.
[23] PETIOT A,CONTI F J A,GROSE R,et al.A crucial role for FGFR2-IIIb signalling in epidermal development and hair follicle patterning[J].Development,2003,130(22):5493-5501.
[24] ARMAN E,HAFFNER-KRAUSZ R,GORIVODSKY M,et al.Fgfr2 is required for limb outgrowth and lung-branching morphogenesis[J].Proc Natl Acad Sci U S A,1999,96(21):11895-11899.
[25] KHARITONENKOV A,DUNBAR J D,BINA H A,et al.FGF-21/FGF-21 receptor interaction and activation is determined by βKlotho[J].J Cell Physiol,2008,215(1):1-7.
[2] 姜维.太行黑山羊毛囊生长-退行期皮肤组织差异表达基因的筛选与鉴定[D].杨凌:西北农林科技大学,2012.JIANG W.Screening and identifying regulatory genes associated with Taihang black goat hair follicle anagen-catagen transition[D].Yangling:Northwest A&F University,2012. (in Chinese)
[3] BEENKEN A,MOHAMMADI M.The structural biology of the FGF19 subfamily[J].Adv Exp Med Biol,2012,728:1-24.
[4] TOMIYAMA K I,MAEDA R,URAKAWA I,et al.Relevant use of Klotho in FGF19 subfamily signaling system in vivo[J].Proc Natl Acad Sci U S A,2010,107(4):1666-1671.
[5] 张平.利用CRISPR-Cas9技术构建FGF21基因敲除小鼠及其对毛发生长的作用研究[D].呼和浩特:内蒙古大学,2016.ZHANG P.Construction of FGF21 knockout mice using the CRISPR-Cas9 system and the study of its effect of hair growth action[D].Hohhot:Inner Mongolia University,2016. (in Chinese)
[6] 任桂萍,尹杰超,王文飞,等.FGF受体-1和FGF受体-2介导成纤维细胞生长因子-21信号传导[J].中国科学:生命科学,2010,40(4):332-341.REN G P,YIN J C,WANG W F,et al.Fibroblast growth factor (FGF)-21 signals through both FGF receptor-1 and 2[J].Science China: Life Sciences,2010,40(4):332-341.(in Chinese)
[7] 牛姝,程笳琪,高淑媛,等.FGF7亚家族在小鼠毛囊第一生长周期的表达[J].畜牧兽医学报,2018,49(3):515-524.NIU S,CHENG J Q,GAO S Y,et al.Expression profile of fibroblast growth factor 7 sub-family in the first hair follicle cycle of mice skin[J].Acta Veterinaria et Zootechnica Sinica,2018,49(3):515-524. (in Chinese)
[8] 王宁,李进,雷明星,等.c-Myc在小鼠毛囊形成和毛发生长过程中的作用研究[C]//中华医学会医学细胞生物学分会第三届全国学术会议论文集.南宁:中华医学会,2011.WANG N,LI J,LEI M X,et al.The role of c-Myc in mouse hair follicle formation and hair growth[C]//National Academic Conference of Medical Cell Biology Branch of Chinese Medical Association.Nanning:Chinese Medical Association,2011. (in Chinese)
[9] 马馨,陈洋,姜怀志.哺乳动物毛囊结构特征及调控机制[J].中国草食动物科学,2013,33(1):53-56.MA X,CHEN Y,JIANG H Z.Structural characteristics of hair follicles and its regulation mechanism in mammals[J].China Herbivore Science,2013,33(1):53-56. (in Chinese)
[10] 付宏歧.利用转基因烟草和红花表达成纤维细胞生长因子21(FGF21)的研究[D].长春:吉林农业大学,2011.FU H Q.Expression of fibroblast growth factor 21(FGF21) in transgenic tobacco and safflower[D].Changchun:Jilin Agricultural University,2011. (in Chinese)
[11] HONDARES E, IGLESIAS R,GIRALT A,et al.Thermogenic activation induces FGF21 expression and release in brown adipose tissue[J].J Biol Chem,2011,286(15):12983-12990.
[12] JULIAN D,MBBS S,LRCP M.Micrograft size and subsequent survival[J].Dermatol Surg,1997,23(9):757-762.
[13] CHAPNIK N,GENZER Y,FROY O.Relationship between FGF21 and UCP1 levels under time-restricted feeding and high-fat diet[J].J Nutr Biochem,2017,40:116-121.
[14] HOLLAND W L,ADAMS A C,BROZINICK J T,et al.An FGF21-adiponectin-ceramide axis controls energy expenditure and insulin action in mice[J].Cell Metab,2013,17(5):790-797.
[15] BROOKS A N,KILGOUR E,SMITH P D.Molecular pathways:fibroblast growth factor signaling:a new therapeutic opportunity in cancer[J].Clin Cancer Res,2012,18(7):1855-1862.
[16] GOODMAN L.Getting under your skin[J].J Clin Invest,2004,113(8):1073.
[17] PIRVOLA U,YLIKOSKI J,TROKOVIC R,et al.FGFR1 is required for the development of the auditory sensory epithelium[J].Neuron,2002,35(4):671-680.
[18] TSUBOI R,YAMAZAKI M,MATSUDA Y,et al.Antisense oligonucleotide targeting fibroblast growth factor receptor (FGFR)-1 stimulates cellular activity of hair follicles in an in vitro organ culture system[J].Int J Dermatol,2007,46(3):259-263.
[19] 魏文珍.成纤维细胞生长因子受体2在胃癌中的研究进展[J].医学综述,2016,22(12):2331-2334.WEI W Z.Research progress of FGFR2 and gastric cancer[J].Medical Recapitulate,2016,22(12):2331-2334. (in Chinese)
[20] KELLER U A D,KRAMPERT M,KüMIN A,et al.Keratinocyte growth factor:effects on keratinocytes and mechanisms of action[J].Eur J Cell Biol,2004,83(11-12):607-612.
[21] RICHARDSON G D,BAZZI H,FANTAUZZO K A,et al.KGF and EGF signalling block hair follicle induction and promote interfollicular epidermal fate in developing mouse skin[J].Development,2009,136(13):2153-2164.
[22] MANSUKHANI A,AMBROSETTI D,HOLMES G,et al.Sox2 Induction by FGF and FGFR2 activating mutations inhibits wnt signaling and osteoblast differentiation[J].J Cell Biol,2005,168(7):1065-1076.
[23] PETIOT A,CONTI F J A,GROSE R,et al.A crucial role for FGFR2-IIIb signalling in epidermal development and hair follicle patterning[J].Development,2003,130(22):5493-5501.
[24] ARMAN E,HAFFNER-KRAUSZ R,GORIVODSKY M,et al.Fgfr2 is required for limb outgrowth and lung-branching morphogenesis[J].Proc Natl Acad Sci U S A,1999,96(21):11895-11899.
[25] KHARITONENKOV A,DUNBAR J D,BINA H A,et al.FGF-21/FGF-21 receptor interaction and activation is determined by βKlotho[J].J Cell Physiol,2008,215(1):1-7.