雌激素受体在泌乳中期和静止期牦牛乳腺组织内的分布与表达
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摘要
旨在揭示牦牛泌乳中期和静止期乳腺组织雌激素受体α(ERα)和雌激素受体β(ERβ)的分布及蛋白水平的表达。采集泌乳中期(分娩后3～4个月)和静止期(断乳后1个月)各5头健康牦牛的乳腺组织,应用SP免疫组织化学染色方法和Image pro plus 6.0图像分析软件检测ERα和ERβ在牦牛泌乳中期和静止期的分布情况,并采用Western blot技术对乳腺组织内的ERα和ERβ蛋白含量进行半定量测定。结果表明:泌乳中期牦牛乳腺组织内ERα大多分布于腺泡上皮细胞的细胞质、少部分分布于小叶间结缔组织内脂肪细胞的细胞核、偶见于血管内皮细胞的细胞核;静止期牦牛乳腺组织内ERα大量分布于结缔组织中脂肪细胞及血管内皮细胞的细胞核中,少数分布于腺泡上皮细胞的细胞质中;泌乳中期ERβ大多分布于乳腺腺泡上皮细胞的细胞质,偶见于血管内皮细胞及脂肪细胞的细胞核中,但着色较腺泡上皮细胞的细胞质浅。静止期ERβ广泛分布于乳腺导管上皮细胞的细胞质、小叶间结缔组织内间质细胞的细胞核及血管内皮细胞的细胞核中,腺泡上皮细胞的细胞质内也有表达,但较泌乳中期阳性细胞数少。牦牛乳腺组织内ERα蛋白的相对表达量静止期显著高于泌乳中期(P<0.05);静止期与泌乳中期乳腺组织内ERβ蛋白的相对表达量无显著差异(P>0.05)。雌激素受体不同亚型在不同发育期牦牛乳腺组织内的表达存在差异,提示,雌激素受体不同亚型在牦牛乳腺发育及泌乳生物学中可能执行不同的功能。
In order to reveal the distribution and expression of estrogen receptorα (ERα) and estrogen receptorβ (ERβ) in mammary gland of yak during mid-lactating and non-lactating periods.The mammary tissues from each of 5healthy yaks in the mid-lactating (3-4months after parturition) and non-lactating periods (1month after weaning) were collected,the localization,distribution and protein content of ERαand ERβwere tested by using the SP immunohistochemical staining method and the Western blot method,respectively.The results showed that predominantcytoplasmic staining for ERαwere found in gland bubble epithelial cells and weak nuclear staining for ERαwere found in fat cells in interlobular connective tissue and vascular endothelial cells in mammary gland during mid-lactating period.But during non-lactating period,distribution of ERα-positive material was contrary to that in mid-lactating period.Predominant cytoplasmic staining for ERβwere found in the gland bubble epithelial cells during mid-lactating period and weak nucleus staining for ERβwere found in vascular endothelial cells and fat cells.During non-lactating period,ERβ-positive material were found in cytoplasm of duct epithelial cells,in the nucleus of stroma cells in interlobular connective tissue and vascular endothelial cells,and in cytoplasm of acinar epithelial cells,but the number of positive cells was less than that during mid-lactating period.The relative expression of ERαprotein in yak mammary gland tissue in non-lactating period was significantly higher than that in mid-lactating period (P<0.05),while there was no significant difference in the relative expression of ERβprotein between mid-lactating and non-lactating periods (P>0.05).The different expression patterns of estrogen receptor subtypes in mammary gland of yak in different periods suggested that ERαand ERβin mammary gland of yak might perform different functions during the process of mammary development and lactation.
In order to reveal the distribution and expression of estrogen receptorα (ERα) and estrogen receptorβ (ERβ) in mammary gland of yak during mid-lactating and non-lactating periods.The mammary tissues from each of 5healthy yaks in the mid-lactating (3-4months after parturition) and non-lactating periods (1month after weaning) were collected,the localization,distribution and protein content of ERαand ERβwere tested by using the SP immunohistochemical staining method and the Western blot method,respectively.The results showed that predominantcytoplasmic staining for ERαwere found in gland bubble epithelial cells and weak nuclear staining for ERαwere found in fat cells in interlobular connective tissue and vascular endothelial cells in mammary gland during mid-lactating period.But during non-lactating period,distribution of ERα-positive material was contrary to that in mid-lactating period.Predominant cytoplasmic staining for ERβwere found in the gland bubble epithelial cells during mid-lactating period and weak nucleus staining for ERβwere found in vascular endothelial cells and fat cells.During non-lactating period,ERβ-positive material were found in cytoplasm of duct epithelial cells,in the nucleus of stroma cells in interlobular connective tissue and vascular endothelial cells,and in cytoplasm of acinar epithelial cells,but the number of positive cells was less than that during mid-lactating period.The relative expression of ERαprotein in yak mammary gland tissue in non-lactating period was significantly higher than that in mid-lactating period (P<0.05),while there was no significant difference in the relative expression of ERβprotein between mid-lactating and non-lactating periods (P>0.05).The different expression patterns of estrogen receptor subtypes in mammary gland of yak in different periods suggested that ERαand ERβin mammary gland of yak might perform different functions during the process of mammary development and lactation.
引文
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[19]李嗣杰,韩冰,范志民,等.雌激素受体α和β在人正常乳腺组织中的表达及意义[J].吉林大学学报:医学版,2009,35(2):356-359.LI S J,HAN B,FAN Z M,et al.Expression of ERαand ERβin normal human mammary gland and significance[J].Journal of Jilin University:Medicine Edition,2009,35(2):356-359.(in Chinese)
[20]ZEPS N,BENTEL J M,PAPADIMITRIOU J M,et al.Murine progesterone receptor expression in proliferating mammary epithelial cells during normal pubertal development and adult estrous cycle.Association with ERαand ERβstatus[J].J Histochem Cytochem,1999,47(10):1323-1330.
[21]KREGE J H,HODGIN J B,COUSE J F,et al.Generation and reproductive phenotypes of mice lacking estrogen receptorβ[J].Proc Natl Acad Sci U S A,1998,95(26):15677-15682.
[22]SAJI S,SAKAGUCHI H,ANDERSSON S,et al.Quantitative analysis of estrogen receptor proteins in rat mammary gland[J].Endocrinology,2001,142(7):3177-3186.
[2]黄筱声.高原美味雪域珍品-牦牛食品前景广阔[J].中国乳品工业,2000,28(6):14-16.HUANG X S.The delicacy and treasure from the plateau bos Grunniens with its broad prospect[J].China Dairy Industry,2000,28(6):14-16.(in Chinese)
[3]郭宪,裴杰,王宏博,等.牦牛乳及乳产品的研究与开发利用[J].安徽农业科学,2014,42(19):6256-6257.GUO X,PEI J,WANG H B,et al.Research and development of yak milk and dairy products[J].Journal of Anhui Agricultural Sciences,2014,42(19):6256-6257.(in Chinese)
[4]NAYLOR M J,OAKES S R,GARDINER-GARDENM,et al.Transcriptional changes underlying the secretory activation phase of mammary gland development[J].Mol Endocrinol,2005,19(7):1868-1883.
[5]SHYAMALA G,CHOU Y C,LOUIE S G,et al.Cellular expression of estrogen and progesterone receptors in mammary glands:regulation by hormones,development and aging[J].J Steroid Biochem Mol Biol,2002,80(2):137-148.
[6]赵晓民,徐小明.雌激素受体及其作用机制[J].西北农林科技大学学报:自然科学版,2004,32(12):154-158.ZHAO X M,XU X M.Estrogen receptor and its molecular mechanism[J].Journal of Northwest A&FUniversity:Natural Science Edition,2004,32(12):154-158.(in Chinese)
[7]尹玉涛,王树迎,刘宵,等.出生后发育时期济宁青山羊乳腺中ERα和PR的分布及其mRNA表达[J].畜牧兽医学报,2012,43(12):1984-1990.YIN Y T,WANG S Y,LIU X,et al.Localization of ERαand PR and differential expression of ERαmRNA and PR mRNA in mammary gland of Jining gray goat during postnatal development[J].Acta Veterinaria et Zootechnica Sinica,2012,43(12):1984-1990.(in Chinese)
[8]李键,符梅,兰道亮,等.牦牛雌激素受体ERα和ERβ基因分子特征及组织表达研究[J].西南民族大学学报:自然科学版,2014,40(6):801-808.LI J,FU M,LAN D L,et al.Molecular characterization and tissue distribution of ERαand ERβgenes in domestic yak[J].Journal of Southwest University for Nationalities:Natural Science Edition,2014,40(6):801-808.(in Chinese)
[9]COUSE J F,LINDZEY J,GRANDIEN K,et al.Tissue distribution and quantitative analysis of estrogen receptor-α(ERα)and estrogen receptor-β(ERβ)messenger ribonucleic acid in the wild-type and ERα-knockout mouse[J].Endocrinology,1997,138(11):4613-4621.
[10]BOWERS J L,TYULMENKOV V V,JERNIGAN SC,et al.Resveratrol acts as a mixed agonist/antagonist for estrogen receptorsαandβ[J].Endocrinology,2000,141(10):3657-3667.
[11]KUIPER G G J M,CARLSSON B,GRANDIEN K,et al.Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptorsαandβ[J].Endocrinology,1997,138(3):863-870.
[12]SAJI S,JENSENE E V,NILSSON S,et al.Estrogen receptorsαandβin the rodent mammary gland[J].Proc Natl Acad Sci U S A,2000,97(1):337-342.
[13]SCHAMS D,KOHLENBERG S,AMSELGRUBERW,et al.Expression and localisation of oestrogen and progesterone receptors in the bovine mammary gland during development,function and involution[J].JEndocrinol,2003,177(2):305-317.
[14]徐鹰妮,姜军,程鸿,等.ERα和ERβ在不同类型乳腺组织的表达及其意义[J].第三军医大学学报,2007,29(11):1093-1095.XU Y N,JIANG J,CHENG H,et al.Expression and significance of estrogen receptor isoforms in different breast tissues[J].Acta Academiae Medicinae Militaris Tertiae,2007,29(11):1093-1095.(in Chinese)
[15]FENG Y X,MANKA D,WAGNER K U,et al.Estrogen receptor-αexpression in the mammary epithelium is required for ductal and alveolar morphogenesis in mice[J].Proc Natl Acad Sci U S A,2007,104(37):14718-14723.
[16]李楠,高学军.雌激素及其受体对动物乳腺上皮细胞泌乳性能的影响[J].中国畜牧兽医,2011,38(4):151-153.LI N,GAO X J.Effects of estrogen and its receptor on lactation fuction of mammary gland epithelial cell in mammal[J].China Animal Husbandry&Veterinary Medicine,2011,38(4):151-153.(in Chinese)
[17]杜娟.奶牛乳腺上皮细胞高温应答机制的研究[D].南京:南京农业大学,2007.DU J.Mechanism of bovine mammary epithelial cells response to hyperthermia[D].Nanjing:Nanjing Agricultural University,2007.(in Chinese)
[18]谈智,王庭槐.雌激素作用分子机制研究进展[J].中国病理生理杂志,2003,19(10):1422-1426.TAN Z,WANG T H.Mechanisms of estrogen action[J].Chinese Journal of Pathophysiology,2003,19(10):1422-1426.(in Chinese)
[19]李嗣杰,韩冰,范志民,等.雌激素受体α和β在人正常乳腺组织中的表达及意义[J].吉林大学学报:医学版,2009,35(2):356-359.LI S J,HAN B,FAN Z M,et al.Expression of ERαand ERβin normal human mammary gland and significance[J].Journal of Jilin University:Medicine Edition,2009,35(2):356-359.(in Chinese)
[20]ZEPS N,BENTEL J M,PAPADIMITRIOU J M,et al.Murine progesterone receptor expression in proliferating mammary epithelial cells during normal pubertal development and adult estrous cycle.Association with ERαand ERβstatus[J].J Histochem Cytochem,1999,47(10):1323-1330.
[21]KREGE J H,HODGIN J B,COUSE J F,et al.Generation and reproductive phenotypes of mice lacking estrogen receptorβ[J].Proc Natl Acad Sci U S A,1998,95(26):15677-15682.
[22]SAJI S,SAKAGUCHI H,ANDERSSON S,et al.Quantitative analysis of estrogen receptor proteins in rat mammary gland[J].Endocrinology,2001,142(7):3177-3186.