牦牛p38MAPK在雌性主要生殖器官中的表达

doi:10.11843/j.issn.0366-6964.2019.09.007

摘要/Abstract

摘要： 旨在研究p38MAPK在母牦牛主要生殖器官中的表达情况，了解其表达差异性，为牦牛的繁殖性能研究提供相关基础资料。本研究采集卵泡期、黄体期、妊娠期成年健康母牦牛（各2头）主要生殖器官样品，免疫组化技术检测p38MAPK蛋白的表达部位；qRT-PCR和Western-blot技术对其基因和蛋白进行相对表达量测定。结果：1）免疫组化结果显示，p38MAPK蛋白在牦牛的卵巢、输卵管和子宫均呈阳性表达；2）qRT-PCR结果表明，p38MAPK基因在卵巢中的表达量卵泡期显著高于妊娠期（P<0.05）；在输卵管中的表达量黄体期显著高于卵泡期和妊娠期（P<0.05）；在子宫中的表达量妊娠期极显著高于卵泡期和黄体期（P<0.01）；3）Western-blot结果显示，p38MAPK蛋白在卵巢中的表达量卵泡期极显著高于黄体期和妊娠期（P<0.01）；在输卵管中的表达量黄体期极显著高于卵泡期和妊娠期（P<0.01）；在子宫中的表达量妊娠期极显著高于卵泡期和黄体期（P<0.01）。结果表明，p38MAPK在母牦牛同一生殖器官的不同繁殖阶段其表达存在差异性，其可能参与了牦牛卵泡发育、黄体功能发挥以及胚胎着床和发育等一系列生殖过程，为牦牛的繁殖性能研究提供相关基础资料。

Abstract: The aim of this study was to explore the expression of p38 mitogen-activated protein kinase (p38MAPK) protein and gene in the main reproductive organs of female yaks, accumulate some fundamental data for comprehending the role of p38MAPK in yak reproductive activity. The samples of main reproductive organs of adult healthy female yak during follicular phase, luteal phase and gestation period(2 each) were collected. Immunohistochemical method was used to locate the expression site of p38MAPK protein in the main reproductive organs of female yaks. The relative expression levels of the p38MAPK gene and protein were detected by qRT-PCR and Western-blot technique, respectively. The results showed that:1) The results of immunohistochemistry showed that p38MAPK protein was positively expressed in ovary, oviduct and uterus in female yak. 2) qRT-PCR results showed that the expression of p38MAPK gene in the ovary was significantly higher during follicular phase than that during gestation period(P<0.05). The expression of p38MAPK gene in oviduct was significantly higher during the luteal phase than that during the follicular phase and gestation period (P<0.05). The expression of p38MAPK gene in the uterus was significantly higher during gestation period than that during follicular phase and luteal phase (P<0.01). 3) The results of Western-blot showed that the expression of p38MAPK protein in the ovary during the follicular phase was significantly higher than that during the luteal phase and gestation period (P<0.01). The expression level of p38MAPK protein in the oviduct during the luteal phase was significantly higher than that during the follicular phase and gestation period (P<0.01). The expression of p38MAPK protein in the uterus during gestation period was significantly higher than that during follicular phase and luteal phase (P<0.01). In conclusion, these results suggest that p38MAPK may be involved in the regulation of reproductive processes that includes follicular development, luteal function and embryo implantation and embryo development. These data provide basic information for researching the reproductive mechanism of female yak.

中图分类号:

S823.85

韩小红, 何翃闳, 王靖雷, 张慧珠, 马悦, 马进彪, 赵生贤, 余四九, 崔燕, 樊江峰. 牦牛p38MAPK在雌性主要生殖器官中的表达[J]. 畜牧兽医学报, 2019, 50(9): 1802-1812.

HAN Xiaohong, HE Honghong, WANG Jinglei, ZHANG Huizhu, MA Yue, MA Jinbiao, ZHAO Shengxian, YU Sijiu, CUI Yan, FAN Jiangfeng. Expression of p38MAPK in the Main Reproductive Organs of Female Yaks (Bos grunniens)[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(9): 1802-1812.

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参考文献

[1] 王亚营,潘阳阳,叶小琳,等.牦牛Cyclin D1基因的克隆及IGF-I对其在睾丸支持细胞表达的影响[J].畜牧兽医学报,2018, 49(2):310-317.
WANG Y Y,PAN Y Y,YE X L,et al.Cloning of yak Cyclin D1 gene and the effects of IGF-I on its expression in sertoli cells[J].Acta Veterinaria et Zootechnica Sinica,2018,49(2):310-317.(in Chinese)
[2] 赵天,李谷月,潘阳阳,等.Oct 4在牦牛卵母细胞及早期胚胎发育过程中的表达[J].畜牧兽医学报,2017,48(3):577-584.
ZHAO T,LI G Y,PAN Y Y,et al.The expression of octamer-binding transcription factor 4 gene (Oct 4) in yak (Bos grunneins) oocytes and preimplantation embryos produced by in vitro fertilization[J].Acta Veterinaria et Zootechnica Sinica,2017, 48(3):577-584.(in Chinese)
[3] 韩杰,熊显荣,蔡雯祎,等.牦牛KDM4A基因克隆、组织表达谱及其在卵母细胞和颗粒细胞中的表达[J].畜牧兽医学报,2018,49(2):291-299.
HAN J,XIONG X R,CAI W Y,et al.Cloning of KDM4A gene and its expression in different tissues,oocyte and granulosa cell of yak[J].Acta Veterinaria et Zootechnica Sinica,2018,49(2):291-299.(in Chinese)
[4] 郭宪,丁学智,王宏博,等.牦牛卵母细胞及体外受精胚胎早期发育基因差异表达的研究[J].畜牧兽医学报,2013,44(10):1576-1581.
GUO X,DING X Z,WANG H B,et al.Gene expression of yak oocytes and preimplantation embryos derived from in vitro fertilization[J].Acta Veterinaria et Zootechnica Sinica,2013,44(10):1576-1581.(in Chinese)
[5] DE ZELICOURT A,COLCOMBET J,HIRT H.The role of MAPK modules and ABA during abiotic stress signaling[J].Trends Plant Sci,2016,21(8):677-685.
[6] 焦今文,赵新卫,邓博雅,等.p38^MAPK对卵巢癌顺铂耐药作用及机制的探讨[J].中华肿瘤防治杂志,2012,19(16):1221-1226.
JIAO J W,ZHAO X W,DENG B Y,et al.Role of the p38^MAPK pathway in cisplatin resistant ovarian cancer and its mechanism[J].Chinese Journal of Cancer Prevention and Treatment,2012,19(16):1221-1226.(in Chinese)
[7] YURTSEVER Z,SCHEAFFER S M,ROMERO A G,et al.The crystal structure of phosphorylated MAPK13 reveals common structural features and differences in p38 MAPK family activation[J].Acta Crystallogr D Biol Crystallogr,2015,71(4):790-799.
[8] 雷杰雯,黄林,金素钰,等.牦牛和雄性不育犏牛睾丸ERK1、ERK2、P38基因mRNA水平的比较[J].四川动物, 2016,35(5):666-671.
LEI J W,HUANG L,JIN S Y,et al.Comparison of the mRNA expression level of ERK1,ERK2 and P38 genes in the testes of yaks and sterile cattle-yaks[J].Sichuan Journal of Zoology,2016,35(5):666-671.(in Chinese)
[9] 郭永娟,郑亚琳,张彩霞,等.MAPK信号通路在动物繁殖中的研究进展[J].中国畜牧兽医,2015,42(5):1268-1273.
GUO Y J,ZHENG Y L,ZHANG C X,et al.Research advance on MAPK signaling pathways in animal reproduction[J].China Animal Husbandry & Veterinary Medicine,2015,42(5):1268-1273.(in Chinese)
[10] 刘晓淋,杜凤娇,雷钏,等.低氧分压通过低氧诱导因子(HIF)影响卵母细胞的体外成熟和早期胚胎的发育[J].中国畜牧兽医, 2015,42(6):1559-1565.
LIU X L,DU F J,LEI C,et al.Effects of low oxygen tension for in vitro oocyte maturation and early embryo development through hypoxia-inducible factor (HIF)[J].China Animal Husbandry & Veterinary Medicine,2015,42(6):1559-1565.(in Chinese)
[11] MINET E,MICHEL G,MOTTET D,et al.Transduction pathways involved in hypoxia-inducible factor-1 phosphorylation and activation[J].Free Radic Biol Med,2001,31(7):847-855.
[12] ALMOG T,NAOR Z.Mitogen activated protein kinases (MAPKs) as regulators of spermatogenesis and spermatozoa functions[J].Mol Cell Endocrinol,2008,282(1-2):39-44.
[13] OHNISHI E,GOTO T,SATO A,et al.Nemo-like kinase,an essential effector of anterior formation,functions downstream of p38 mitogen-activated protein kinase[J].Mol Cell Endocrinol,2010,30(3):675-683.
[14] BREWSTER J L,DE VALORI T,DWYER N D,et al.An osmosensing signal transduction pathway in yeast[J]. Science, 1993,259(5102):1760-1763.
[15] HAN J,LEE J D,BIBBS L,et al.A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells[J].Science,1994,265(5173):808-811.
[16] YANG Y,KIM S C,YU T,et al.Functional roles of p38 mitogen-activated protein kinase in macrophage-mediated inflammatory responses[J].Mediators Inflamm,2014,2014:1-13.
[17] MA K G,LV J,HU X D,et al.The p38 mitogen-activated protein kinase signaling pathway is involved in regulating low-density lipoprotein receptor-related protein 1-mediated β-amyloid protein internalization in mouse brain[J].Int J Biochem Cell Biol, 2016, 76:75-86.
[18] 庄秋宇,刘俊,韩家淮.p38丝裂原活化蛋白激酶的功能与调控机制[J].中国细胞生物学学报,2013,35(2):123-133.
ZHUANG Q Y,LIU J,HAN J H.Functions and mechanisms of the p38 MAP kinase pathway[J].Chinese Journal of Cell Biology, 2013,35(2):123-133.(in Chinese)
[19] 胡世福,夏伟,朱长虹.P38αMAPK在雌性生殖系统中的研究进展[J].国际生殖健康/计划生育杂志,2015,34(1):64-68.
HU S F,XIA W,ZHU C H.P38αMAPK in the female reproductive system[J].Journal of International Reproductive Health/Family Planning,2015,34(1):64-68.(in Chinese)
[20] HU S,YU Q,WANG Y,et al.The localization and function of p38α mitogen-activated protein kinase in rat oocytes[J].Reprod Domest Anim,2018,53(3):636-643.
[21] 李健,陈耀星,陈福宁,等.MAPK信号通路在炔雌醚诱导大鼠生精细胞凋亡中的作用研究[J].畜牧兽医学报,2016,47(2):381-387.
LI J,CHEN Y X,CHEN F N,et al.The role of MAPK signaling pathway in quinestrol-induced apoptosis of spermatogenic cells in rat[J]. Acta Veterinaria et Zootechnica Sinica,2016,47(2):381-387.(in Chinese)
[22] RADI Z A,MARUSAK R A,MORRIS D L.Species comparison of the role of p38 MAP kinase in the female reproductive, system[J].J Toxicol Pathol,2009,22(2):109-124.
[23] 朱宇旌,丁兰,张勇,等.胰岛素对促性腺激素释放激素分泌及活性的调控[J].动物营养学报,2013,25(1):1-7.
ZHU Y J,DING L,ZHANG Y,et al.Regulation of insulin in secretion and activity of gonadotropin-releasing hormone[J].Chinese Journal of Animal Nutrition,2013,25(1):1-7.(in Chinese)
[24] BLISS S P,NAVRATIL A M,XIE J J,et al.GnRH signaling,the gonadotrope and endocrine control of fertility[J].Front Neuroendocrinol, 2010,31(3):322-340.
[25] EWEN K,JACKSON A,WILHELM D,et al.A male-specific role for p38 mitogen-activated protein kinase in germ cell sex differentiation in mice[J].Biol Reprod,2010,83(6):1005-1014.
[26] HU S F,RAO M,LEI H,et al.Expression patterns of p38αMAPK during follicular development in the ovaries of neonatal rats[J]. Acta Histochem,2017,119(5):538-542.
[27] 邹仲之.组织学与胚胎学[M].8版.北京:人民卫生出版社,2013.
ZOU Z Z.Histology and embryology[M].8th ed.Beijing:People's Medical Publishing House,2013.(in Chinese)
[28] 李琦,曹贵方,智达夫,等.LPS对绵羊输卵管上皮细胞β-防御素-1表达的影响及其机制研究[J].中国预防兽医学报,2015, 37(6):477-480.
LI Q,CAO G F,ZHI D F,et al.Study on the beta-defensin 1 expression in ovine oviduct epithelial cells induced by lipopolysaccharide[J]. Chinese Journal of Preventive Veterinary Medicine,2015,37(6):477-480.(in Chinese)
[29] TAKANAMI-OHNISHI Y,ASADA S,TSUNODA H,et al.Possible involvement of p38 mitogen-activated protein kinase in decidual function in parturition[J].Biochem Biophys Res Commun,2001,288(5):1155-1161.
[30] RADI Z A,KHAN N K.Comparative expression and distribution of c-fos,estrogen receptorα (ERα),and p38α in the uterus of rats,monkeys,and humans[J].Toxicol Pathol,2006,34(4):327-335.
[31] KIM J Y.Select nutrients,secreted phosphoprotein 1 and insulin-like growth factor 2:effects of trophectoderm of ovine conceptuses[D].Texas:Texas A&M University,2010.
[32] MADAN P,CALDER M D,WATSON A J.Mitogen-activated protein kinase (MAPK) blockade of bovine preimplantation embryogenesis requires inhibition of both p38 and extracellular signal-regulated kinase (ERK) pathways[J]. Reproduction, 2005, 130(1):41-51.
[33] 于清梅.p38MAPK在小鼠早胚中的表达及其对胚泡发育的作用[J].解剖科学进展,2014,20(2):154-157.
YU Q M.Expression and significance of p38MAPK protein in mouse early embryos[J].Progress of Anatomical Sciences, 2014, 20(2):154-157.(in Chinese)
[34] WANG F W,ZHANG Y M,WANG Z,et al.Calcitonin promotes mouse pre-implantation development:involvement of calcium mobilization and P38 mitogen-activated protein kinase activation[J].Reprod Domest Anim,2013,48(3):382-389.