母牦牛生殖系统中低氧诱导因子基因的表达分析
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摘要
低氧诱导因子1α(Hypoxia-inducible factor 1α,HIF-1α)和2α(Hypoxia-inducible factor 2α,HIF-2α)是诱导低氧基因和修复细胞氧内环境的核心调节因子,为了研究生活在青藏高原牦牛的繁殖机能对低氧环境的适应机制,采集卵泡期的5头成年母牦牛和母黄牛的下丘脑、脑垂体、卵巢、输卵管和子宫组织,利用RT-qPCR技术检测HIF-1α和HIF-2αm RNA的表达量。结果表明:HIF-1αmRNA在牦牛输卵管中表达量最高,牦牛脑垂体和输卵管表达量极显著高于黄牛(P<0.01)。HIF-2αmRNA在牦牛脑垂体表达量极显著高于黄牛(P<0.01),在输卵管表达量显著高于黄牛(P<0.05)。说明在低氧环境条件下,母牦牛可能通过调节生殖生殖系统中HIF-1α和HIF-2α基因的表达维持其繁殖机能。
Hypoxia-inducible factor 1α(HIF-1α)and 2α(HIF-2α)are core regulators for inducing hypoxia genes and repairing cellular oxygen environment. This study aims to investigate the adaptation mechanism of the reproduction of yak living on the Qinghai-Tibet Plateau to the hypoxic environment. Hypothalamus,pituitary,oviduct,ovary and uterus tissue samples were collected from five adult and health female yaks and cattle during the follicular phase,respectively. Tissue expression characteristics of these genes were performed by RT-qPCR. The results showed that the mRNA expression level of HIF-1α in the oviduct was the highest,and was very significantly higher in the pituitary and oviduct in yak than in cattle(P<0.01). Meanwhile,the mRNA expression of HIF-2α was very significantly higher in the hypothalamus(P<0.01)and significantly higher in the ovary of yak than in cattle(P<0.05). These results suggest that female yaks maintain their reproductive function mainly by regulating HIF-1α and HIF-2α genes in the reproductive system under hypoxic conditions.
Hypoxia-inducible factor 1α(HIF-1α)and 2α(HIF-2α)are core regulators for inducing hypoxia genes and repairing cellular oxygen environment. This study aims to investigate the adaptation mechanism of the reproduction of yak living on the Qinghai-Tibet Plateau to the hypoxic environment. Hypothalamus,pituitary,oviduct,ovary and uterus tissue samples were collected from five adult and health female yaks and cattle during the follicular phase,respectively. Tissue expression characteristics of these genes were performed by RT-qPCR. The results showed that the mRNA expression level of HIF-1α in the oviduct was the highest,and was very significantly higher in the pituitary and oviduct in yak than in cattle(P<0.01). Meanwhile,the mRNA expression of HIF-2α was very significantly higher in the hypothalamus(P<0.01)and significantly higher in the ovary of yak than in cattle(P<0.05). These results suggest that female yaks maintain their reproductive function mainly by regulating HIF-1α and HIF-2α genes in the reproductive system under hypoxic conditions.
引文
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[25]Agarwal A,Gupta S,Sekhon L,et al.Redox considerations in female reproductive function and assisted reproduction:from molecular mechanisms to health implications[J].Antioxidants&Redox Signaling,2008,10(8):1375-1404.
[26]Mirzaei Bavil F,Alipour MR,Keyhanmanesh R,et al.Ghrelin decreases angiogenesis,HIF-1αand VEGF protein levels in chronic hypoxia in lung tissue of male rats[J].Advanced Pharmaceutical Bulletin,2015,5(3):315-320.
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[28]Li Y,Padmanabha D,Gentile LB,et al.HIF-and non-HIF-regulated hypoxic responses require the estrogen-related receptor in Drosophila melanogaster[J].PLoS Genetics,2013,9(1):e1003230.
[2]杨梦思,周娜,王志钢,等.转录因子HIF-1α及其信号通路在疾病发生中的作用研究进展[J].生物技术通报,2016,32(8):8-13.
[3]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.
[4]陈香梅,熊艳蕾,龚辉,等.低氧下HIF-1α对小鼠性激素及睾丸生精细胞凋亡的影响[J].中国应用生理学杂志,2013,29(4):371-375.
[5]Favier J,Kempf H,Corvol P,et al.Cloning and expression pattern of EPAS1 in the chicken embryo:Colocalization with Tyrosine Hydroxylase[J].Febs Letters,2018,462(1-2):19-24.
[6]Matsumoto L,Hirota Y,Saito-Fujita T,et al.HIF2αin the uterine stroma permits embryo invasion and luminal epithelium detachment[J].Journal of Clinical Investigation,2018,128(7):3186-3197.
[7]魏莲,王辰龙,刘力华,等.低氧对不同海拔雄性高原鼠兔生殖机能若干指标的影响[J].青海大学学报:自然科学版,2014,32(3):1-5.
[8]Lan D,Xiong X,Huang C,et al.Toward understanding the genetic basis of yak ovary reproduction:a characterization and comparative analyses of estrus ovary transcriptome in yak and cattle[J].PLoSOne,2016,11(4):e0152675.
[9]Gerald W,Han JL,Long RJ.The yak[J].RAP Publication:Thailand,2003.
[10]Zhou GL,Jin HG,Zhu Q,et al.Genetic diversity analysis of five cattle breeds native to China using microsatellites[J].Journal of Genetics,2005,84(1):77-80.
[11]王德朋,李红阁,郭松长,等.青海家牦牛HIF-1α基因组织特异性表达[J].安徽农业科学,2007,35(29):9173-9175.
[12]Bionaz M,Loor JJ.Gene networks driving bovine milk fat synthesis during the lactation cycle[J].BMC Genomics,2008,9(1):366-10.1186/1471-2164-9-366.
[13]姜华鹏,张驰,王丛丛,等.软刺裸鲤和齐口裂鳆鱼HIF1B和HIF2A的克隆及低氧适应性的表达分析[J].淡水渔业,2015,45(5):11-18.
[14]周丽娟.缺氧对人滋养细胞中MAP4及微管的影响[C].中华高血压杂志社.中国转化医学和整合医学研讨会论文综合刊,2015.
[15]Matsubara K.Hypoxia in the pathogenesis of preeclampsia[J].Hypertension Research in Pregnancy,2017,5(2):46-51.
[16]Murray AJ.Oxygen delivery and fetal-placental growth:beyond a question of supply and demand[J].Placenta,2012,33(2):16-22.
[17]Bangham CR,Sacherer JM.Fertility of Nepalese Sherpas at moderate altitudes:comparison with high-altitude data[J].Annals of Human Biology,1980,7(4):323-330.
[18]Jiang YF,Tsui KH,Wang PH,et al.Hypoxia regulates cell proliferation and steroidogenesis through protein kinase A signaling in bovine corpus luteum[J].Animal Reproduction Science,2011,129(3-4):152-161.
[19]Parraguez VH,Atlagich MA,Behn C,et al.Fertility in ewes at high altitude:comparison between animals with long-and shorttime residence at high altitude and the effect of antioxidant vitamins[J].Reproduction in Domestic Animals,2006,41(4):372-372.
[20]Zhao TB,Ning H X,Zhu SS,et al.Cloning of hypoxia-inducible factor 1αcDNA from a high hypoxia tolerant mammal-plateau pika(Ochotona curzoniae)[J].Biochemical and Biophysical Research Communications,2004,316(2):565-572.
[21]Xiong X,Fu M,Lan D,et al.Yak response to high-altitude hypoxic stress by altering mRNA expression and DNA methylation of hypoxia-inducible factors[J].Animal Biotechnology,2015,26(3):222-229.
[22]Hunzicker-Dunn M,Maizels ET.FSH signaling pathways in immature granulosa cells that regulate target gene expression:branching out from protein kinase A[J].Cellular Signalling,2006,18(9):1351-1359.
[23]Lee HG,Lee HC,Chung HJ,et al.Study on the reproductive function in transgenic pig Harboring Human Erythropoietin(HEPO)gene[J].Reproductive&Developmental Biology,2008,32(2):117-121.
[24]Ma Y,Freitag P,Zhou J,et al.Thyroid hormone induces erythropoietin gene expression through augmented accumulation of hypoxia-inducible factor-1[J].American Journal of PhysiologyRegulatory,Integrative and Comparative Physiology,2004,287(3):R600-R607.
[25]Agarwal A,Gupta S,Sekhon L,et al.Redox considerations in female reproductive function and assisted reproduction:from molecular mechanisms to health implications[J].Antioxidants&Redox Signaling,2008,10(8):1375-1404.
[26]Mirzaei Bavil F,Alipour MR,Keyhanmanesh R,et al.Ghrelin decreases angiogenesis,HIF-1αand VEGF protein levels in chronic hypoxia in lung tissue of male rats[J].Advanced Pharmaceutical Bulletin,2015,5(3):315-320.
[27]Hüppi PS.The role of oxygen in health and disease-a series of reviews[J].Pediatric Research,2009,65(3):261-268.
[28]Li Y,Padmanabha D,Gentile LB,et al.HIF-and non-HIF-regulated hypoxic responses require the estrogen-related receptor in Drosophila melanogaster[J].PLoS Genetics,2013,9(1):e1003230.