缺氧诱导因子和促红细胞生成素及其受体基因在绵羊各组织中表达量

doi:10.6048/j.issn.1001-4330.2023.02.028

摘要/Abstract

摘要：

【目的】 研究缺氧诱导因子(HIF)及其下游靶基因促红细胞生成素(EPO)及其受体(EPOR)在绵羊正常生理状况下各组织的表达,为靶向调控或缓解绵羊缺氧应激的研究提供组织学依据。【方法】 采用实时荧光定量PCR方法,检测4只新疆细毛羊14种组织中HIF、EPO和EPOR基因的相对表达量。【结果】 HIF-1α和HIF-2α基因均在绵羊肺组织的相对表达量最高,且极显著高于其它组织(P < 0.01);在肺脏HIF-2α基因的表达量约为HIF-1α表达量的5.2倍。EPO基因在绵羊肾脏的相对表达量最高且显著高于垂体、结肠、脾脏、盲肠、肝脏、肾上腺、瘤胃、下丘脑以及心脏(P < 0.05)。EPOR基因在肺脏的表达量最高,肺脏和睾丸的相对表达量显著高于脾脏、下丘脑、肾脏、心脏等组织(P < 0.05)。【结论】 HIF-1α、HIF-2α和EPO及其受体基因在绵羊的肺部或肾脏的高表达,可能是绵羊缺氧应激感受及其调控的重要器官。

关键词: 绵羊; 缺氧诱导因子; 促红细胞生成素; 缺氧应激

Abstract:

【Objective】 To investigate the expression of hypoxia inducible factor (HIF) and its downstream target gene erythropoietin (EPO) and its receptor (EPOR) in ovine tissues under normal physiological conditions.The results have provided a histological basis for the study of targeted regulation or alleviation of ovine hypoxia stress. 【Methods】 The relative expression of HIF, EPO and EPOR genes in 14 tissues of 4 Xinjiang fine wool sheep were detected by real-time fluorescence quantitative PCR. 【Results】 HIF-1α and HIF-2α the relative expression of genes in ovine lung tissues were the highest and significantly higher than those in other tissues (P< 0.01); HIF-2α in lung the gene expression was about HIF-1α 5.2 times.The relative expression of EPO gene in ovine kidney was the highest and significantly higher than that in pituitary, colon, spleen, cecum, liver, adrenal gland, rumen, hypothalamus and heart (P< 0.05).The expression of EPOR gene in lung was the highest, and the relative expression of lung and testis was significantly higher than that in spleen, hypothalamus, kidney and heart (P< 0.05). 【Conclusion】 HIF-1α, HIF-2α and EPO and its receptor genes are highly expressed in the lung or kidney of sheep, suggesting that these two may be important organs for ovine hypoxia stress perception and regulation.

Key words: ovine; hypoxia inducible factor; erythropoietin; hypoxia stress

中图分类号:

S826

雷艳, 兰斌, 余万里, 戴小华, 蔡鹏, 顾伟芳, 阿迪莱·艾力, 赵红琼. 缺氧诱导因子和促红细胞生成素及其受体基因在绵羊各组织中表达量[J]. 新疆农业科学, 2023, 60(2): 485-492.

LEI Yan, LAN Bin, YU Wanli, DAI Xiaohua, CAI Peng, GU Weifang, Adilai Ali, ZHAO Hongqiong. Studies on Expression of Hypoxia Inducible Factor, Erythropoietin Gene and Its Receptor in Ovine Tissues[J]. Xinjiang Agricultural Sciences, 2023, 60(2): 485-492.

图/表 11

表1 采集的绵羊组织样品及位点

Table 1 The samping site of the tissues

组织名称 Sample Name	样品采集位点 Samping site
心脏Heart	右心室心肌
肝脏Liver	右叶边缘
脾脏Spleen	随机
肺脏Lung	右叶边缘
肾脏Kidney	包括皮质和髓质
瘤胃Rumen	后腹盲囊腹侧部
空肠Jejunum	中段
回肠Ileum	距离回盲结口20 cm
结肠Colon	升结肠部位
盲肠Cecum	距离盲端10 cm
十二指肠Duodenum	距幽门口10 cm
睾丸Testis	白膜和睾丸小叶区域
肾上腺Adrenal gland	全部
下丘脑Hypothalamus	全部

表2 HIF-1α、HIF-2α、EPO和GAPDH的PCR引物序列

Table 2 PCR primer sequences of HIF-1α, HIF-2α, EPO and GAPDH

名称 Name	序列 Sequence	长度 Size (bp)
HIF-1a-F	5' ATGAAGTGTACCCTAACTAGCCG 3'	166
HIF-1a-R	5' GTTCACAAATCAGCACCAAGC 3'	166
HIF-2a-F	5' GGACCAGACAGAGTCACCGTT 3'	185
HIF-2a-R	5' TCCCGAAATCCAGAGAAATG 3'	185
EPO-F	5' AGAGGGGACAGGTGACCCAGTGCTA 3'	73
EPO-R	5' ATGGCGTAGGCAGTGATGGTGAGC 3'	73
GAPDH-F	5' GTCGGAGTGAACGGATTTGG 3'	178
GAPDH-R	5' CGTTCTCTGCCTTGACTGTG 3'	178
EPOR-F	5' CAGCCTTGTGGCGATATGGA 3'	139
EPOR-R	5' AGCTGGGGTCAAGAATGGTG 3'	139

图1 绵羊各组织提取的RNA完整性检验

Fig.1 RNA integrity test of ovine tissues

图2 绵羊组织中HIF-1α、HIF-2α、EPO和EPOR基因的扩增

Fig.2 PCR results of HIF-1α, HIF-2α, EPO and EPOR genes from ovine tissues

图3 实时荧光定量PCR熔解曲线

Fig.3 The melting curves of real-time PCR

图4 实时荧光定量PCR扩增曲线

Fig.4 The amplification curves of real-time PCR

图5 HIF-1α基因在绵羊各组织中的相对表达量注:绵羊各组织之间相对表达量具有统计学意义用*表示P < 0.05水平,用**表示P < 0.01水平,下同

Fig.5 Relative expression of HIF-1α gene in ovine tissues Note:The relative expression level among ovine tissues was statistically significant, with * for P < 0.05 and * * for P < 0.01 level,the same as below

图6 HIF-2α基因在绵羊各组织中的相对表达量

Fig.6 Relative expression of HIF-2α gene in ovine tissues

图7 HIF-1α和HIF-2α基因在绵羊肺组织中的相对表达量注:两种基因相对表达量差异具有统计学意义用*表示P < 0.05水平,用**表示P < 0.01水平

Fig.7 Relative expression of HIF-1α and HIF-2α genes in ovine lungs Note:The relative expression level among two genes was statistically significant, with * for P < 0.05 and * * for P < 0.01 level

图8 EPO基因在绵羊各组织中的相对表达量

Fig.8 Relative expression of EPO gene in ovine tissues

图9 EPOR基因在绵羊各组织中的相对表达量

Fig.9 Relative expression of EPOR gene in ovine tissues

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