牦牛Nramp1基因mRNA组织表达谱及其相关miRNAs初步研究

doi:10.11843/j.issn.0366-6964.2019.08.006

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (8): 1576-1586.doi: 10.11843/j.issn.0366-6964.2019.08.006

牦牛Nramp1基因mRNA组织表达谱及其相关miRNAs初步研究

温东旭^1,2, 张雷¹, 索朗斯珠^1,2, 牛家强^1,2, 王玉恒³, 贡嘎^1,2, 旦巴次仁¹, 席广银⁴, 郭敏⁴, 徐业芬^1,2*

1. 西藏农牧学院动物科学学院, 林芝 860000;
2. 西藏农牧学院西藏高原动物疫病研究自治区高校重点实验室, 林芝 860000;
3. 西藏自治区兽医生物药品制造厂, 拉萨 850000;
4. 中国农业大学动物科技学院, 北京 100193

收稿日期:2019-01-27 出版日期:2019-08-23 发布日期:2019-08-19
通讯作者: 徐业芬,主要从事高原动物免疫遗传和生理学研究,E-mail:xzlzxyf@163.com
作者简介:温东旭(1993-),男,内蒙古太仆寺旗人,硕士生,主要从事高原动物免疫遗传研究,E-mail:804223267@qq.com;张雷(1995-),男,甘肃定西人,本科生,E-mail:2191949433@qq.com。温东旭和张雷为同等贡献作者
基金资助:
西藏农牧学院雪域英才青年拔尖人才项目（2019-6）；国家肉牛牦牛产业体系项目（CARS-37）；西藏自治区大学生创新性实验项目（2019-14）

Preliminary Study on Tissue Expression Profile and Related miRNAs of Yak Nramp1 Gene

WEN Dongxu^1,2, ZHANG Lei¹, SUOLANG Sizhu^1,2, NIU Jiaqiang^1,2, WANG Yuheng³, GONG Ga^1,2, DANBA Ciren¹, XI Guangyin⁴, GUO Min⁴, XU Yefen^1,2*

1. Department of Animal Science, Tibet Agriculture & Animal Husbandry University, Linzhi 860000, China;
2. Provincial Key Laboratory of Tibet Plateau Animal Epidemic Disease Research, Tibet Agriculture & Animal Husbandry University, Linzhi 860000, China;
3. Tibet Veterinary Biological Pharmaceutical Factory, Lhasa 850000, China;
4. College of Animal Science and Technology, China Agricultural University, Beijing 100193, China

Received:2019-01-27 Online:2019-08-23 Published:2019-08-19

摘要/Abstract

摘要： 旨在探究牦牛Nramp1基因mRNA及可能靶定Nramp1的miRNAs组织表达谱。本研究利用RT-PCR技术对牦牛Nramp1基因的mRNA组织表达谱进行分析，同时运用TargentScan和miRBase软件预测牦牛可能靶定Nramp1基因的miRNAs，并利用加PloyA尾法RT-PCR技术分析miRNAs在肝、脾、肺、肾、后腿骨骼肌、卵巢、小肠、颌下淋巴结、大肠、肠系淋巴结10种组织中的相对表达量。试验结果显示，Nramp1基因mRNA在所检测的10种组织中均有表达，其中在脾、颌下淋巴结及肺组织中的表达量极显著高于肝、肾、大肠、肠系淋巴结组织（P<0.01），显著高于小肠组织表达量（P<0.05）。预测到可能靶定牦牛Nramp1基因的miRNAs共有201个，从中选择6个进行表达谱分析，发现bta-miR-106a、bta-miR-20b、bta-miR-17-5p在牦牛免疫组织脾、颌下淋巴结、肠系淋巴结中，bta-miR-93、bta-miR-106b、bta-miR-20a在牦牛免疫组织颌下淋巴结、肠系淋巴结中均与Nramp1基因mRNA共表达，但是bta-miR-93、bta-miR-20a、bta-miR-106a、bta-miR-17-5p和bta-miR-20b在颌下淋巴结和肠系淋巴结间的表达量无显著差异（P>0.05），而bta-miR-106a、bta-miR-20b在颌下淋巴结、肠系淋巴结组织中的表达量极显著高于脾组织表达量（P<0.01），bta-miR-106b在颌下淋巴结组织中的表达量极显著高于肠系淋巴结组织表达量（P<0.01）。Nramp1基因mRNA在牦牛体内广泛表达说明其可能具有广泛免疫调节作用；bta-miR-93、bta-miR-20b、bta-miR-106a、bta-miR-106b、bta-miR-20a和bta-miR-17-5p与Nramp1基因mRNA在免疫组织中的共表达表明二者可能具有靶向调控关系参与免疫调控作用，但二者是否存在真实的靶向调控以及其调控机制有待于深入研究。

关键词: 牦牛, Nramp1基因, miRNA, 组织表达谱

Abstract: The aim of this study was to investigate the tissue expression profiles of yak Nramp1 gene mRNA and the possible miRNAs targeting to Nramp1. The mRNA expression profile of yak Nramp1 gene was analyzed by RT-PCR, and the possible miRNAs targeting to Nramp1 were predicted by TargentScan and miRBase softwares, and the relative expression levels for these miRNAs were analyzed in the 10 yak tissues including liver, spleen, lung, kidney, hind leg skeletal muscle, ovary, small intestine, submandibular lymph node, large intestine and intestinal lymph nodes by adding PloyA tail RT-PCR method. The results showed that the Nramp1 gene mRNA was expressed in the 10 examined tissues, and the expression levels in the yak spleen, submandibular lymph node and lung were extremely significantly higher than that in the liver, kidney, large intestine and intestinal lymph node(P<0.01), and significantly higher than that in small intestine tissue (P<0.05). A total of 201 miRNAs that might target to the yak Nramp1 gene were predicted, and 6 of them were selected for expression profiling analysis. It was found that Nramp1 mRNA and bta-miR-106a, bta-miR-20b, bta-miR-17-5p were co-expressed in the yak immune tissues such as the spleen, submandibular lymph node and intestinal lymph node, and were co-expressed with bta-miR-93, bta-miR-106b and bta-miR-20a in yak submandibular lymph nodes and intestinal lymph nodes. There were no significant differences in the expression of bta-miR-93,bta-miR-20a,bta-miR-106a,bta-miR-17-5p and bta-miR-20b between submandibular lymph nodes and intestinal lymph nodes(P>0.05), but the expression of bta-miR-106a and bta-miR-20b in submandibular lymph nodes and intestinal lymph nodes were extremely significantly higher than those in spleen tissues(P<0.01),and the expression of bta-miR-106b in submandibular lymph nodes was extremely significantly higher than that in intestinal lymph nodes(P<0.01). The extensive expression of Nramp1 mRNA in various tissues of yak suggested that it might have a wide range of immunomodulatory effects, and the co-expression of bta-miR-93, bta-miR-20b, bta-miR-106a, bta-miR-106b, bta-miR-20a, bta-miR-17-5p and Nramp1 mRNA in immune tissues indicated that they might have targeted regulatory relationship in immune regulation, but whether there is targeting regulation and their regulation mechanism remains to be further studied.

Key words: yak, Nramp1 gene, miRNA, tissue expression profile

中图分类号:

S823.2

温东旭, 张雷, 索朗斯珠, 牛家强, 王玉恒, 贡嘎, 旦巴次仁, 席广银, 郭敏, 徐业芬. 牦牛Nramp1基因mRNA组织表达谱及其相关miRNAs初步研究[J]. 畜牧兽医学报, 2019, 50(8): 1576-1586.

WEN Dongxu, ZHANG Lei, SUOLANG Sizhu, NIU Jiaqiang, WANG Yuheng, GONG Ga, DANBA Ciren, XI Guangyin, GUO Min, XU Yefen. Preliminary Study on Tissue Expression Profile and Related miRNAs of Yak Nramp1 Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(8): 1576-1586.

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牦牛Nramp1基因mRNA组织表达谱及其相关miRNAs初步研究

Preliminary Study on Tissue Expression Profile and Related miRNAs of Yak Nramp1 Gene

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