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码上扫一扫！ |
| 藏猪的低氧适应性机制研究 |
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王语诗1,2, 吴海彬3, 泽里磋1,2, 张非凡1,2, 吴茜1,2, 韦明邦1,2, 张健1,2, 强巴央宗1,2, 商鹏1,2
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1.西藏农牧学院 动物科学学院，西藏 林芝860000;2.藏猪遗传改良与繁殖工程重点实验室,西藏 林芝860000;3.西藏自治区兽医生物药品制造厂,西藏 拉萨 850000
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| 摘要: |
| 【目的】探究Ⅲ型纤连蛋白域包含蛋白5（type Ⅲ fibronectin domain-containing protein 5，FNDC5）基因对藏猪低氧适应性的调控作用。【方法】以藏猪和大约克猪为实验动物，提取其耳组织DNA，分段扩增FNDC5基因的5′侧翼区、3′侧翼区和CDS区，测序后筛选单核苷酸多态性（SNPs）位点。采集实验猪心脏、肺脏组织，提取其RNA及总蛋白，采用实时荧光定量PCR（RT-qPCR）法检测FNDC5基因mRNA的表达量，采用Western blot法检测FNDC5蛋白的表达量。采集藏猪和大约克猪颈动脉血样各30头份，测定血液生理指标。将猪骨骼肌卫星细胞（PSMSC）分为常氧组（氧气体积分数21%）及体积分数5%和1%氧气组，常氧组又分为PGC 1α激活剂ZLN005添加组、PGC-1α抑制剂SR-18292添加组和空白对照组，培养后分别采用RT-qPCR和Western blot法检测FNDC5基因mRNA和蛋白的表达量。【结果】在FNDC5基因CDS区,2种猪未检测到SNPs位点；在5′侧翼区，2种猪有3个SNPs位点，分别为g.-118A>G、g.-790T>C和g.-931A>G；在3′侧翼区，大约克猪无SNP位点，藏猪有4个多态性位点（分别为g.46G>T、g.56G>C、g.517T>C和g.1028G>A）。这些SNPs可能是藏猪低氧适应性的重要调控位点。藏猪的白细胞数、红细胞数、血红蛋白质量浓度、红细胞压积、血小板计数、血小板压积指标极显著高于大约克猪（P＜0.01），红细胞分布宽度变异系数极显著低于大约克猪（P＜0.01），其他指标与大约克猪无显著差异，推测藏猪具有更强的低氧适应能力。藏猪心脏FNDC5基因和蛋白表达量极显著高于大约克猪，肺脏FNDC5基因表达量极显著高于大约克猪。在PSMSC中，FNDC5基因和蛋白的表达量均随着氧体积分数的降低而显著或极显著降低；PGC-1α激活剂ZLN005可极显著促进FNDC5基因的表达，抑制剂SR-18292可极显著抑制FNDC5基因的表达。【结论】FNDC5基因和部分血液生理指标可能与藏猪的低氧适应性相关；PGC-1α可正向调控FNDC5基因的表达。 |
| 关键词: 藏猪 FNDC5基因 低氧适应性 单核苷酸多态性 |
| DOI：DOI:10.13207/j.cnki.jnwafu.2025.06.001 |
| 分类号: |
| 基金项目:西藏自治区科技计划项目（XZ202202JD0002N）;国家重点研发计划项目（2022YFD1600903）;西藏自治区重大科技专项项目（XZ202101ZD0005N）；西藏农牧学院研究生教育创新计划项目（YJS2023-14） |
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| Studies on the mechanism of hypoxia adaptation in Tibetan pigs |
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WANG Yushi1,2, WU Haibin3, ZE Licuo1,2, ZHANG Feifan1,2, WU Xi1,2, WEI Mingbang1,2, ZHANG Jian1,2, CHAMBA Yangzom1,2, ZHANG Jian1,2
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1.College of Animal Science，Tibet Agricultural and Animal Husbandry University,Linzhi,Xizang 860000,China;2.Tibetan Pig Genetic Improvement and Reproduction Engineering Key Laboratory,Linzhi,Xizang 860000,China;3.Tibet Autonomous Region Veterinary Biological Drug Manufacturing Factory,Lhasa,Xizang 850000,China
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| Abstract: |
| 【Objective】This research aimed to investigate the regulation effect of type Ⅲ fibronectin domain-containing protein 5 (FNDC5) gene on hypoxic acclimatization of Tibetan pigs.【Method】Tibetan pigs and Yorkshire pig were used as experimental animals,and their ear tissue DNA was extracted.The 5′ flanking region,3′ flanking region and CDS region of the FNDC5 gene were amplified in segments,and their sequences were determined to screen single nucleotide polymorphism sites (SNPs).Heart and lung tissues of experimental pigs were collected,and RNA and total protein of heart and lung tissues were extracted.The expression of mRNA of FNDC5 gene was detected by real time fluorescence quantitative PCR (RT-qPCR) method,and the expression of FNDC5 protein was detected by Western blot.In addition,carotid artery blood samples of 5 mL were collected from each of Tibetan pigs(30) and Yorkshire pigs(30) for the determination of blood physiological indexes.Porcine skeletal muscle satellite cells (PSMSC) were divided into normoxia concentration group (21%),5% oxygen concentration group and 1% oxygen concentration group,and the normoxia concentration group was further divided into the group with the addition of the PGC-1α activator ZLN005,the group with the addition of the PGC 1α inhibitor SR-18292,and the control group.They were cultured and then detected by RT-qPCR and Western blot to determine their FNDC5 gene mRNA and protein expression level,respectively.【Result】No SNP sites were detected in the CDS region of the FNDC5 gene of the two pig species,and there were three SNP sites in the 5′ flanking region of the two pig species,namely g.－118A>G,g.－790T>C,g.－931A>G.There were no SNP sites in the 3′ flanking region of the FNDC5 gene of the Yorkshire pig,and there were four polymorphic sites in the Tibetan pig,namely g.46G>T,g.56G>C,g.517T>C,and g.1028G>A.These SNPs may be important regulatory loci for hypoxia adaptation in Tibetan pigs.The leukocyte count,erythrocyte count,hemoglobin content, erythrocyte specific area,platelet count,platelet specific area indexes of Tibetan pigs were highly significantly higher than those of Yorkshire pig (P＜0.01),and the coefficient of variation for the width of erythrocyte distribution was highly significantly lower than that of Yorkshire pig (P＜0.01).No significant difference was observed between other indexes of Yorkshire pig and Tibetan pigs.Therefore,it was hypothesized that the Tibetan pigs had a better hypoxic adaptive ability.The expression level of FNDC5 gene and protein in the heart of Tibetan pigs was highly significantly higher than those of Yorkshire pig,and the expression level of FNDC5 gene in the lungs was highly significantly higher than that of Yorkshire pig.In PSMSC,the expression level of FNDC5 gene and protein decreased significantly or highly significantly with the decrease of oxygen concentration.The PGC-1α activator ZLN005 promoted the expression of FNDC5 gene highly significantly,and the inhibitor SR-18292 inhibited the expression of FNDC5 gene highly significantly.【Conclusion】The FNDC5 gene and some blood physiological indices may be associated with the hypoxic acclimatization in Tibetan pigs,and PGC-1α positively regulates the expression of the FNDC5 gene. |
| Key words: Tibetan pig FNDC5 gene hypoxia acclimatization single nucleotide poly morphism |
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