维生素C对肉鸡缺氧肺动脉平滑肌细胞缺氧诱导因子-1α转录水平的影响

doi:10.11843/j.issn.0366-6964.2016.02.024

摘要/Abstract

摘要：

旨在体外培养肉鸡肺动脉平滑肌细胞(PASMCs)的基础上利用氯化钴（CoCl2）建立细胞缺氧模型，探讨维生素C（VC）对缺氧肉鸡PASMCs中缺氧诱导因子-1α（HIF-1α）及其下游靶基因血管内皮生长因子（VEGF）及血管内皮生长因子受体2（VEGFR2）mRNA转录的影响。选用21 d健康AA肉公鸡，分离、培养并鉴定PASMCs，利用CoCl₂建立细胞缺氧模型，再用7个浓度的VC（0、50、100、200、500、1 000和1 500 μmol•L^-1），5个培养时间（6、12、24、48和72 h）处理，试验共设35个处理，每个处理6个重复。结果表明，利用组织块法可成功培养出肉鸡PASMCs；与对照组相比，不同浓度CoCl₂均可使细胞缺氧基因（HIF-1α、VEGF、VEGFR2）转录量显著增加（P<0.05），促进细胞增殖，250 μmol•L^-1 CoCl₂处理48 h可成功建立肉鸡PASMCs缺氧模型；500或1 000 μmol•L^-1 VC处理48或72 h可显著降低缺氧PASMCs中HIF-1α、VEGFR2 mRNA的转录（P<0.05），而1 500 μmol•L^-1 VC则显著增加缺氧PASMCs中HIF-1α、VEGF、VEGFR2 mRNA的转录（P<0.05）。结果提示，缺氧会导致肉鸡肺动脉平滑肌细胞缺氧基因表达量与细胞增殖增加，而VC能降低细胞对缺氧信号的敏感性，使缺氧基因的相对表达量降低。

Abstract:

This study was conducted to investigate the effects of Vitamin C (VC) on hypoxia inducible factor-1α (HIF-1α)，vascular endothelial growth factor (VEGF) and its receptor 2 (VEGFR2) mRNA transcription of broiler pulmonary artery smooth muscle cells（PASMCs）under hypoxia induced by CoCl₂.PASMCs from healthy AA broilers at 21 days of age were isolated，cultured and identified.A cell model of hypoxia was established by using CoCl₂.On this basis，PASMCs were treated by 7 different concentrations of VC (0，50，100，200，500，1 000 and 1 500 μmol•L^-1) and 5 processing time (6，12，24，48 and 72 h)，total 35 treatments with six replicates.The results indicated that the PASMCs of broilers were successfully separated and cultured in vitro.As for hypoxia model of PASMCs，the relative transcription of HIF-1α，VEGF，VEGFR2 and cell proliferation capacity significantly increased when PASMCs were after 48 h treatment by CoCl₂with 250 μmol•L^-1 (P<0.05).The relative transcription of HIF-1α，VEGFR2 in hypoxia PASMCs were significantly decreased by VC with 500 or 1 000 μmol•L^-1 after 48 or 72 h (P<0.05).Whereas 1 500 μmol•L^-1 VC significantly increased the relative transcription of HIF-1α，VEGF，VEGFR2 in hypoxia PASMCs (P<0.05).These results suggested that anoxia could up-regulate the relative expression of hypoxia related genes and induce abnormally proliferation of PASMCs.While VC could reduce the sensitivity to oxygen signals of hypoxia PASMCs，and further down-regulate the relative expression of hypoxia related genes.

中图分类号:

S852.2

曹林，曾秋凤，张克英，丁雪梅，白世平，罗玉衡，王建萍，玄玥，宿卓薇. 维生素C对肉鸡缺氧肺动脉平滑肌细胞缺氧诱导因子-1α转录水平的影响[J]. 畜牧兽医学报, 2016, 47(2): 388-396.

CAO Lin，ZENG Qiu-feng，ZHANG Ke-ying，DING Xue-mei，BAI Shi-ping，LUO Yu-heng，WANG Jian-ping，XUAN Yue，SU Zhuo-wei. Effects of Vitamin C on the Transcription of Hypoxia Inducible Factor-1α of Broiler Pulmonary Artery Smooth Muscle Cells under Hypoxia[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(2): 388-396.

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