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

Effects of Vitamin C on the Transcription of Hypoxia Inducible Factor-1α of Broiler Pulmonary Artery Smooth Muscle Cells under Hypoxia

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21d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 500μmol·L-1),5个培养时间(6、12、24、48和72h)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,250μmol·L-1 CoCl2处理48h可成功建立肉鸡PASMCs缺氧模型;500或1 000μmol·L-1VC处理48或72h可显著降低缺氧PASMCs中HIF-1α、VEGFR2mRNA的转录(P<0.05),而1 500μmol·L-1VC则显著增加缺氧PASMCs中HIF-1α、VEGF、VEGFR2mRNA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。

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 7different concentrations of VC（0,50,100,200,500,1 000 and 1 500μmol·L^-1）and 5processing time（6,12,24,48 and 72h）,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 htreatment byCoCl2with 250μmol·L^-1（P〈0.05）.The relative transcription of HIF-1α,VEGFR2 in hypoxia PASMCs were significantly decreased by VC with 500 or 1 000μmol·L^-1 after 48 or 72h（P〈0.05）.Whereas 1 500μmol·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 upregulate 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.