枸杞多糖对慢性低氧大鼠外周血单个核细胞中HIF-1α、VEGF mRNA和蛋白表达的影响
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摘要
目的探讨慢性低氧对大鼠外周血单个核细胞(PBMCs)中缺氧诱导因子-1α(hypoxia inducible factors-1α,HIF-1α)、血管内皮生长因子(vascular endothelial growth factor,VEGF)表达水平及枸杞多糖(lycium barbarum polysaccharides,LBP)干预的影响。方法 28只Sprague-Dawley(SD)大鼠随机分为常氧组、低氧组、低氧/枸杞多糖组。低氧组设置为常压低氧舱(氧浓度为10%),LBP干预组分别给予LBP[200mg·(kg·d)-1]和[300mg·(kg·d)-1]2个剂量灌胃。各组干预时间均为4周。干预结束后进行血常规检测,然后收集大鼠外周血并分离PBMCs,采用荧光定量PCR和Western blot分别检测大鼠外周血单个核细胞中HIF-1α、VEGF mRNA和蛋白的表达。结果与常氧组相比,低氧组血红蛋白含量和红细胞比容升高(P均<0.01),与低氧组比较,低氧/枸杞多糖(200mg,300mg)组血红蛋白含量和红细胞比容均降低(P<0.05或<0.01);与常氧组相比,低氧组大鼠外周血单个核细胞中HIF-1α、VEGF在mRNA及蛋白表达水平均升高(P均<0.01);与低氧组相比,低氧/枸杞多糖干预的两个剂量组HIF-1α、VEGF mRNA和蛋白表达水平均下降(P<0.05或<0.01);LBP(300mg)组HIF-1α、VEGF mRNA表达低于LBP(200mg)组(P<0.05)。结论慢性低氧引起大鼠PBMCs中HIF-1α和VEGF基因表达水平上调,LBP干预能够在一定程度下调HIF-1α和VEGF基因表达。
Objective To investigate the effects of chronic hypoxia on the expression of hypoxia inducible factors-1α(HIF-1α),vascular endothelial growth factor(VEGF)and Lycium Barum Polysaccharides(LBP)in peripheral blood mononuclear cells(PBMCs)of rats. Methods Total of 28 SD rats were randomly divided into normal control group,hypoxia group,hypoxia/LBP groups(200 and 300 mg·(kg·d)~(-1),respectively). Rats in the hypoxia and hypoxia/LBP groups were subjected to hypoxia exposure in normobaric hypoxia cabin with10% O_2 concentration. At the same time,rats in hypoxia/LBP groups were treated by intragastric administration with LBP[200 mg·(kg·d)~(-1)]and [300 mg·(kg·d)~(-1)],respectively. The intervention was lasted for 4 weeks.Routine blood tests were performed. The peripheral blood was collected and PBMCs was isolated from rats,and the expressions of HIF-1α and VEGF mRNA and protein were detected by fluorescence quantitative PCR and western blotting. Results Compared with the normoxic group,the hemoglobin content and erythrocyte specific volume increased in the hypoxic group(P<0.01),and decreased in the hypoxic/LBP group(200 mg,300 mg)compared with the hypoxic group(P<0.05 or <0.01). Compared with the normoxia group,the mRNA and protein expression levels of HIF-1α and VEGF in peripheral blood mononuclear cells of hypoxic rats were increased(P<0.01). Compared with the hypoxic group,mRNA and protein expression levels of HIF-1α and VEGF in both dose groups(200 mg group,300 mg group)of hypoxia/LBP decreased(P<0.05 or <0.01). The mRNA expression of HIF-1α and VEGF in 300 mg group of LBP concentration was lower than that in 200 mg group of LBP concentration(P<0.05). Conclusion Chronic hypoxia induced the up-regulation of HIF-1 αand VEGF gene expression in PBMCs of rats. LBP intervention could down-regulate the expression of HIF-1αand VEGF gene to a certain extent.
Objective To investigate the effects of chronic hypoxia on the expression of hypoxia inducible factors-1α(HIF-1α),vascular endothelial growth factor(VEGF)and Lycium Barum Polysaccharides(LBP)in peripheral blood mononuclear cells(PBMCs)of rats. Methods Total of 28 SD rats were randomly divided into normal control group,hypoxia group,hypoxia/LBP groups(200 and 300 mg·(kg·d)~(-1),respectively). Rats in the hypoxia and hypoxia/LBP groups were subjected to hypoxia exposure in normobaric hypoxia cabin with10% O_2 concentration. At the same time,rats in hypoxia/LBP groups were treated by intragastric administration with LBP[200 mg·(kg·d)~(-1)]and [300 mg·(kg·d)~(-1)],respectively. The intervention was lasted for 4 weeks.Routine blood tests were performed. The peripheral blood was collected and PBMCs was isolated from rats,and the expressions of HIF-1α and VEGF mRNA and protein were detected by fluorescence quantitative PCR and western blotting. Results Compared with the normoxic group,the hemoglobin content and erythrocyte specific volume increased in the hypoxic group(P<0.01),and decreased in the hypoxic/LBP group(200 mg,300 mg)compared with the hypoxic group(P<0.05 or <0.01). Compared with the normoxia group,the mRNA and protein expression levels of HIF-1α and VEGF in peripheral blood mononuclear cells of hypoxic rats were increased(P<0.01). Compared with the hypoxic group,mRNA and protein expression levels of HIF-1α and VEGF in both dose groups(200 mg group,300 mg group)of hypoxia/LBP decreased(P<0.05 or <0.01). The mRNA expression of HIF-1α and VEGF in 300 mg group of LBP concentration was lower than that in 200 mg group of LBP concentration(P<0.05). Conclusion Chronic hypoxia induced the up-regulation of HIF-1 αand VEGF gene expression in PBMCs of rats. LBP intervention could down-regulate the expression of HIF-1αand VEGF gene to a certain extent.
引文
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[13]Ivan M,Kondo K,Yang H,et al.HIFαtargeted for VHL-mediated destruction by proline hydroxylation:implications for O2sensing[J].Science,2001,292:464-468.
[14]Kaelin WG,Ratcliffe PJ.Oxygen sensing by metazoans:The central role of the HIF hydroxylase pathway[J].Mol Cell,2008,30:393-402.
[15]Haase VH.Hypoxic regulation of erythropoiesis and iron metabolism[J].Am J Physiol Renal Physiol,2010,299(1):F1.
[16]Suzuki N,Obara N,Pan X,et al.Specific contribution of the erythropoietin gene 3'enhancer to hepatic erythropoiesis after late embryonic stages[J].Molecular&Cellular Biology,2011,31(18):3896.
[17]李芳,关文霞,任飞,等.Sirt1和HIF-1α在慢性阻塞性肺疾病患者外周血单个核细胞中的表达及其意义[J].吉林大学学报,2015,41(2):356-361.
[18]王宗强,于荣秀,吴健红,等.缺氧诱导因子-1α在COPD患者外周血单个核细胞中的表达[J].广东医学,2011,32(1):64-66.
[19]Bates DO.Vascular endothelial growth factors and vascular permeability[J].Cardiovasc Res,2010,87(2):262-271.
[20]Yamazaki Y,Egawa K,Nose K,et al.HIF-1-dependent VEGF reporter gene assay by a stable transformant of CHO cells[J].Biological&Pharmaceutical Bulletin,2003,26(4):417-420.
[2]Fan X,Heijnen CJ,Ma VDK,et al.The role and regulation of hypoxia-inducible factor-1alpha expression in brain development and neonatal hypoxic-ischemic brain injury[J].Brain Research Reviews,2009,62(1):99-108.
[3]冯彦.枸杞多糖药理作用与临床应用[J].中国现代药物应用,2016,10(6):278-279.
[4]齐春会,张永祥,赵修南,等.枸杞粗多糖的免疫活性[J].中国药理学与毒理学杂志,2001,(3):180-184.
[5]Luo Q,Cai Y,Yan J,et al.Hypoglycemic and hypolipidemic effects and antioxidant activity of fruit extracts from Lycium barbarum[J].Life Sci,2004,76(2):137-149.
[6]王琦,张晓丹,张静蕾.枸杞多糖研究进展[J].食品研究与开发,2009,30(10):150-152.
[7]冉丕鑫,段生福.常压缺氧性大鼠肺动脉高压模型的建立[J].衡阳医学院学报,1994,22(3):229-231.
[8]Semenza GL,Agani F,Feldser D,et al.Hypoxia,HIF-1,and the pathophysiology of common human diseases[J].Oxygen Transport to Tissue XXXIII,2000,475:123-130.
[9]Gassmann M.The impact of hypoxia on our body:from integrative physiology to human disease[J].Cell Mol Life Sci,2009,66(22):3537-3538.
[10]Ratcliffe PJ.Oxygen sensing and hypoxia signalling pathways in animals:The implications of physiology for cancer[J].Physiol,2013,591:2027-2042.
[11]Semenza GL.Hydroxylation of HIF-1:Oxygen sensing at the molecular level[J].Physiology(Bethesda),2004,19:176-182.
[12]Yamaguchi J,Tanaka T,Eto N,et al.Inflammation and hypoxia linked to renal injury by CCAAT/enhancer-binding protein[J].Kidney International,2015,88(2):262-275.
[13]Ivan M,Kondo K,Yang H,et al.HIFαtargeted for VHL-mediated destruction by proline hydroxylation:implications for O2sensing[J].Science,2001,292:464-468.
[14]Kaelin WG,Ratcliffe PJ.Oxygen sensing by metazoans:The central role of the HIF hydroxylase pathway[J].Mol Cell,2008,30:393-402.
[15]Haase VH.Hypoxic regulation of erythropoiesis and iron metabolism[J].Am J Physiol Renal Physiol,2010,299(1):F1.
[16]Suzuki N,Obara N,Pan X,et al.Specific contribution of the erythropoietin gene 3'enhancer to hepatic erythropoiesis after late embryonic stages[J].Molecular&Cellular Biology,2011,31(18):3896.
[17]李芳,关文霞,任飞,等.Sirt1和HIF-1α在慢性阻塞性肺疾病患者外周血单个核细胞中的表达及其意义[J].吉林大学学报,2015,41(2):356-361.
[18]王宗强,于荣秀,吴健红,等.缺氧诱导因子-1α在COPD患者外周血单个核细胞中的表达[J].广东医学,2011,32(1):64-66.
[19]Bates DO.Vascular endothelial growth factors and vascular permeability[J].Cardiovasc Res,2010,87(2):262-271.
[20]Yamazaki Y,Egawa K,Nose K,et al.HIF-1-dependent VEGF reporter gene assay by a stable transformant of CHO cells[J].Biological&Pharmaceutical Bulletin,2003,26(4):417-420.