摘要
目的观察血管紧张素Ⅱ(AngⅡ)对人脐动脉平滑肌细胞(HUASMC)缺氧诱导因子1α(HIF-1α)、血管内皮生长因子(VEGF)以及脯氨酸羟化酶2(PHD2)、缺氧诱导因子1抑制因子(FIH-1)、p-ERK表达的影响,探讨PHD2、FIH-1、p-ERK在AngⅡ作用下对HIF-1α表达的影响机制。方法 HUASMC分为三组:(1)对照组:正常培养基培养细胞6 h;(2)AngⅡ组:含AngⅡ的培养基(终浓度为10-6mol/L)培养细胞6 h;(3)AngⅡ+PD98059组:含PD98059的培养基(终浓度为10-5mol/L)预处理细胞1 h后,加入含AngⅡ的培养基(终浓度为10-6mol/L)培养细胞6 h。RT-PCR检测细胞HIF-1α、VEGF、PHD2、FIH-1的基因表达;Western blot检测上述目的蛋白表达及p-ERK表达。结果 (1)与对照组比较,AngⅡ增加HUASMC中HIF-1α、VEGF基因和蛋白表达(P<0.05),增加pERK蛋白表达(P<0.05),降低FIH-1基因和蛋白表达(P<0.05),不影响PHD2基因和蛋白表达;(2)与AngⅡ组比较,ERK抑制剂PD98059降低HUASMC中HIF-1α、VEGF基因和蛋白的表达(P<0.05)以及p-ERK蛋白的表达(P<0.05),增加FIH-1的基因和蛋白表达(P<0.05)。结论 (1)AngⅡ增加HUASMC中HIF-1α、VEGF的表达,该作用通过激活ERK通路、抑制FIH-1表达,在翻译后水平减少HIF-1α的降解所致;(2)ERK抑制剂可削弱AngⅡ的促HIF-1α、VEGF表达作用。
Aim To observe the affection of angiotensin Ⅱ（ AngⅡ） on hypoxia-inducible factor-1（ HIF-1α）,vascular endothelial growth factor（ VEGF）,prolyl hydroxylases-2（ PHD2）,factor inhibiting hypoxia-inducible factor-1（ FIH-1）,and p-ERK expression in human umbilical artery smooth muscle cells（ HUASMC）,and clarify the mechanism of PHD2,FIH-1,and p-ERK on HIF-1α expression on the condition of AngⅡ. Method HUASMC were divided into：（ 1） Control group： normal culture medium for 6 hours;（ 2） AngⅡ group： 10- 6mol /L AngⅡ culture medium for 6hours;（ 3） AngⅡ ＋ PD98059 group： 10- 5mol /L PD98059 added 1 hour before 10- 6mol /L AngⅡ,and then for 6 hours.Gene expression of HIF-1α,VEGF,PHD2 and FIH-1 were checked by real-time PCR,and the corresponding proteins of above factors,and the p-ERK activation were checked with Western blot. Results（ 1） Compared with control group,AngⅡ promoted both gene and protein expression of HIF-1α and VEGF（ P〈0. 05）,and activated p-ERK（ P〈0. 05）,with the decreased FIH-1 gene and protein expression（ P〈0. 05）,but has no effect on PHD2 gene and protein expression in HUASMC.（ 2） Compared with AngⅡ group,both gene and protein expression of HIF-1α,VEGF and the p-ERK activation were significantly reduced（ P〈0. 05）,with the increased gene and protein expression of FIH-1（ P〈0. 05） in HUASMC. Conclusions（ 1） AngⅡ promoted both gene and protein expression of HIF-1α and VEGF in HUASMC.Its mechanism is that activated ERK pathway inhibited FIH-1,leading post-translational reduction of degradation of HIF-1α.（ 2） ERK inhibitor weakened the promoted affection of AngⅡ on HIF-1α and VEGF.
出处
《中国动脉硬化杂志》
CAS
CSCD
北大核心
2014年第8期799-802,共4页
Chinese Journal of Arteriosclerosis
