摘要
目的:探讨葛根素(puerarin,PUE)预处理对缺氧/复氧(hypoxia reoxygenation,H/R)损伤的人脐静脉内皮细胞(HUVECs)的保护作用。方法:HUVECs随机分为正常对照(control)组、H/R组、单纯PUE组和PUE+H/R组(1.0×10^(-3)mol/L PUE预处理24 h后进行H/R)。采用Western blot方法检测内皮型一氧化氮合酶(e NOS)蛋白的表达,化学比色法检测组成型一氧化氮合酶(c NOS)的活性,TUNEL法检测细胞凋亡。此外,在PUE预处理前使用ERK蛋白激酶抑制剂U0126(1.0×10^(-5)mol/L)或PI3K/Akt蛋白激酶抑制剂LY294002(5.0×10^(-5)mol/L)处理细胞1 h,再进行H/R。结果:与control组相比,H/R组的e NOS蛋白表达水平降低(P<0.05),PUE预处理上调e NOS蛋白的表达水平(P<0.05),该上调作用均可被U0126及LY294002抑制(P<0.05);与control组相比,H/R组的c NOS活力下降(P<0.05),PUE预处理组的c NOS活力增加(P<0.05);与control组相比,H/R组细胞凋亡指数显著增大(P<0.01),PUE预处理组的细胞凋亡指数减小(P<0.01)。结论:H/R可使HUVECs受损伤,e NOS蛋白表达及活性降低,细胞凋亡增加。PUE预处理可通过ERK1/2信号通路和PI3K/Akt信号通路上调HUVECs的e NOS蛋白表达,增强e NOS活性,减少细胞凋亡,发挥内皮细胞保护作用。
AIM： To investigate the protective effects of puerarin（ PUE） pretreatment on hypoxia/reoxygenation（ H/R） injury in human umbilical vein endothelial cells（ HUVECs）,as well as its possible mechanism and the signal transduction pathways involved. METHODS： HUVECs were randomly divided into normal control group,H / R group,PUE pretreatment group and PUE ＋ H / R group（ 1. 0 × 10^（-3）mol / L,PUE pretreated the cells for 24 h before H / R）. The protein expression of endothelial nitric oxide synthase（ e NOS） was measured by Western blot. The activity of constitutive NOS（ c NOS） was determined via chemical colorimetric methods. Apoptosis of HUVECs was detected by TUNEL assay. In addition,the cells were treated with ERK inhibitor U0126（ 1. 0 × 10^（-5）mol / L） or PKB / Akt inhibitor LY294002（ 5. 0 × 10^（-5）mol / L） for 1 h before PUE pretreatment,and then H / R was performed. RESULTS： Compared with control group,H / R decreased the protein expression of e NOS（ P〈0. 05）,and PUE pretreatment up-regulated it（ P〈0. 05）. This effect of PUE was inhibited by U0126 or LY294002（ P〈0. 05）. Compared with control group,the activity of c NOS decreased in H / R group（ P〈0. 05）,while it increased after PUE pretreatment（ P〈0. 05）. Compared with control group,the apoptotic index significantly increased in H / R group（ P〈0. 01）. PUE pretreatment reduced the apoptotic index（ P〈0. 01）. CONCLUSION： H / R decreases the protein expression and enzyme activity of e NOS in HUVECs,and induces apoptosis of HUVECs.PUE pretreatment up-regulates the protein expression and enzyme activity of e NOS,and reduces the apoptosis of HUVECs with H / R injury. The protective effect of PUE might be through increasing e NOS protein expression via ERK1 /2 and PKB /Akt signaling pathways.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2016年第5期857-862,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.31460267)
贵州省社会发展攻关项目[黔科合SY字(2013)3019号]
关键词
葛根素
人脐静脉内皮细胞
缺氧/复氧
内皮型一氧化氮合酶
Puerarin
Human umbilical vein endothelial cells
Hypoxia/reoxygenation
Endothelial nitric oxide synthase
