腺苷酸环化酶及蛋白激酶A在β受体激动增加eNOS活性信号通路中的作用  

Essential Role of Adenylyl Cyclase and Protein Kinase-A for Endothelial Nitric Oxide Synthase Activation by β-adrenoceptors Stimulation (β-AR)

作　　者：蒲娟娟[1] 李涛 

机构地区：[1]郑州大学第一附属医院老年病科,450052 [2]河南省高等学校临床医学重点学科开放实验室

出　　处：《中国实用医药》2008年第18期15-16,共2页China Practical Medicine

摘　　要：目的阐明β受体激动增加内皮型一氧化氮合酶(eNOS)活性信号通路中腺苷酸环化酶(AC)及蛋白激酶A(PKA)的作用。方法体外培养人脐静脉内皮细胞;运用同位素两步色谱法([3H]-L-精氨酸转化法)检测eNOS活性。本研究设置空白对照组、AC组和PKA组,每组因素下再分ISO及BSS两个干预因素组,观察AC特异性阻断剂SQ-2256(5×10-5mol/L)及PKA特异性阻断剂H-89(1×10-7mol/L)分别与人脐静脉内皮细胞孵育10min,再与异丙肾上腺素(ISO)孵育30min后eNOS活性变化。结果ISO明显增加eNOS活性(30.7±3.9)%,P<0.01;分别阻断AC、PKA,ISO诱导的eNOS活性增加均受到抑制(12.1±3.53)%、(4.73±2.19)%,P<0.01。结论ISO激活eNOS活性的信号通路上,AC和PKA均有参与。Objective To elucidate the role of adenylyl cyclase （AC）and cyclic AMP-dependent protein kinase-A （PKA）in the signal transduction of the endothelial nitric oxide synthase（eNOS） activation by βAR stimulation in human umbilical vein endothelial cell（HUVEC）. Methods HUVEC was isolated and cultured; HUVEC was added SQ-22536（5 ×10^-5 mol/L,AC inhibitor）and H-89（ 1×10^-7 mol/L,PKA inhibitor）separately for 10 min,then agonist isoprenaline（1×10^-6 mol/L）or vehicle was added and the incubation continued for another 30 min; Evaluation of NOS activity was determined by the conversion of L-[ 3H] arginine to L-[ 3H ] citrulline. Results Isoprenaline increased [3H] L-citrulline formation by（30.7 ±3.9）% （P 〈0.001 vs control）. This increase was significantly inhibited by SQ-22536 and H89. Percentage increases above control were （ 12. 1± 3.53 ） %, （4.73 ± 2. 19） % （ P 〈 0. 001 vs ISO ）. Conclusion AC and PKA are involved in the signal transduction of β-adrenoceptors mediated NOS activation.

关 键 词：人脐静脉内皮细胞 Β-肾上腺素能受体 内皮型一氧化氮合酶 腺苷酸环化酶 蛋白激酶A 

分 类 号：R363[医药卫生—病理学]