PI3K—Akt—eNOS信号转导通路在吗啡促人脐静脉内皮细胞一氧化氮合成中的作用  

Role of PDK-Akt-eNOS signal transduction pathway in nitric oxide synthesis increased by morphine in human umbilical vein endothefial cells

作　　者：王志[1] 赵惠娟[2] 王飞[1] 纪风涛[1] 车月娟[1] 李玉娟[1] 彭书峻[1] 

机构地区：[1]中山大学附属第二医院麻醉科,广州市510120 [2]中山大学附属第一医院手术室,广州市510120

出　　处：《中华麻醉学杂志》2009年第8期737-740,共4页Chinese Journal of Anesthesiology

基　　金：广东省医学科研基金（A2008179）

摘　　要：目的评价磷脂酰肌醇-3激酶-蛋白质丝氨酸苏氨酸激酶-内皮型一氧化氮合酶（PI3K-Akt—eNOS）信号转导通路在吗啡对促人脐静脉内皮细胞一氧化氮（NO）合成中的作用。方法体外培养人脐静脉内皮细胞，实验Ⅰ：人脐静脉内皮细胞随机分为4组，每组6孔：C组和M1-2组，C组不予任何处理；M1-3组加入吗啡，终浓度为1μmol／L，分别孵育3、6和12h，随后测定NO含量；实验Ⅱ：人脐静脉内皮细胞随机分为4组，每组6孔：C组和M1-3组，C组不予任何处理；M1-3组加入吗啡，终浓度分别为0．01、1和100μmol／L，孵育6h，随后测定NO含量；实验Ⅲ：人脐静脉内皮细胞随机分为5组，每组6孔：C组、M组、MW组、MS组和ML组，C组不予任何处理；M组加入吗啡，终浓度1μmol／L；MW组先加入wortmannin（P13K特异性抑制剂），15min后加入吗啡，终浓度分别为5、1μmol／L；MS组先加入SH-5（Akt特异性抑制剂），15min后加入吗啡，终浓度分别为10、1μmol／L；ML组先加入L-NAME（eNOS特异性抑制剂），15min后加入吗啡，终浓度分别为0．5、1μmol／L，各组孵育6h后，测定NO含量。结果实验Ⅰ结果：随吗啡孵育时间延长，促脐静脉内皮细胞NO合成的作用增强（P〈0．05）；实验Ⅱ结果：随吗啡浓度升高，促脐静脉内皮细胞NO合成的作用增强（P〈0．05）；实验Ⅲ结果：与C组比较，M组人脐静脉内皮细胞NO含量升高（P〈0．05），MW组、MS组和ML组差异无统计学意义（P〉0．05）；与M组比较，MW组、MS组和ML组人脐静脉内皮细胞NO含量降低（P〈0．05）。结论吗啡促人脐静脉内皮细胞NO合成呈浓度和时间依赖性，其机制可能与激活PI3K-Akt-eNOS信号转导通路有关。Objective To evaluate the role of phosphatidylinositol 3-kinase-Akt-emlothelial nitric oxide synthase （PI3K/Akt/cNOS） signal transduction pathway in nitric oxide （NO） synthesis increased by morphine in cultured human umbilical vein endothelial cells （HUVECs）. Methods HUVECs were inoculated in 24 well palates in vitro. Three experiments were performed in this study. In experiment Ⅰ , HUVECs were randomly divided into 4 groups of 6 wells each： control group （group C） and morphine group （group M1-3） . In group M1-3 , morphine with the final concentration of 1 μmol/L was added, and then the cells were incubated for 3, 6 and 12 h respectively. NO synthesis was viewed with fluorescence microscope and NO content was measured. In experiment Ⅱ , HUVECs were randomly divided into 4 groups of 6 wells each： group C and group M1-3 . In group M1-3 , morphine with the final concentrations of 0.01, 1 and 100 μmol/L was added respectively, and then the cells were incubated for 6 h. NO content was measured. In experiment Ⅲ , HUVECs were randomly divided into 5 groups of 6 wells each： group C, group M, group M ＋ wortmannin （W） （group MW）, group M ＋ SH-5 （group MS） and group M ＋ L-NAME （group ML）. In group M, morphine with the final concentration of 1 μmol/L was added. In group MW, wortmannin （a PI3K specific inhibitor） was added and 15 min later, morphine with the final concentrations of 5 and 1 μmol/L was added. In group MS, SH-5 （an Akt specific inhibitor） was added and 15 min later, morphine with the final concentrations of 10 and 1 μmol/L was added. In group ML, L-NAME （an eNOS specific inhibitor） was added and 15 min later, morphine with the final concentrations of 0.5 and 1 μmol/L was added. The cells were then incubated for 6 h and NO content was measured in each group. Results In experiment Ⅰ , morphine treatment increased the endothelial NO synthesis in a time-dependent manner （P 〈 0.05）. In experiment Ⅱ , morphine treatment increased the

关 键 词：1-磷脂酰肌醇3-激酶 蛋白质丝氨酸苏氨酸激酶 一氧化氮合酶 吗啡 内皮细胞 脐静脉 一氧化氮 

分 类 号：R285.5[医药卫生—中药学] R977.12[医药卫生—中医学]