摘要
目的观察肾上腺髓质素(ADM)对兔血管内皮细胞内信号转导途径的影响。方法用原代培养的兔血管内皮细胞,给予肾上腺髓质素、cAMP激动剂forskolin(10-7mol/L)、NO供体L-精氨酸(1 mmol/L)、NO合酶抑制剂L-NMMA(10-5mol/L)、异丙肾上腺素(10-6mol/L)、8-bromo-AMP(10-7mol/L)及cGMP抑制剂L-NAME(10-4mol/L),观察细胞内cAMP、cGMP水平及Ca2+荧光强度的变化。用酶联免疫法测定cAMP及cGMP。用Fluo-3AM荧光负载兔血管内皮细胞测定细胞内钙浓度。结果肾上腺髓质素呈浓度依赖性升高兔血管内皮细胞内cAMP,而降低cGMP水平。肾上腺髓质素可使钙离子(Ca2+)水平下降(P<0.001),L-NMMA预处理组无明显作用,当加入L-精氨酸后荧光强度急剧下降。结论肾上腺髓质素能够影响兔血管内皮细胞的cAMP、cGMP及细胞内钙水平,从而实现其对细胞的多种生物学效应。
Objctive To study the signal transduction pathways of adrenomedullin on primary cultured rabbit vascular endothelial cells. Methods After the adrenomedullin, forskolin （ 10^-7 mol/L） and 8-bromo-AMP （ 10^-7 mol/L） which both can increase the cAMP level of cells, control group L-NAME （10^-4 mol/L）can decrease the cGMP level of cells were added into cells for 60 minutes, the level of cAMP and cGMP were measured by immunoassay KIT. The concentration of intracellular calcium was examed by the fluo-3 probe. Four group of adrenomedullin（ 10^-7-10^-10 mol/L）, L-arginine （lmmol/L） , L-NMMA（inhibitor of of nitro oxidease, 10^-5mol/L ） and isoprenaline （10^-3mol/L） were added to the vascular endothelial ceils with fluo-3 in order to determine the changes of fluorescence. Results The effects of adrenomedullin for increasing cAMP on the vascular endothelial cells showed concentration independence. The positive control forskolin and 8-bromo-AMP didn＇t have this action. The effects of adrenomedullin for decreasing cGMP on the vascular endothelial cells showed concentration independence as well. The four kinds of adrenomedullin all could reduce the con-centration of fluorescence compared with controls （ P 〈 0.01 ）. The concentration of fluorescence was decreased obvi-ously with L-arginine to the minimal value. Adrenomedullin affected signal transduction of vascular endothelial cells by influcing cAMP.cGMP and the calcium of mobilizaion. Conclusion With the above actions adrenomedullin can exert many biological actions on endothelial vascular cells.
出处
《基础医学与临床》
CSCD
北大核心
2005年第11期1020-1023,共4页
Basic and Clinical Medicine
基金
国家自然科学基金(39970780)
