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膜雌激素受体介导一氧化氮合酶活性增高的快速非基因效应

王庭槐, 付晓东, 杨丹, 谈智, 潘敬运

中山大学医学院生理学教研室，广州510080

摘要

实验利用新生小牛胸主动脉内皮细胞(BAECs)作为模型，观察1 7 [r雌二醇(E2) , E,BSA对BAEC、中内皮型一氧化氮合酶(e NOS)的快速激活作用，并探讨了丝裂素活化蛋白激酶( MAP K)信号通路在其中的作用。结果显示，不同浓度的E2(0. 001一1 umol/ L)作用于BAECs 15 min均能快速激活eNOS; 0.01 umol/L浓度的E2作用于BAECs, 5 min即能激活e NOS , 1 5 min达到最大效应，随后e N OS快速失活;E2BSA (17. 5 ng/ ml)作用于BAECs, 1 5min同样可激活eNOSo E2. E2BSA激活e NOS的作用均能被雌激素受体(ER)拮抗剂tamoxifen (0-1 umol/ L)或MAP K激酶特异抑制剂PD98059 (50 11 mol/ L)所阻断。放线菌素。(25 ug/ ml)不能阻断E2、E2BSA对e N OS的激活作用。E2(0. 01 umol/ L) , E2BSA (17.5 ng/ ml)作用于BAECs 15 min后可明显促进p42/ p44磷酸化MAPK蛋白表
达，而对p42/p44 MAPK总蛋白表达无影响。Ta m oxife n可部分阻断E2 . E2BSA激活p42/ p44磷酸化MAPK的作用。这些结果提示，BAECs膜上可能存在膜雌激素受体(membrane estrogen receptor, mER)，E2 . E2BSA作用于mER后可通过MAPK信号途径快速激活e NOS .

关键词： 雌激素; 膜雌激素受体; 一氧化氮合酶; 丝裂素活化蛋白激酶; 血管内皮细胞

Membrane estrogen receptor mediates the rapid nongenomic activation of endothelial nitric oxide synthase by estrogen

WANG Ting Huai, FU Xiao- Dong, Yang Dan, Tan Zhi, PAN Jing Yun

Department of Physiology，Sun Yat-sen University of Medical Sciences，Guangzhou 510080

Abstract

In the present study，confluent bovine aortic endothelial cells(BAECs) were used to study the rapid nongenomic effects of 17f}-estradiol and the membrane impermeable conjugated 17f}-estradiol (EZBSA) on the activation of endothelial nitric oxide synthase(eNOS) and mitogen activated protein kinase(MAP K) .eNOS activation was assessed in whole cells by measuring[3H]L-arginine conversion to [3H]L- citrulline .MAPK activity was determined by Western Slotting .The results obtained show that the addition of various concentrations of EZ(0. 001一1 p mol/ L) resulted in 122士29，186士17，83士20
and 157士29%increases in eNOS activity，respectively，in BAECs within 15 min of exposure to the hor mone .E2(0-01 p mol/ L)一stimulated eNOS activity was detectable during 5-，15- and 30- min incubation which yielded increases of 37士6，56士9 and 38士8%，respectively .The increase reached a plateau from
15 through 30 min and rapidly declined thereafter.EZ BS A(17. 5 ng/ ml) also enhanced eNOS activity by an increase of 35士9%above the basal activity .The effect of EZ and EZBSA on eNOS activation was unaffected by actinomycin D(25 pg/ ml) but was obviously inhibited by tamoxifen(0.1 p mol/ L) and
PD98059(50 p mol/ L).Compared with control，EZ and EZBSA sti mulation of BAECs for 1 5 min caused an increase in MAP K activity by 428士17 and 360士14%respectively.This effect was blocked by tamoxifen.These results suggest that there might be the me mbrane estrogen receptor localized on BAECs，
which mediates the rapid nongenomic effect of estrogen on eNOS activation through MAPK pathways.

Key words: Estrogen;me mbrane estrogen receptor;nitric oxide synthase;mitogen activated protein kinase;vascular endothelial cell

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引用本文：

王庭槐, 付晓东, 杨丹, 谈智, 潘敬运. 膜雌激素受体介导一氧化氮合酶活性增高的快速非基因效应[J]. 生理学报 2003; 55 (2): .

WANG Ting Huai, FU Xiao- Dong, Yang Dan, Tan Zhi, PAN Jing Yun. Membrane estrogen receptor mediates the rapid nongenomic activation of endothelial nitric oxide synthase by estrogen. Acta Physiol Sin 2003; 55 (2): (in Chinese with English abstract).