降钙素基因相关肽抑制血管平滑肌细胞能量代谢的AMP激活蛋白激酶信号通路  

Signal Pathway of AMP-Activated Protein Kinase Involvement in the Inhibition of Energy Metabolism of Vascular Smooth Muscle Cell Induced by Calcitonin Gene-Related Peptide

作　　者：秦旭平[1] 任俊芳[1] 邓水秀[1] 吴峻[2] 谌赟[1] 陈临溪[1] 肖瑞平[3] 

机构地区：[1]南华大学药物药理研究所,湖南省衡阳市421001 [2]广州医学院附属第一医院,广东省广州市510120 [3]北京大学医学部心血管研究所,北京市100083

出　　处：《中国动脉硬化杂志》2008年第10期819-823,共5页Chinese Journal of Arteriosclerosis

基　　金：湖南省自然科学基金(05JJ30042);广东省自然科学基金(5300999)

摘　　要：目的观察降钙素基因相关肽对血管紧张素Ⅱ诱导血管平滑肌细胞增殖过程中能量代谢的影响及机制,探索细胞外信号调节激酶和AMP激活蛋白激酶相关信号蛋白在该作用中的地位。方法组织贴块法体外培养SD大鼠胸主动脉血管平滑肌细胞,取第3～10代用于实验。分别用降钙素基因相关肽或/和血管紧张素Ⅱ处理细胞。MTT法及细胞计数法观察降钙素基因相关肽对血管紧张素Ⅱ诱导大鼠血管平滑肌细胞增殖的影响;詹纳斯绿B染色观察细胞线粒体形态及体积;ATP检测试剂盒检测细胞内ATP水平;Western Blotting检测细胞磷酸化AMP激活蛋白激酶的表达。结果降钙素基因相关肽预处理能降低血管紧张素Ⅱ诱导的大鼠血管平滑肌细胞的增殖(P<0.05),同时拮抗血管紧张素Ⅱ引起细胞内线粒体肿胀、ATP水平升高,在此过程中伴随细胞内磷酸化AMP激活蛋白激酶表达下调(P<0.05);降钙素基因相关肽受体拮抗剂CGRP8-37能拮抗降钙素基因相关肽对血管紧张素Ⅱ促增殖和促代谢的抑制作用(P<0.05);细胞外信号调节激酶抑制剂PD98059能部分拮抗降钙素基因相关肽对磷酸化AMP激活蛋白激酶的抑制作用(P<0.05)。结论降钙素基因相关肽能显著抑制血管紧张素Ⅱ诱导大鼠血管平滑肌细胞增殖过程中的能量代谢,其细胞内信号通路可能涉及到降钙素基因相关肽/降钙素基因相关肽1型受体-丝裂原细胞外激酶/细胞外信号调节激酶-磷酸化AMP激活蛋白激酶信息传递链。Aim To investigate the effect of calcitonin gene-related peptide （ CGRP）on the proliferation and en- ergy metabolism of vascular smooth muscle cells （ VSMC ） induced by angiotensin Ⅱ （ Ang Ⅱ ） , and further explore whether AMP-activated protein kinase （AMPK） involved in the cellular signal pathway. Methods VSMC were prepared from thoracic aorta of male Sprague-Dawley rat by the explanted method. The 3 - 10 passages of ceils were used for the present studies. The viability of cultured VSMC was estimated by MTF assay and cells counting. Cellular energy metabolism was tested by ATP assay kit, Janus Green B staining. Western Blotting was used to observe the expressions of phospho- AMPK （p-AMPK） in VSMC. Results Ang 11 （100 nmol/L） increased the viability of VSMC. Pretreatment of the cell with CGRP significantly inhibited VSMC proliferation, hypertrophy of mitochondria, extra-generation of ATP induced by Ang Ⅱ （ P 〈 0.05 ）. Meanwhile, CGRP down-regulated expressions of p-AMPK induced by Ang Ⅱ , which were partly abolished by CGRPs_37 or PD98059, respectively. Conclusion Pretreatment with CGRP inhibits the proliferation and energy metabolism of VSMC induced by Ang Ⅱ. The signal transduction involving in this process is likely to be related with the signal linkage, namely, CGRP/CGRP receptor-1 （CGRPR-1 ） - mitogen extracellular kinase （ MEK）/ERK1/2- p- AMPK.

关 键 词：药理学 血管平滑肌细胞 降钙素基因相关肽 AMP激活蛋白激酶 能量代谢 

分 类 号：R96[医药卫生—药理学]