Caveolae/caveolin-1/ERK 1/2 信号通路在降钙素基因相关肽抑制血管平滑肌细胞增殖中的作用（英文）
发布时间：2025-03-19 21:56
业已证明,Caveolae及其蛋白caveolin-1参与了细胞膜的胆固醇转运和细胞膜的信号转导.我们前期工作发现降钙素基因相关肽(CGRP)抑制血管平滑肌细胞(VSMC)增殖的信号通路与抑制ERK1/2活性和上调caveolin-1表达有关.本文研究Caveolae及caveolin-1在CGRP抑制VSMC增殖中的作用,进一步研究caveolin-1表达增加是否有直接抑制ERK1/2信号激酶活性的作用.采用大鼠主动脉贴块法培养VSMC,取3～10代VSMC用于实验,10%小牛血清(FBS)用于刺激VSMC增殖,用β-环糊精(cyclodextrin)或菲律宾菌素(filipin)剥夺胆固醇破坏Caveolae结构;MTT法和流式细胞仪用于检测细胞增殖;蛋白质印迹和免疫共沉淀法分别用于检测目的蛋白的表达或蛋白质间相互作用.结果显示,CGRP呈时间和浓度依赖性显著抑制10%FBS诱导的VSMC增殖.细胞Caveolae结构的破坏能降低CGRP抑制VSMC增殖作用,同时也增加了ERK1/2的磷酸化;β-环糊精孵育细胞能降低caveolin-1的表达.免疫共沉淀发现10%FBS和/或CGR...
【文章页数】：9 页
【文章目录】：
1 Materials and methods
1.1 Drugs and reagents
1.2 Rat vascular smooth muscle cell culture
1.3 Determination of cell proliferation
1.4 Western blotting
1.5 Immunoprecipitation
1.6 Statistical analysis
2 Results
2.1 Inhibitory effect of CGRP on VSMC proliferation
2.2 Effect ofβ-CD on the VSMC proliferation
2.3 Effect ofβ-CD on phosphorylation of ERK1/2
2.4 Effect of CGRP on expression of caveolin-1in the presence ofβ-CD
2.5 CGRP enhanced the interaction between caveolin-1 and p-ERK1/2
3 Discussion
本文编号：4036909
【文章页数】：9 页
【文章目录】：
1 Materials and methods
1.1 Drugs and reagents
1.2 Rat vascular smooth muscle cell culture
1.3 Determination of cell proliferation
1.4 Western blotting
1.5 Immunoprecipitation
1.6 Statistical analysis
2 Results
2.1 Inhibitory effect of CGRP on VSMC proliferation
2.2 Effect ofβ-CD on the VSMC proliferation
2.3 Effect ofβ-CD on phosphorylation of ERK1/2
2.4 Effect of CGRP on expression of caveolin-1in the presence ofβ-CD
2.5 CGRP enhanced the interaction between caveolin-1 and p-ERK1/2
3 Discussion
本文编号：4036909
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