KCa3.1 channels mediate the increase of cell migration and proliferation by advanced glycation endproducts in cultured rat vascular smooth muscle cells
KCa3.1 channels mediate the increase of cell migration and proliferation by advanced glycation endproducts in cultured rat vascular smooth muscle cells
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DOI：
10.1038/labinvest.2012.163
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发表时间：
2013-02-01
影响因子：
5
通讯作者：
Deng, Xiu-Ling
中科院分区：
文献类型：
--
作者：
Zhao, Li-Mei;Su, Xing-Li;Deng, Xiu-Ling
晚期糖基化终产物（AGEs）参与糖尿病动脉粥样硬化的机制尚未完全清楚。本研究采用全细胞膜片钳技术、细胞增殖和迁移实验以及蛋白质印迹分析等方法，探讨AGEs诱导的大鼠血管平滑肌细胞（VSMC）迁移和增殖是否与中电导钙激活的钾通道（K（Ca）3.1通道）有关。结果发现，AGE-BSA（牛血清白蛋白）可使细胞K（Ca）3.1通道电流密度和蛋白水平增加，而AGE-BSA与抗AGEs受体抗体（anti-receptors of AGEs）共孵育可逆转这种作用。ERK 1/2抑制剂PD 98059和U 0126、P38-MAPK抑制剂SB 203580和SB 202190、PI 3 K抑制剂LY 294002和wortmannin可抑制AGE-BSA诱导的K（Ca）3.1通道表达。此外，AGE-BAS增加了细胞迁移和增殖，并且这种作用可以用抗钙离子抗体、K（Ca）3.1通道阻断剂TRAM-34或K（Ca）3.1小干扰RNA完全逆转。这些结果首次证明AGEs诱导的大鼠VMSC迁移和增殖的增加与K（Ca）3.1通道的上调有关，并且细胞内信号ERK 1/2、P38-MAPK和PI 3 K参与了K（Ca）3.1通道表达的调节。实验室调查（2013）93，159-167; doi：10.1038/labinvest.2012.163; 2012年11月19日在线发表
The mechanisms underlying the involvement of advanced glycation endproducts (AGEs) in diabetic atherosclerosis are not fully understood. The present study was designed to investigate whether intermediate-conductance Ca2+-activated K+ channels (K(Ca)3.1 channels) are involved in migration and proliferation induced by AGEs in cultured rat vascular smooth muscle cells (VSMCs) using approaches of whole-cell patch voltage:clamp, cell proliferation and migration assay, and western blot analysis. It was found that the current density and protein level of K(Ca)3.1 channels were enhanced in cells incubated with AGE-BSA (bovine serum albumin), and the effects were reversed by co-incubation of AGE-BSA with anti-RAGE (anti-receptors of AGEs) antibody. The ERK1/2 inhibitors PD98059 and U0126, the P38-MAPK inhibitors SB203580 and SB202190, or the PI3K inhibitors LY294002 and wortmannin countered the K(Ca)3.1 channel expression by AGE-BSA. In addition, AGE-BAS increased cell migration and proliferation, and the effects were fully reversed with anti-RAGE antibody, the K(Ca)3.1 channel blocker TRAM-34, or K(Ca)3.1 small interfering RNA. These results demonstrate for the first time that AGEs-induced increase of migration and proliferation is related to the upregulation of K(Ca)3.1 channels in rat VMSCs, and the intracellular signals ERK1/2, P38-MAPK and PI3K are involved in the regulation of K(Ca)3.1 channel expression. Laboratory Investigation (2013) 93, 159-167; doi:10.1038/labinvest.2012.163; published online 19 November 2012