S-1-P 受体和人血管平滑肌细胞迁移在糖尿病和代谢综合征中的作用。
Role of S-1-P receptors and human vascular smooth muscle cell migration in diabetes and metabolic syndrome.
机构信息
Vascular Biology and Therapeutics Program, The Methodist Hospital Research Institute, Houston, TX, USA.
出版信息
J Surg Res. 2012 Oct;177(2):e75-82. doi: 10.1016/j.jss.2011.12.012. Epub 2012 Mar 11.
BACKGROUND
Sphingosine-1-phosphate (S-1-P) is a bioactive sphingolipid released from activated platelets that stimulates migration of vascular smooth muscle cells (VSMC) in vitro. S-1-P is associated with oxidized low-density lipoprotein (oxLDL) and is important in vessel remodeling. S-1-P will activate multiple G protein-coupled receptors (S-1-PR 1 to 5), which can regulate multiple cellular functions, including cell migration. The aim of this study is to examine the role of S-1-PR signaling during smooth muscle cell migration in response to S-1-P.
METHODS
Human VSMCs were cultured in vitro. Expression of S-1-PR 1 to 5 was determined in conditions mirroring diabetes (40 mM glucose) and metabolic syndrome (25 mM glucose with 20 μM linoleic acid and 20 μM oleic acid). Linear wound and Boyden microchemotaxis assays of migration were performed in the presence of S-1-P with and without siRNA against S-1-PR 1 to 5. Assays were performed for activation of ERK1/2, p38(MAPK) and JNK.
RESULTS
Human VSMCs express S-1-PR1, S-1-PR2, and S-1-PR3. There was no significant expression of S-1-PR4 and S-1-PR5. The expression of S-1-PR1 and S-1-PR3 is enhanced under high glucose conditions and metabolic syndrome conditions. Migration of VSMC in response to S-1-P is enhanced 2-fold by diabetes and 4-fold by metabolic syndrome. In diabetes, S-1-PR1 expression is enhanced, while S-1-PR2 and S-1-PR3 expression are both maintained. In metabolic syndrome, S-1-PR1 and 3 expressions are enhanced and that of S-1-PR2 is reduced. siRNA to S-1-PR1 results in a 2-fold reduction in S-1-P-mediated cell migration under all conditions. siRNA to S-1-PR2 enhanced cell migration only under normal conditions, while siRNA S-1-PR3 decreased migration in metabolic syndrome only. Down-regulation of S-1-PR1 reduced ERK1/2 activation in response to S-1-P, while that of S-1-PR2 had no effect under normal conditions. In diabetes, down-regulation of S-1-PR1 reduced activation of all three MAPKs. In metabolic syndrome, down-regulation of S-1-PR1 and S-1-PR3 reduced activation of all three MAPKs.
CONCLUSION
S-1-PR 1, 2, and 3 regulate human VSMC migration and their expression level and function are modulated by conditions simulating diabetes and metabolic syndrome.
背景
鞘氨醇-1-磷酸（S-1-P）是一种从激活的血小板中释放的生物活性鞘脂，可刺激体外血管平滑肌细胞（VSMC）的迁移。S-1-P 与氧化型低密度脂蛋白（oxLDL）有关，在血管重塑中很重要。S-1-P 将激活多种 G 蛋白偶联受体（S-1-PR1 至 5），这些受体可以调节多种细胞功能，包括细胞迁移。本研究旨在探讨 S-1-PR 信号在 S-1-P 诱导的平滑肌细胞迁移中的作用。
方法
在模拟糖尿病（40mM 葡萄糖）和代谢综合征（25mM 葡萄糖加 20μM 亚油酸和 20μM 油酸）的条件下，培养人 VSMC。测定 S-1-PR1 至 5 的表达。在存在 S-1-P 以及针对 S-1-PR1 至 5 的 siRNA 的情况下，进行线性划痕和 Boyden 微趋化运动测定。测定 ERK1/2、p38（MAPK）和 JNK 的激活。
结果
人 VSMC 表达 S-1-PR1、S-1-PR2 和 S-1-PR3。S-1-PR4 和 S-1-PR5 的表达不显著。S-1-PR1 和 S-1-PR3 的表达在高糖条件和代谢综合征条件下增强。S-1-P 诱导的 VSMC 迁移在糖尿病中增强 2 倍，在代谢综合征中增强 4 倍。在糖尿病中，S-1-PR1 的表达增强，而 S-1-PR2 和 S-1-PR3 的表达均保持不变。在代谢综合征中，S-1-PR1 和 S-1-PR3 的表达增强，S-1-PR2 的表达降低。在所有条件下，S-1-PR1 的 siRNA 使 S-1-P 介导的细胞迁移减少 2 倍。S-1-PR2 的 siRNA 仅在正常条件下增强细胞迁移，而 S-1-PR3 的 siRNA 在代谢综合征中仅降低迁移。下调 S-1-PR1 可减少 S-1-P 诱导的 ERK1/2 激活，而 S-1-PR2 在正常条件下无影响。在糖尿病中，下调 S-1-PR1 可降低所有三种 MAPK 的激活。在代谢综合征中，下调 S-1-PR1 和 S-1-PR3 可降低所有三种 MAPK 的激活。
结论
S-1-PR1、2 和 3 调节人 VSMC 迁移，其表达水平和功能受模拟糖尿病和代谢综合征的条件调节。
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