Shimamura Ken, Takashiro Yuko, Akiyama Nobuteru, Hirabayashi Tetsuya, Murayama Toshihiko
Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 263-8522, Japan.
Eur J Pharmacol. 2004 Feb 20;486(2):141-50. doi: 10.1016/j.ejphar.2003.12.022.
We investigated the effects of sphingosine 1-phosphate and histamine on the expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and E-selectin, and their signaling pathways in human umbilical vein endothelial cells. Sphingosine 1-phosphate increased the mRNA and protein level of VCAM-1, and the mRNAs of E-selectin and ICAM-1. The effects of sphingosine 1-phosphate were inhibited by the pertussis toxin and the respective inhibitors (10 microM 1-[6-[[(17beta)-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione (U73122) for phosphoinositide-specific phospholipase C; 10 microM 4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-1H-imidazole (SB203580) for p38 mitogen-activated protein kinase (MAPK); 1 microM 12-(2-cyanoethyl)-6,7,12,13-tetrahydro-13-methyl-5-oxo-5H-indolo(2,3-a)pyrrolo(3,4-c)-carbazole (Gö6976) for the alpha form of protein kinase C (PKC-alpha)), but not by a PKC-delta inhibitor (1 microM rottlerin). Histamine, which alone showed no effect, enhanced the sphingosine 1-phosphate-induced expressions via histamine H(1) receptor. The histamine response decreased by U73122 and rottlerin, but not by SB203580 and Gö6976. The effects of sphingosine 1-phosphate with and without histamine were abolished by the higher concentrations of PKC inhibitors and in the PKC-depleted cells. Sphingosine 1-phosphate and histamine alone stimulated phosphorylation of p38 MAPK in a phosphoinositide-specific phospholipase C-dependent but not in a PKCs-independent manner. These findings suggest that sphingosine 1-phosphate-induced expression of adhesion molecules was mediated by phosphoinositide-specific phospholipase C and preferentially by PKC-alpha and p38 MAPK, and the histamine response was mediated by phosphoinositide-specific phospholipase C and PKC-delta in human umbilical vein endothelial cells.
我们研究了1-磷酸鞘氨醇和组胺对人脐静脉内皮细胞中血管细胞黏附分子-1（VCAM-1）、细胞间黏附分子-1（ICAM-1）和E-选择素表达及其信号通路的影响。1-磷酸鞘氨醇增加了VCAM-1的mRNA和蛋白水平，以及E-选择素和ICAM-1的mRNA水平。百日咳毒素和相应抑制剂（针对磷脂酰肌醇特异性磷脂酶C的10 microM 1-[6-[[(17β)-3-甲氧基雌甾-1,3,5(10)-三烯-17-基]氨基]己基]-1H-吡咯-2,5-二酮（U73122）；针对p38丝裂原活化蛋白激酶（MAPK）的10 microM 4-(4-氟苯基)-2-(4-甲基亚磺酰基苯基)-5-(4-吡啶基)-1H-咪唑（SB203580）；针对蛋白激酶Cα（PKC-α）的1 microM 12-(2-氰基乙基)-6,7,12,13-四氢-13-甲基-5-氧代-5H-吲哚并(2,3-a)吡咯并(3,4-c)咔唑（Gö6976））可抑制1-磷酸鞘氨醇的作用，但PKC-δ抑制剂（1 microM罗特勒素）则无此作用。单独作用时无效应的组胺，通过组胺H(1)受体增强了1-磷酸鞘氨醇诱导的表达。U73122和罗特勒素可降低组胺反应，但SB203580和Gö6976则无此作用。较高浓度的PKC抑制剂以及PKC缺失的细胞可消除1-磷酸鞘氨醇单独作用和与组胺共同作用时的效应。1-磷酸鞘氨醇和组胺单独作用时，以磷脂酰肌醇特异性磷脂酶C依赖但非PKC非依赖的方式刺激p38 MAPK的磷酸化。这些发现表明，1-磷酸鞘氨醇诱导的黏附分子表达由磷脂酰肌醇特异性磷脂酶C介导，且优先由PKC-α和p38 MAPK介导，而组胺反应在人脐静脉内皮细胞中由磷脂酰肌醇特异性磷脂酶C和PKC-δ介导。
