Sun Haipeng, Xu Beibei, Sheveleva Elena, Chen Qin M
Interdisciplinary Graduate Program of Pharmacology and Toxicology, University of Arizona, Tucson, AZ 85724, USA.
Toxicol Appl Pharmacol. 2008 Oct 1;232(1):25-32. doi: 10.1016/j.taap.2008.05.024. Epub 2008 Jun 4.
Glucocorticoids induce COX-2 expression in rat cardiomyocytes. While investigating whether phosphatidylinositol 3 kinase (PI3K) plays a role in corticosterone (CT)-induced COX-2, we found that LY294002 (LY29) but not wortmannin (WM) attenuates CT from inducing COX-2 gene expression. Expression of a dominant-negative mutant of p85 subunit of PI3K failed to inhibit CT from inducing COX-2 expression. CT did not activate PI3K/AKT signaling pathway whereas LY29 and WM decreased the activity of PI3K. LY303511 (LY30), a structural analogue and a negative control for PI3K inhibitory activity of LY29, also suppressed COX-2 induction. These data suggest PI3K-independent mechanisms in regulating CT-induced COX-2 expression. LY29 and LY30 do not inhibit glucocorticoid receptor transactivity. Both compounds have been reported to inhibit Casein Kinase 2 activity and modulate potassium and calcium levels independent of PI3K, while LY29 has been reported to inhibit mammalian Target of Rapamycin (mTOR), and DNA-dependent Protein Kinase (DNA-PK). Inhibitor of Casein Kinase 2 (CK2), mTOR or DNA-PK failed to prevent CT from inducing COX-2 expression. Tetraethylammonium (TEA), a potassium channel blocker, and nimodipine, a calcium channel blocker, both attenuated CT from inducing COX-2 gene expression. CT was found to increase intracellular Ca(2+) concentration, which can be inhibited by LY29, TEA or nimodipine. These data suggest a possible role of calcium instead of PI3K in CT-induced COX-2 expression in cardiomyocytes.
糖皮质激素可诱导大鼠心肌细胞中COX - 2的表达。在研究磷脂酰肌醇3激酶(PI3K)是否在皮质酮(CT)诱导COX - 2的过程中发挥作用时,我们发现LY294002(LY29)而非渥曼青霉素(WM)可减弱CT诱导的COX - 2基因表达。PI3K的p85亚基的显性负性突变体的表达未能抑制CT诱导的COX - 2表达。CT未激活PI3K/AKT信号通路,而LY29和WM降低了PI3K的活性。LY303511(LY30),一种结构类似物且是LY29的PI3K抑制活性的阴性对照,也抑制了COX - 2的诱导。这些数据表明在调节CT诱导的COX - 2表达中存在不依赖PI3K的机制。LY29和LY30不抑制糖皮质激素受体的反式激活。据报道这两种化合物均可抑制酪蛋白激酶2的活性并独立于PI3K调节钾和钙水平,而据报道LY29可抑制哺乳动物雷帕霉素靶蛋白(mTOR)和DNA依赖性蛋白激酶(DNA - PK)。酪蛋白激酶2(CK2)、mTOR或DNA - PK的抑制剂未能阻止CT诱导COX - 2表达。钾通道阻滞剂四乙铵(TEA)和钙通道阻滞剂尼莫地平均减弱了CT诱导的COX - 2基因表达。发现CT可增加细胞内Ca(2+)浓度,而这可被LY29、TEA或尼莫地平抑制。这些数据表明在心肌细胞中,钙而非PI3K在CT诱导的COX - 2表达中可能发挥作用。