Matthews L, Berry A, Tersigni M, D'Acquisto F, Ianaro A, Ray D
Endocrine Sciences Research Group, Division of Cardiovascular and Endocrine Science, University of Manchester, Manchester M13 9PT, United Kingdom.
Endocrinology. 2009 Jan;150(1):75-86. doi: 10.1210/en.2008-0196. Epub 2008 Sep 18.
Although thiazolidinediones were designed as specific peroxisome proliferator-activated receptor (PPAR)-gamma-ligands, there is evidence for some off-target effects mediated by a non-PPARgamma mechanism. Previously we have shown that rosiglitazone has antiinflammatory actions not explicable by activation of PPARgamma,but possibly by the glucocorticoid receptor (GR). Rosiglitazone induces nuclear translocation both of GR-green fluorescent protein, and endogenous GR in HeLa and U20S cells but with slower kinetics than dexamethasone. Rosiglitazone also induces GR phosphorylation (Ser211), a GR ligand-binding-specific effect. Rosiglitazone drives luciferase expression from a simple glucocorticoid-response element containing reporter gene in a GR-dependent manner (EC50 4 microm), with a similar amplitude response to the partial GR agonist RU486. Rosiglitazone also inhibits dexamethasone-driven reporter gene activity (IC50 2.9 microm) in a similar fashion to RU486, suggesting partial agonist activity. Importantly we demonstrate a similar effect in PPARgamma-null cells, suggesting both GR dependence and PPARgamma independence. Rosiglitazone also activates a GAL4-GR chimera, driving a upstream activating sequence promoter, demonstrating DNA template sequence independence and furthermore enhanced steroid receptor coactivator-1-GR interaction, measured by a mammalian two-hybrid assay. Both ciglitazone and pioglitazone, structurally related to rosiglitazone, show similar effects on the GR. The antiproliferative effect of rosiglitazone is increased in U20S cells that overexpress GR, suggesting a biologically important GR-dependent component of rosiglitazone action. Rosiglitazone is a partial GR agonist, affecting GR activation and trafficking to influence engagement of target genes and affect cell function. This novel mode of action may explain some off-target effects observed in vivo. Additionally, antagonism of glucocorticoid action may contribute to the antidiabetic actions of rosiglitazone.
尽管噻唑烷二酮类药物被设计为特异性过氧化物酶体增殖物激活受体(PPAR)-γ配体,但有证据表明存在一些由非PPARγ机制介导的脱靶效应。此前我们已表明,罗格列酮具有抗炎作用,这无法通过PPARγ的激活来解释,而可能是通过糖皮质激素受体(GR)介导的。罗格列酮可诱导GR-绿色荧光蛋白以及HeLa和U20S细胞中的内源性GR发生核转位,但动力学比地塞米松慢。罗格列酮还可诱导GR磷酸化(Ser211),这是一种GR配体结合特异性效应。罗格列酮以GR依赖的方式(EC50为4微摩尔)驱动来自含简单糖皮质激素反应元件的报告基因的荧光素酶表达,对部分GR激动剂RU486的反应幅度相似。罗格列酮还以与RU486类似的方式抑制地塞米松驱动的报告基因活性(IC50为2.9微摩尔),表明具有部分激动剂活性。重要的是,我们在PPARγ缺失的细胞中证明了类似的效应,表明既依赖GR又不依赖PPARγ。罗格列酮还可激活GAL4-GR嵌合体,驱动上游激活序列启动子,证明与DNA模板序列无关,此外,通过哺乳动物双杂交试验检测发现其增强了类固醇受体共激活因子-1与GR的相互作用。与罗格列酮结构相关的环格列酮和吡格列酮对GR显示出类似的作用。在过表达GR的U20S细胞中,罗格列酮的抗增殖作用增强,这表明罗格列酮作用中存在生物学上重要的GR依赖成分。罗格列酮是一种部分GR激动剂,影响GR的激活和转运,从而影响靶基因的结合并影响细胞功能。这种新的作用模式可能解释了体内观察到的一些脱靶效应。此外,糖皮质激素作用的拮抗可能有助于罗格列酮的抗糖尿病作用。