Inhibitory Effect of Idelalisib, a Selective Phosphatidylinositol 3-Kinase δ Inhibitor, on Adipogenesis in an In Vitro Model of Graves' Orbitopathy
Inhibitory Effect of Idelalisib, a Selective Phosphatidylinositol 3-Kinase δ Inhibitor, on Adipogenesis in an In Vitro Model of Graves' Orbitopathy
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DOI：
10.1167/iovs.18-24509
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发表时间：
2018-09-01
影响因子：
4.4
通讯作者：
Yoon, Jin Sook
中科院分区：
文献类型：
--
作者：
Ko, JaeSang;Kim, Ji-Young;Yoon, Jin Sook
目的：新的证据表明磷脂酰肌醇3-激酶(PI3K)-AKT通路参与了Graves眼眶病(GO)的发病过程。本研究观察选择性PI3K Delta抑制剂idelalisib对体外培养的GO眼眶成纤维细胞的成脂作用。在IL-1β刺激细胞前，先用艾德拉西布预处理1h。免疫印迹和ELISAs法检测炎性细胞因子的表达。用Western blotting方法检测PI3K/AKT信号通路中与脂肪形成相关的下游分子叉头盒蛋白O1(FOXO1)和哺乳动物靶标雷帕霉素(MTOR)的表达。成脂分化和艾达利西处理后，用油红O染色，Western印迹法检测细胞中PPAR-γ和CCAAT-增强子结合蛋白(C/EBP)α/β的表达水平。结果：艾得拉西布处理GO和非GO眼眶成纤维细胞后，AKT磷酸化水平呈剂量依赖性降低。Idelalisib可抑制IL-1β诱导的IL-6和IL-8的表达。Idelalisib减弱了PI3K途径下游调节因子mTOR和FOXO1的磷酸化。油红-O染色结果显示脂滴减少，PPAR？结论：idelalisib可抑制GO眼眶成纤维细胞的促炎细胞因子产生和成脂作用。这些结果支持了PI3K抑制剂在GO管理中的潜在应用。
Purpose: Emerging evidence indicates that the phosphatidylinositol 3-kinase (PI3K)-AKT pathway is involved in the pathogenesis of Graves' orbitopathy (GO). In this study, the therapeutic effects of idelalisib, a selective PI3K delta inhibitor, on adipogenesis were evaluated in GO orbital fibroblasts in vitro.Methods: Orbital fibroblasts were cultured from orbital connective tissues obtained from individuals with GO and healthy control subjects. Cells were pretreated with idelalisib for 1 hour before stimulation with IL-1 beta. Inflammatory cytokine expression was measured by Western blotting and ELISAs. The adipogenesis-related downstream mediators of the PI3K/AKT cascade, that is, forkhead box protein O1 (FOXO1) and mammalian target of rapamycin (mTOR), also were measured by Western blotting. After adipogenic differentiation and idelalisib treatment, cells were stained with Oil Red O and the levels of peroxisome proliferator activator gamma (PPAR gamma) and CCAAT-enhancer-binding proteins (C/EBP) alpha/beta were determined by Western blot analyses.Results: AKT phosphorylation decreased in a dose-dependent manner upon treatment with idelalisib in GO and non-GO orbital fibroblasts. Treatment with idelalisib inhibited the IL-1 beta-induced expression of IL-6 and IL-8. Idelalisib attenuated the phosphorylation of mTOR and FOXO1, downstream regulators of the PI3K pathway. Oil Red-O staining results revealed a decrease in lipid droplets and suppressed expression of PPAR? and c/EBP alpha/beta upon treatment with idelalisib during adipose differentiation.Conclusions: Idelalisib inhibited proinflammatory cytokine production and adipogenesis in GO orbital fibroblasts in vitro. These results support the potential use of PI3K inhibitors in GO management.