State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Acta Pharmacol Sin. 2011 Sep;32(9):1100-8. doi: 10.1038/aps.2011.69. Epub 2011 Aug 1.
To examine the neuroprotective effects of T33, a peroxisome proliferator-activated receptor gamma/alpha (PPARγ/α) agonist, in acute ischemic models in vitro and in vivo.
Primary astrocytes subjected to oxygen-glucose deprivation/reperfusion (O/R) and BV-2 cells subjected to hypoxia were used as a model simulating the ischemic core and penumbra, respectively. The mRNA levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured using qPCR. The levels of TNF-α secreted by BV-2 cells were measured using ELISA. Protein levels of cyclooxygenase-2 (COX-2), p65, phosphorylated I-κBα/I-κBα, phosphorylated I-κB kinase (pIKK), phosphorylated eukaryote initiation factor 2α (p-eIF-2α)/eIF-2α and p-p38/p38 were detected using Western blot. PPARγ activity was measured using EMSA. The neuroprotection in vivo was examined in rat middle cerebral artery occlusion (MCAO) model with neurological scoring and TTC staining.
Addition of T33 (0.5 μmol/L) increased the level of I-κBα protein in primary astrocytes subjected to O/R, which was due to promoting protein synthesis without affecting degradation. In primary astrocytes subjected to O/R, addition of T33 amplified I-κBα gene transcription and mRNA translation, thus suppressing the nuclear factor-kappa B (NF-κB) pathway and reducing inflammatory mediators (TNF-α, IL-1β, and COX-2). In BV-2 cells subjected to hypoxia, T33 (0.5 μmol/L) reduced TNF-α, COX-2, and p-P38 production, which was antagonized by pre-administration of the specific PPARγ antagonist GW9662 (30 μmol/L). T33 (2 mg/kg, ip) attenuated MCAO-induced inflammatory responses and brain infarction, which was antagonized by pre-administered GW9662 (4 mg/kg, ip).
T33 exerted anti-inflammatory effects in the ischemic core and penumbra via PPARγ activation, which contributed to its neuroprotective action.
研究过氧化物酶体增殖物激活受体γ/α（PPARγ/α）激动剂 T33 在体外和体内急性缺血模型中的神经保护作用。
采用氧葡萄糖剥夺/再灌注（O/R）的原代星形胶质细胞和缺氧的 BV-2 细胞作为模拟缺血核心和半影区的模型。使用 qPCR 测量肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）的 mRNA 水平。使用 ELISA 测量 BV-2 细胞分泌的 TNF-α 水平。使用 Western blot 检测环氧化酶-2（COX-2）、p65、磷酸化 I-κBα/I-κBα、磷酸化 I-κB 激酶（pIKK）、磷酸化真核起始因子 2α（p-eIF-2α）/eIF-2α 和 p-p38/p38 的蛋白水平。使用 EMSA 测量 PPARγ 活性。通过神经功能评分和 TTC 染色检测大鼠大脑中动脉闭塞（MCAO）模型中的体内神经保护作用。
添加 T33（0.5μmol/L）增加了 O/R 处理的原代星形胶质细胞中 I-κBα 蛋白的水平，这是由于促进蛋白合成而不影响降解所致。在 O/R 处理的原代星形胶质细胞中，T33 增强了 I-κBα 基因转录和 mRNA 翻译，从而抑制核因子-κB（NF-κB）通路并减少炎症介质（TNF-α、IL-1β 和 COX-2）。在缺氧的 BV-2 细胞中，T33（0.5μmol/L）减少了 TNF-α、COX-2 和 p-P38 的产生，这一作用被特异性 PPARγ 拮抗剂 GW9662（30μmol/L）预先给药所拮抗。T33（2mg/kg，ip）减轻 MCAO 诱导的炎症反应和脑梗死，这一作用被预先给予 GW9662（4mg/kg，ip）所拮抗。
T33 通过激活 PPARγ 在缺血核心和半影区发挥抗炎作用，从而发挥其神经保护作用。
