Department of Neurology, Affiliated Dongtai Hospital of Nantong University, Dongtai, 224200, Jiangsu, People's Republic of China.
Inflamm Res. 2013 Nov;62(11):929-40. doi: 10.1007/s00011-013-0631-2. Epub 2013 Sep 26.
To investigate whether Nischarin participated in neuronal apoptosis induced by neuroinflammation and via the phosphatidylinositol 3-kinase (PI3K) and PKB-dependent pathway.
Use of male Sprague-Dawley rats, rat pheochromocytoma (PC12), and murine microglial cells (BV-2). Treatment lipopolysaccharides (LPS) were injected into the brain lateral ventricle of the rat. The BV-2 cells were treated by LPS. The PC12 cells were pretreated by or not pretreated by conditioned media and siRNA.
Western blotting was used for analyzing the expression level of Nischarin, pAKT, BAD and Bcl-2. Immunohistochemistry and immunofluorescence were used to perform the morphology and localization of Nischarin. The siRNA could down-regulate the protein level of endogenous Nischarin.
The expression level of Nischarin was elevated after LPS injection; meanwhile, Nischarin was located in the neuron. Nischarin was involved in regulating the PI3K/PKB patway.
Nischarin might be involved in mediating the process of PI3K/PKB pathway-dependent neuronal apoptosis. After the silencing of Nischarin in cultured PC12 (pheochromocytoma) by siRNA, these results showed that it would induce a reduction of pAKT and Bcl-2 proteins expression; meanwhile, it induces an increase of BAD and active caspase-3.
探讨 Nischarin 是否参与神经炎症诱导的神经元凋亡，并通过磷脂酰肌醇 3-激酶（PI3K）和 PKB 依赖性途径发挥作用。
雄性 Sprague-Dawley 大鼠、大鼠嗜铬细胞瘤（PC12）和鼠小胶质细胞（BV-2）。用脂多糖（LPS）处理大鼠侧脑室。用 LPS 处理 BV-2 细胞。用条件培养基和 siRNA 预处理或不预处理 PC12 细胞。
Western blot 用于分析 Nischarin、pAKT、BAD 和 Bcl-2 的表达水平。免疫组织化学和免疫荧光用于进行 Nischarin 的形态和定位。siRNA 可以下调内源性 Nischarin 的蛋白水平。
LPS 注射后 Nischarin 的表达水平升高；同时，Nischarin 位于神经元中。Nischarin 参与调节 PI3K/PKB 通路。
Nischarin 可能参与介导 PI3K/PKB 通路依赖性神经元凋亡过程。用 siRNA 沉默培养的 PC12（嗜铬细胞瘤）中的 Nischarin 后，结果表明它会导致 pAKT 和 Bcl-2 蛋白表达减少；同时，它会导致 BAD 和活性 caspase-3 增加。
