表没食子儿茶素没食子酸酯减轻LPS诱导新生大鼠原代神经胶质细胞损伤

Epigallocatechin-3-gallate( EGCG)reduces the LPS-induced injury of newborn rat primary cultured glial cells

目的观察表没食子儿茶素没食子酸酯(EGCG)对细菌脂多糖(LPS)所致大鼠神经胶质细胞炎症的保护作用。方法取新生乳鼠原代神经胶质细胞培养,用LPS激活小胶质细胞引起炎性反应。高效液相色谱检测细胞兴奋性/抑制氨基酸含量,用酶联免疫吸附实验(ELISA)和蛋白免疫印记试验(Western blot)检测炎性因子蛋白含量。结果 LPS激活神经小胶质细胞,大幅上调TNF-α、IL-1β及IL-8炎性因子和升高i NOS蛋白质水平(P<0.05),EGCG明显抑制这些炎性因子的过度产生(P<0.05),同时还显著降低Glu和升高γ-GABA的浓度(P<0.05)。结论EGCG能减弱LPS引起的体外培养神经胶质细胞的炎性反应。

Objective To determine the effects of EGCG on lipopolysaccharide（ LPS）-induced neuroinflammation and investigate the role of neuroprotection mediated by EGCG. Methods Primary cultures of rat gliacyte were used as an in vitro model to examine the effects of EGCG on LPS-induced neuronal damage. The intracellular Glu and γ-GABA were quantified via HPLC. Then the protein level of TNF-α,IL-1 β and IL-8 was determined by ELISA and Western blot assay. Results Compared with the control group,LPS apparently induced the production of intracellular ROS（ P 〈0. 05） and released the TNF-α,IL-1 β and IL-8 in the primary cultures supernatant（ P 〈0. 05）. Compared with the LPS group,EGCG significantly attenuated the release of TNF-α,IL-1 βand IL-8（ P 〈0. 05） and the level of i NOS protein（ P〈 0. 05）. LPS apparently induced the production of intracellular ROS（ P〈 0. 05） and released the TNF-α,IL-1 β and IL-8 in the primary cultures supernatant（ P 〈0. 05）. EGCG significantly attenuated the release of TNF-α,IL-1β and IL-8（ P 〈0. 05） and the level of i NOS protein（ P 〈0. 05）,and rugulated the concentration of Glu/γ-GABA（ P 〈0. 05）. Conclusions EGCG is effectivein protecting hosts against LPS-induced neuroinflammation through anti-inflammatory effects and regulating extracellular Amino acid levels.