丁基苯酞对脂多糖活化的小胶质细胞炎症介质释放的影响

Effects of d1-3-n-butylphthalide on the release of inflammatory mediators in lipopolysaccharide-activated microglia cells

目的·观察丁基苯酞(NBP)对脂多糖(LPS)激活后的小胶质细胞炎症介质释放的影响。方法·原代培养大鼠脑皮质小胶质细胞和神经元,将接种后的小胶质细胞分为5组,即空白对照组、LPS激活组、溶剂+LPS组、0.01 mmol/L NBP+LPS组以及0.1 mmol/L NBP+LPS组。在LPS作用前2 h用NBP对小胶质细胞进行预处理,经LPS刺激24 h后在倒置显微镜下观察小胶质细胞形态;同时用酶联免疫吸附测定(ELISA)法检测各组小胶质细胞释放的炎症因子肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)及白介素-10(IL-10)含量;用硝酸还原法检测一氧化碳(NO)含量;经四甲基偶氮唑盐(MTT)法检测经各组作用后的神经元存活率。结果·NBP预处理后的小胶质细胞经LPS刺激后仍呈激活形态,NBP不影响激活的小胶质细胞释放炎症因子TNF-α、IL-1β以及IL-10,但可抑制其释放炎症介质NO,提高神经元存活率,并呈现剂量依赖性。结论·NBP可减轻小胶质细胞经LPS活化后的神经炎症反应,并具有剂量依赖性的神经保护作用。

Objective · To investigate the effects of d1-3-n-butylphthalide (NBP) on inflammatory mediators released by microglia cells after beingactivated with lipopolysaccharide (LPS). Methods · Microglia cells and neurons were obtained from primary cultured rat cortices. Microglia cells weredivided into five groups, including the blank control group, the LPS activated group, the solvent plus LPS group, the 0.01 mmol/L NBP plus LPS group,and the 0.1 mmol/L NBP plus LPS group. Microglia cells were pretreated with NBP 2 h before LPS stimulation. The morphology of microglia cells wasobserved under inverted microscope after being stimulated with LPS for 24 h. The levels of inflammatory factors, including tumor necrosis factor α (TNF-α),interleukin 1β (IL-1β), and interleukin 10 (IL-10), were measured with enzyme linked immunosorbent assay (ELISA) method and the content of carbonmonoxide (NO) was measured using nitrate reductase method. The survival rate of neurons was detected by MTT method. Results · NBP pretreatedmicroglia cells remained the activated form after LPS stimulation. NBP did not affect the release of TNF-α, IL-1β, and IL-10 by activated microglia cells,but inhibited the production of inflammatory mediator NO, thus improved the survival rate of neurons in a dose-dependent manner. Conclusion · NBP canattenuate the LPS-induced neuroinflammatory responses in microglia cells and show the dose-dependent neuroprotection.