藁本内酯对脂多糖诱导神经炎症的抑制作用

Inhibitory effect of Z-Ligustilide on lipopolysaccharide-induced neuroinflammtion

目的研究藁本内酯(Lig)对细菌脂多糖(LPS)诱导的神经炎症的抑制作用。方法用1、2.5、5、10、20μmol·L^(-1)Lig作用于小鼠RAW264.7巨噬细胞,加1μg·m L^(-1)LPS刺激构建细胞炎症反应模型,采用MTT法检测细胞毒性,一步法检测NO的释放量,ELISA法检测TNF-α、IL-6的释放量,免疫细胞化学法检测i NOS的水平;将LPS注入小鼠侧脑室构建体内神经炎症模型,术前30 min及术后6 h于腹腔注射30 mg·kg^(-1)Lig,造模后24 h取大脑,用免疫组织化学法检测GFAP、TNF-α的蛋白表达。结果经Lig处理后,细胞存活率无明显变化,细胞分泌NO显著减少(P<0.05或P<0.01),且呈剂量依赖性,TNF-α、IL-6、i NOS的表达均明显下调(P<0.05);Lig能显著降低小鼠海马区GFAP、TNF-α的表达(P<0.01)。结论Lig能够抑制LPS诱导的小鼠体内外的炎症反应,提示Lig对神经炎症相关的多种疾病可能具有潜在的治疗作用。

OBJECTIVE To investigate the inhibitory effect of Z - Ligustilide （Lig） on lipopolysaccharide （LPS） - induced neuroin- flammation. METHODS The RAW264.7 macrophage cells were treated with 1,2.5,5,10,20 μmol·L^-1 Lig, then stimulated with 1 μg·mL^- 1 LPS to construct inflammatory cell model. The cell viability was determined through MTT assay ; the nitric oxide （ NO ） release was detected by one - step method ; the expression of tumor necrosis factor - α （ TNF - α） ; interleukin 6 （ IL - 6 ） were detected with ELISA;and inducible nitric oxide synthase （iNOS） expression was tested by immunocytoehemistry. The neuroinflammatory model was set up by injecting LPS directly into the right lateral ventricle of mice. 30 mg·kg^-1 Lig was intraperitoneally administered, 30 min before and 6 h following LPS injection. Twenty - four hours after LPS administration, the brains were removed, and immunohistochemistry was used to detect the expression of glial fibrillary acidic protein （GFAP） and TNF - α. RESULTS After Lig treatment, the cell viability had no significant effect,the secretion of NO was dose - dependent inhibited （ P 〈 0.05 or P 〈 0.01 ）, and the expression of TNF -α, IL - 6 and iNOS was clown regulated （ P 〈 0.05 ）. Lig decreased effectively immunoreactivity of GFAP and TNF - α in the hippocampus （P 〈 0.01 ）. CONCLUSION Lig exerts a potent inhibitory action on LPS - induce inflammatory responses in in vitro and in vivo mice, and has therapeutic potential in neuroinflammation -associated disorders.