LPS诱导小胶质细胞活化模型的建立及评价

Establishment and evaluation of LPS stimulation on microglia activation

目的探讨不同浓度脂多糖(Lipopolysaccharides,LPS)对小胶质细胞活化作用,建立小胶质细胞活化的模型。方法不同浓度LPS(0.01,0.1,1,10,100mg·L-1)刺激原代纯化的小胶质细胞及BV2细胞株,采用MTT法检测细胞活力;采用细胞免疫化学法观察细胞形态变化。结果 LPS(1mg·L-1)可以明显激活原代小胶质细胞,但不影响细胞活力,OX-42染色显示小胶质细胞胞体变大,轴突变短变多,形似"阿米巴状";LPS(10mg·L-1)可以激活BV2细胞,且不降低细胞活力,镜下观察发现BV2细胞胞体明显增大。结论采用LPS(1mg·L-1)刺激原代小胶质细胞,LPS(10mg·L-1)刺激BV2细胞株,可以建立稳定的小胶质细胞活化模型。

Objective To explore the effect of different concentrations of LPS on microglia activation,and to establish the model of microglia activation.Methods Primary cultured microglia and BV2 microglia cells were stimulated with different concentrations of LPS（0.01,0.1,1,10,100 mg·L-1）.The cytotoxicity of LPS was measured by MTT assay,morphological changes of microglia were observed by immunocytochemical method.Results LPS（1 mg·L-1）could significantly activated primary cultured microglia but did not affect cell viability.According to the OX-42 staining pictures,after activated by LPS,the cell bodies relatively increased with multiple slender prominences like amebocytes.LPS（10 mg·L-1）could activate the BV2 microglia,but did not affect cell viability.Conclusion LPS（1 mg·L-1）stimulation of primary cultured microglia and LPS（10 mg·L-1）stimulation of the BV2 microglia can create a stable model of microglia activation.