脂多糖刺激RAW264．7和Ana-1形态改变及细胞因子表达的差异

Comparison between two kinds of murine macrophages-Ana-1 and RAW264. 7 cell in appearance and eytokines expression

目的比较两株小鼠来源的巨噬细胞株RAW264．7和Ana-1经脂多糖（lipopolysaccharide，LPS）诱导后在形态及细胞因子表达等方面的差异。方法MTT法检测不同终浓度（0．1mg／L、1mg／L、10mg／L、100mg／L）LPS作用后两株细胞增殖活力，确定最佳作用浓度。选择1mg／L．LPS刺激RAw264．7和Ana-1细胞，ELISA法分别于0h、4h、8h、12h、24h检测两株细胞肿瘤坏死因子α（TNF-α）、白介素1β（IL-1β）、IL-6和IL-10的表达量。结果LPS终浓度为1mg／L时两株细胞存活率分别为（162．05±28．14）％、（159．92±20．43）％，显著大于同细胞其他浓度组别（P〈0．01）；1mg／L LPS刺激后，两株细胞各细胞因子表达均为随时间呈先增多后减少趋势，峰值出现时间RAW264．7细胞早于Ana-1细胞。TNF—α的表达4h时RAW264．7细胞高于Ana-1（P〈0．01），8h、12h时低于后者（P〈0．05）；IL-18的表达各时间点比较RAW264．7均高于Ana-1（P〈0．05）；IL-6的表达各时间点比较RAW264．7均低于Ana-1（P〈0．05）；IL-10的表达于刺激后4h，RAW264．7细胞高于Ana-1细胞（P〈0．05）。结论当LPS浓度为1mg／L时，Ana-1与RAW264．7细胞存活率均最强；经LPS刺激后，RAW264．7细胞TNF-α、IL-6、IL—1β、IL-10表达高峰早于Ana-1细胞，各细胞因子表达量各有侧重。研究者进行炎症相关实验时对Ana-1和RAW264．7的选择需考虑两者之间的差异。

Objective To compare the morphologic change and cytokines expression in RAW264. 7 and Ana-1 stimulated by lipopolysaecharide（LPS）. Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS（0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L）. MTT was performed to evaluate the proliferation ability of cells. Two kinds of ceils were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6 ,IL-10 in culture medium were detected by ELISA in different times（0 h,4 h,8 h, 12 h,24 h）.Results The survival rates of RAW264.7 and Ana-1 （162. 05±28.14）% and （159.92±20.43）% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner （increased first and decreased finally）. The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.