摘要
目的探讨脂多糖对BV-2细胞的形态及肿瘤坏死因子-α（TNF-α）表达的影响。方法 DMEM高糖培养基中培养的BV-2细胞平均分为空白对照组与脂多糖处理组。脂多糖处理组BV-2细胞采用含100ng/ml脂多糖的培养基培养24 h;空白对照组用PBS替代脂多糖。采用免疫组化染色法观察BV-2细胞的形态及TNF-α蛋白的表达,采用酶联免疫吸附试验（ELISA）法检测BV-2细胞培养上清液中TNF-α的浓度。结果空白对照组BV-2细胞形态呈上皮细胞样,每个细胞有2~3个突起,突起较细长,细胞核较小。脂多糖处理组BV-2细胞多数聚集成团,大部分胞体肥大、饱满,细胞核大而圆,核仁明显,胞体突起短小,形态均具有小胶质细胞激活后形态特征。BV-2细胞中TNF-α蛋白的阳性表达均主要在胞浆中,为棕黄色颗粒。空白对照组BV-2细胞TNF-α蛋白阳性表达产物的OD值（5.46±0.87）明显低于脂多糖处理组（53.01±5.72）,差异有统计学意义（t=26.01,P〈0.01）。空白对照组BV-2细胞培养上清液中TNF-α浓度为（218.13±24.10）pg/ml,脂多糖处理组BV-2细胞培养上清液中TNF-α浓度为（1098.69±72.18）pg/ml,两组间比较差异有统计学意义（t=23.14,P〈0.01）。结论脂多糖可显著诱导BV-2细胞系TNF-α的表达,同时可显著改变BV-2细胞的形态。
Objective To observe the effect of lipopolysaccharide（LPS） on expression of tumor necrosis factor-α（TNF-α） and cell form of BV-2 cells.Methods BV-2 cells cultured in DMEM high glucose mediums were divided into control group and the LPS treated group.BV-2 cells of LPS treated group were treated with 100 ng/ml LPS in culture medium for 24 h,while the control group were treated with PBS instead of LPS.The morphology and TNF-α protein expression of BV-2 cells were assayed by immunohistochemical staining,and the concentrations of TNF-α in BV-2 cell culture supernatant were detected by enzyme-linked immunosorbent assay（ELISA）.Results The morphology of BV-2 cells in control group were skin cell-like,with each cell had 2-3 processes,and the processes were slender,the cell nuclei was smaller.The BV-2 cells of LPS treated group were mostly gathered into a group.Most of the cells with cell body hypertrophy and full,nuclei large and round,nucleoli prominent and cell processes short;which just liked the forms of activated microglia morphology.The TNF-α protein expression（brown-yellow granules） were mainly in the BV-2 cytoplasm.The OD value of TNF-α protein expression in control group（5.46 ± 0.87） was significantly lower than that in LPS treated group（53.01 ± 5.72）,and the difference was statistically significant（t=26.01,P0.01）.The TNF-α concentration in BV-2 cell supernatant in control group was（218.13 ± 24.10） pg/ml,and which in LPS treated group was（1098.69 ± 72.18） pg/ml;the difference between the two groups was statistically significant（t=23.14,P0.01）.Conclusion LPS can induce the expression of TNF-α of BV-2 cells,and can change the morphology of BV-2 cells.
出处
《临床神经病学杂志》
CAS
北大核心
2011年第5期365-367,共3页
Journal of Clinical Neurology
