Objective To observe the effect of lipopolysaccharide (LPS) on expression of tumor necrosis factor-ot(TNF-a) 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-a protein expression of BV-2 cells were assayed by immunohistochemical staining, and the concentrations of TNF-a 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-a protein expression (brown-yellow granules) were mainly in the BV-2 cytoplasm. The OD value of TNF-a 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, P < 0. 01). The TNF-a 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, P<0. 01). Conclusion UPS can induce the expression of TNF-a of BV-2 cells, and can change the morphology of BV-2 cells.%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24h;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.
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