摘要
目的:观察TNP-470对人脐静脉内皮细胞系ECV-304细胞生长的影响.方法:采用MTT、细胞生长曲线、倒置显微镜形态学观察、荧光染色、流式细胞术等方法观察不同浓度TNP-470对ECV-304细胞形态、细胞增殖和细胞周期的影响.结果:MTT结果显示:在低浓度(0.2μg/L)时,TNP-470对ECV-304增殖无明显抑制作用,从2μg/L开始产生抑制作用,且该抑制作用具有量效关系,IC50值为305μg/L;生长曲线:20,200,2000μg/LTNP-470处理细胞后,细胞增殖被抑制,且具有剂量—时相依赖性;倒置相差显微镜下观察:各时间段对照组细胞生长状态正常,排列整齐呈铺路石样;从2μg/L开始出现细胞增殖抑制现象,随作用时间的延长和浓度增高抑制作用更为明显;2000μg/L组见少部分脱落、悬浮细胞;流式细胞仪分析经20,200,2000μg/L的TNP-470作用24,36h后,细胞被阻滞在G0/G1期,G2/M和S期的细胞数减少(P<0.05),细胞分裂受抑制,增殖指数降低。结论:TNP-470能使ECV-304细胞周期阻滞在G0/G1期,对细胞增殖具有抑制作用。
AIM： To analyze the inhibitory effect of TNP-470 on the growth of human umbilical vein cells （ECV-304 cells） in vitro.
METHODS： ECV-304 cells were incubated with different concentration of TNP-470 （0.2, 2, 20, 200, 2 000μg/L）. The cell morphology, proliferation and cell cycle were observed by MIF assay, inversion microscope, fluorescence staining and flow cytometry.
RESULTS： MIF assay showed the inhibition occurred at 2μg/L or higher concentration in a dose-dependent manner. The IC50 value was 305μg/L. Cells proliferation were significantly inhibited （P〈0.01） at the concentration of 20, 200, 2 000μg/L of TNP-470 in a dose-phase-dependent manner. Observation under the microscope showed the cells grew well at all the stages and lined up like slabstones. Cells growth inhibition occurred at 2μg/L and maximum inhibition occurred at 2 000μg/L group. After incubation with 20, 200, 2 000μg/L TNP-470 for 24h. 36h, cells cycle were blocked at G0/G1 stage by the detection of flow cytometry. The cell numbers in G2、S stage were significantly decreased, with lower proliferation index.
出处
《国际眼科杂志》
CAS
2005年第6期1146-1149,共4页
International Eye Science
