羟基喜树碱对人胚肺成纤维细胞HFL1生长的影响

Effects of hydroxycamptothecin on human fetal lung fibroblasts

目的观察羟基喜树碱(HCPT)对体外培养的人胚肺成纤维细胞(HFL1)的增殖抑制和促凋亡作用。方法采用四甲基偶氮唑蓝法(MTT)检测不同浓度的HCPT对HFL1细胞作用24h、48h、72h的增殖抑制效果,计算细胞生长抑制率和半数抑制量(IC50),确定HCPT抑制HFL1细胞增殖的最佳给药浓度和最佳作用时间;姬姆萨染色观察细胞形态学变化;流式细胞仪分析细胞周期;琼脂糖凝胶电泳观察凋亡细胞的DNA梯度。对所得数据用SPSS 15.0软件进行统计分析。结果 HCPT对HFL1细胞增殖有明显的抑制作用,在0.008-32mg/L范围内呈时间和剂量依赖性;与对照组比较,各实验组24h、48h、72h A值及细胞生长抑制率差异有统计学意义(P<0.05,P<0.01)。HCPT作用HFL1细胞48h时的IC50为0.466mg/L,故确定48h、0.5mg/L为最佳作用时间和最佳给药浓度;0.5mg/L HCPT作用HFL1细胞24h后,G0/G1期细胞所占比例为(50.46±3.91)%;作用72h后,G0/G1期细胞所占比例为(88.19±2.13)%,与对照组比较,其余各组各期细胞所占比例差异均有统计学意义(P<0.05,P<0.01);姬姆萨染色显示,HFL1细胞形态发生明显变化;琼脂糖凝胶电泳可见明显的DNA梯度带形成。结论 HCPT对体外培养的HFL1细胞具有抑制增殖和诱导凋亡作用,可能是一种潜在的抗肺纤维化的药物。

Objective To invesligate the effect of hydroxyeamptotheein （HCVI＇） on prolifration and apoptosis of human fetal lung fibroblasts cells（ HFIA ）. Methods MTT assay using different concentrations of HCPT（0. 008,0. 016,0.031,0.063,0. 125,0.25,0.5,1,2,4,8,16.32 mg/l,）on HFLI eft＇eels for 24, 48, 72 hours and calculate the growth inhibilion rate and 50% inhibitory cnneenlration （ IC50）. The cell morpholngical changes were observed with Giemsa slaining. The cell cycle was analyzed by flnw （2ytometry. SPSS 15.0 soflware was available for the data eorivlation analysis. Results The inhibilion of HCPT on I IFIA was shown in both dose and lime dependent manner. According to the linear regression analysis, the role of time of 48h was the best and the ICs0 of HCI＇T on I Il：lA cell al the lime of 48h was 0. 466rag/L, and 48h and 0.5mg/L were used as the optimum time and concentration, The cell ratios of G0/G1 phase increased to （51）. 46±3.91 ） % for 24h and （ 88. 19 ± 2. 13 ） % for 72h after 0.5 mg/L HCVF treatment. The visible morphological changes of HFLI cells were confirmed with Giemsa staining. An olignnucleosnmal DNA ladder of era＇cumin-treated cells was demonstrated by agarose gel eleetrophoresis. Conclusion In vitro HCH＇ can significantly inhibi! prnliferation and induce apnptosis in HFLI , whieh may be helpful fnr pulmonary fibrosis.