蝮蛇抗栓酶对人翼状胬肉成纤维细胞增殖效果的影响

The effects of ahylysantinfarctase on the proliferation of human pterygium fibroblasts

目的 :研究国产蝮蛇抗栓酶 (ahylysantinfarctase ,AF)对体外培养的人翼状胬肉成纤维细胞 (humanpterygiumfibroblasts ,HPF)增殖效果的影响。方法 :采用切除的翼状胬肉组织进行体外成纤维细胞培养 ,将传代的第 4～第 6代人翼状胬肉成纤维细胞与不同浓度的蝮蛇抗栓酶 ( 0 .1U/ml～0 .0 0 2 56U/ml)分别处理 4 8h ,并设对照组 ,通过噻唑兰比色法 (MTT法 ) ,在酶联免疫测定仪上测 570nm处吸光值 ,计算出在不同浓度AF条件下的净增抑制率。结果 :AF在 0 .0 4U/ml以上使细胞增殖率显著下降 ;当AF浓度为 0 .1U/ml时 ,其细胞增殖抑制率达 91 .74 %左右 ,镜下很少见到正常活性细胞。AF在 0 .0 0 2 56U/ml以上使部分细胞生长受影响。其 50 %抑制率浓度 (ID5 0 )为 0 .0 1 6U/ml。结论 :AF对HPF增生有抑制作用 ,并且其抑制作用呈剂量依赖型 ,但在高浓度(≥ 0 .1U/ml)时可能存在细胞毒性作用。

Objective:To investigate the effects of ahylysantinfarctase (AF) on the proliferation of human pterygium fibroblasts (HPF) cultured in vitro.Methods:Pterygium biopsies were excised from patients. The primary culture and subculture of HPF were established in vitro. Different concentrations of AF (0.1 U/ml～0.00256 U/ml) were added to the HPF in 4～6 passages over 48 hours and then compared to the control group. The effects of AF on the proliferation of HPF were determined by MTT colorimetric assay.Results:The addition of AF to the HPF caused significant inhibition of the HPF proliferation cultured in vitro,which was dose dependent. The median inhibition dose (ID50) of AF was 0.016 U/ml. But when the concentration of AF was 0.1 U/ml,the inhibition rate on the proliferation of HPF reached 91.74%,and histological observation showed few normal HPF. These findings showed that there may be cytotoxicity to HPF when the concentration of AF is higher than 0.1 U/ml.Conclusion:AF can significantly inhibit the proliferation of HPF cultured in vitro with a dose dependent function (ID 50 =0.016 U/ml). But there may be cytotoxicity to HPF when the concentration of AF is above 0.1 U/ml.