脂多糖对培养的人角膜基质细胞生长的影响  

The effect of bacterial lipopolysacchride to human cultural corneal stromal cells

作　　者：姜宏[1] 许海洋[2] 郝继龙[1] 王爽[1] 

机构地区：[1]吉林大学中日联谊医院眼科,吉林长春130031 [2]吉林大学第一医院神经外科,130011

出　　处：《中国实验诊断学》2009年第3期348-350,共3页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金资助项目(编号:30772384)

摘　　要：目的观测细菌脂多糖对人角膜基质细胞的影响,探索其应用于人角膜基质细胞实验研究的安全浓度范围,为人角膜基质细胞的炎症相关实验研究奠定基础。方法体外培养人角膜基质细胞,取第3-5代细胞鉴定后用于实验。采用四氮唑盐代谢法(MTT),检测不同浓度和作用时间脂多糖对培养的人角膜基质细胞增殖的影响;采用相差显微镜观察及0.4%台盼兰染色法,检测脂多糖对培养的人角膜基质细胞存活率的影响与其浓度和作用时间的关系。结果脂多糖对培养的人角膜基质细胞的影响与浓度和作用时间有关。脂多糖在浓度高于一定水平时可以引起细胞增殖和存活率的下降,浓度相同时作用时间越长下降越明显。脂多糖在浓度1 000 ng/ml以下作用24 h不会对细胞增殖和存活率产生影响。结论脂多糖在一定浓度范围内不引起细胞毒性,可以用于炎症条件下培养的人角膜基质细胞功能的实验研究。Objective To observe the effect of bacterial lipopelysacchride to human cultured corneal stromal cells, to explore the safe range of density in the experimental study of human corneal stromal cells. So it can establish the foundation of the study of the inflammafion of human corneal stromal cells.Methods To culture the human corneal stromal cells in vitro, chose the 3 - 5 generation and identify for the test. By tetrazolium-based metabolic technique（MTT）, to detect the effect of different lipopolysacchride density and action- time to the proliferation of human cultured corneal stromal coils;by phase contrast microscope to observe the results and by 0.4% trypan blue staining method to study the reation between the effect of differont lipopolysaechride density and action-time and the viability of human cultured corneal stromal coils.Results The effect of lipepolysacchride to human cultured corneal stromal cells is related with the density and action-time.Beyond certain density,the lipopolysacchride can cause decrease of coll proliferation and viability. With same density,the longer action-time,the effect will be more obvious. The lipopolysacchride will not effect the proliferation and viability under the density of 1 000 ng/ml. Conclusion The iipepolysacchride will not produce cytotexicity in certain range of density. It can be used in the experimental study of human corneal stromal cells cultured in the inflammation condition.

关 键 词：角膜基质细胞 脂多糖 

分 类 号：R772.2[医药卫生—眼科]