摘要
目的研究水蛭渗滤液对酪氨酸蛋白激酶（PTK）介导的牛视网膜色素上皮（RPE）细胞跨膜信号转导途径的影响。方法（1）MTT比色法筛选促牛RPE细胞增殖作用最强的血小板源性生长因子（PDGF）浓度;（2）16mg/ml水蛭渗滤液与0.02μg/mlPDGF以3种加药方式分别孵育RPE细胞15分钟后,采用免疫沉淀,SDS-PAGE电泳,Western-blot检测转录因子STAT3磷酸化水平。结果（1）0.02μg/mlPDGF促RPE细胞增殖作用最佳（p〈0.05）;（2）16mg/ml水蛭渗滤液在3种加药方式下均能不同程度抑制细胞内PTK活性,降低转录因子STAT3的磷酸化水平。结论水蛭渗滤液能阻断PTK介导的牛RPE细胞内STAT3下游通路的信号传递。
Objective To research the influence of percolation filtration of Hirudo on bovine retinal pigment epithelial（RPE） cells＇ signal transduction that is guided by protein tyrosine kinase（PTK）, Methods （1）The method of MTT was used to select the proper concentrations of platelet-derived growth factor（PDGF） that proliferate RPE cells greatly. （2）The choosen 16mg/ml percolation filtration of Hirudo and 0.02 ug/ml PDGF were used to incubate RPE cells for 15 minutes with three different modes .Then the bovine RPE cells were splitted. The phosphorylated level of transcription factor-STAT3 in the cells was studied through immunoprecipiration, SDS-PAGE and Western-blot. Results （1）When the concentration of PDGF solution was 0.02ug/ml, the proliferation effect was the most strong （p〈0.05）. （2） The activity of PTK in the cells was inhibited in different degrees by 16mg/ml percolation filtration of Hirudo which was added with three different modes, and the phosphorylated level of STAT3 was reduced. Conclusion Percolation filtration of Hirudo can interdict the cells＇ signal transduction after STAT3,
出处
《中国实用眼科杂志》
CSCD
北大核心
2007年第5期549-551,共3页
Chinese Journal of Practical Ophthalmology
基金
国家自然科学基金资助项目,基金编号30371796
