汉防己甲素和艾氟龙对兔角膜基质细胞抑制作用的对比研究

An investigation of inhibition of tetrandrine(Tet) and fluorometholone (FML) on rabbit corneal stroma cells (RCSCs)

目的:探讨比较汉防己甲素(Tet)和艾氟龙对离体兔角膜基质细胞的抑制作用及其作用机制。方法:选用原代及传代兔角膜基质细胞,Tet组加入含不同质量浓度Tet的培养液,艾氟龙组加入不同质量浓度艾氟龙的培养液,对照组加入含等量PBS的培养液。采用四甲基偶氮唑蓝(MTT)比色法检测对细胞增生的抑制;免疫细胞化学法检测E2F1的表达状况。结果:Tet和艾氟龙对角膜基质细胞生长都具有抑制作用,各时间点IC50Tet均1低于艾氟龙。免疫细胞化学结果显示,Tet组、艾氟龙组和对照组均有E2F1蛋白的表达,但Tet组和艾氟龙组中E2F1蛋白的表达均减弱,Tet组中E2F1蛋白的表达减弱更明显。结论:Tet和艾氟龙对角膜基质细胞的增生均有抑制作用,且Tet对角膜基质细胞的抑制作用强于艾氟龙。Tet和艾氟龙可能通过降低E2F1的表达使细胞周期停滞而发挥其对角膜基质细胞的抗增生作用。

Objective：In this study,we discussed the effect of Tet and FML on inhibition of RCSCs and explored the mechanism.Methods：The primary cultured and subcultured RCSCs were exposed to the culture media containing 20μl PBS and different concentrations of Tet and FML.The inhibitory characteristics of Tet and FML to the proliferation of cells were measured with methyl thiazolyl thiazolium（MTT） assay.The changes of E2F1 were measured by immunocytochemistry.Results：After the treatment of Tet and FML,RCSCs showed the marked inhibition in 24,48 and 72 hours（P0.01） in a dose-depend manner and the IC50 of Tet at each time point was lower than that of FML.The brown positive expression of E2F1 in RCSCs was displayed in caryon.In Tet groups and FML groups,the expression of E2F1 decreased.The staining of the positive cells in Tet groups was weaker than that in FML groups.Conclusion：Tet and FML have inhibitory effect on the proliferation of RCSCs.The inhibitory effect of Tet is more dominant than that of FML.Tet and FML may exert the antiproliferative effect on RCSCs by ceasting cell cycle with E2F1.