汉防己甲素对体外培养的兔结膜成纤维细胞抑制作用的实验研究

Inhibitory effect of tetrandrine on the cultured rabbit conjunctival fibroblasts

目的 :观察汉防己甲素 (Tet)对体外培养的兔结膜成纤维细胞 (RCF)生长的抑制作用 ,探讨其作用的可能机制 ,为防治与成纤维细胞增殖相关的多种眼病提供一种有效的药物。方法 :应用MTT法测定不同浓度的Tet对RCF的抑制作用 ;用细胞计数法观察不同浓度的Tet在不同时间下对RCF的生长抑制作用 ;用流式细胞仪检测Tet对RCF细胞周期的影响。结果 :给药 4 8小时后 ,终浓度为 0 16、0 8、4、2 0、10 0 μg/ml的Tet对RCF的生长均有明显抑制作用 ;且在上述不同浓度的Tet作用下 ,于 1、3、5、7天计数细胞 ,发现Tet对RCF的抑制具有时间、剂量关系。流式细胞仪显示 ,RCF经 2 μg/mlTet处理 4 8h后 ,G1期细胞增加了 13 3 % ,S期细胞下降了 9 5 5 % ,G2 期细胞下降了 4 2 3 %。结论 :汉防己甲素可抑制RCF生长 ,有时间剂量效应。Tet影响RCF细胞周期各时相群体 ,抑制G1期向S期的过渡 ,进而阻止细胞向G2 期的过渡 ,从而抑制细胞增殖。

Objective:To investigate the inhibiting effects of tetrandrine (Tet) on the proliferation of the rabbit conjunctival fibroblasts (RCF) in vitro and the mechanism of inhibition.Methods:Cultured RCF were exposed to different concentrations of Tet solution and investigated by MTT assay after exposure to Tet 48 hours.Using Abbe Zeiss counting observe the effect of Tet on cells growth curve at different times and in different concentrations. The changes of cell cycle were measured by flow cytometer after treated with Tet (2μg/ml) on day 2 Results:Tet(0 16 100μg/ml) significantly suppressed the proliferation of RCF in vitro and showed a dose dependent and time dependent tendency after day1,day3,day5 and day 7 of exposure to Tet.The cells treated with Tet (2 μg/ml) were increased 13 3% in G 1 phase,decreased 9 55% in S phase and 4 23% in G 2 phase respectively.Conclusions:Tet has an effective inhibition on the proliferation of RCF and may supply a potential prospect in future clinical application.