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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 observe the inhibitory effect of Tet (Tet) on the growth of cultured rabbit conjunctival fibroblasts (RCFs) in vitro and to explore the possible mechanism of this effect and to provide an effective method for the prevention and treatment of various eye diseases associated with fibroblast proliferation Drug. Methods: MTT assay was used to determine the inhibitory effect of different concentrations of Tet on RCF. The cytotoxicity of different concentrations of Tet to RCF was observed by cytometry. The effect of Tet on RCF cell cycle was detected by flow cytometry. RESULTS: Tetraphenol at concentrations of 0 16, 0 8, 4, 20 0 and 10 0 μg / ml significantly inhibited the growth of RCF after 48 hours of administration. Under the above effects of different concentrations of Tet, The cells were counted on day 1, 3, 5 and 7, and it was found that the inhibition of Tet on RCF was time-and dose-dependent. Flow cytometry showed that RCF treated with 2 μg / ml Tet for 48 h showed an increase of 13 3% in G1 phase, 95 5% in S phase, and 42.3% in G2 phase. Conclusion: Tetrandrine can inhibit the growth of RCF with time-dose effect. Tet affects the population of RCF cell cycle phases and inhibits the transition from G1 phase to S phase, which in turn prevents the cell from transitioning to G2 phase and thus inhibits cell proliferation.