高三尖杉酯碱对人视网膜色素上皮细胞增生影响的实验研究  

Experimental research on inhibition of proliferation in cultured human retinal pigment epithelial cells by homoharringtonine

作　　者：陈雯[1] 胡义珍[1] 姜发纲[1] 曾水清[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院眼科,湖北武汉430022

出　　处：《眼视光学杂志》2007年第5期317-320,共4页Chinese Journal of Optometry & Ophthalmology

基　　金：国家自然科学基金资助项目(30772382)

摘　　要：目的探讨高三尖杉酯碱(homoharringtonine,HHT)对人视网膜色素上皮细胞(retinal pigment epithelium,RPE)增生的抑制及其细胞周期的影响。方法采用MTT法检测50%细胞生长抑制率时HHT的药物浓度,并用此浓度作用于培养的人视网膜色素上皮细胞,用免疫细胞化学染色分析观察RPE细胞的Ki-67表达,流式细胞仪检测细胞周期变化。结果HHT抑制培养的人色素上皮细胞的增生与药物剂量成正比,50%细胞生长抑制率时HHT的药物浓度约为4.77mg/L,Ki-67阳性细胞率在对照组和HHT组分别为89.1%和33.8%(P<0.01),两组中阳性细胞核积分光密度值分别为60.7±5.7和29.6±7.8(P<0.01)。流式细胞仪检测细胞周期变化示HHT组G1期细胞比例由72.14%增至83.95%,G2-M期细胞比例由15.46%下降至3.18%。结论HHT可抑制RPE细胞的增生。HHT作用后RPE细胞中G0-G1期细胞增多,G2-M期细胞明显减少,HHT对G2-M期细胞有杀伤作用。HHT有望成为防治视网膜脱离复位术后发生增生性玻璃体视网膜病变(proliferative vitreoretinopathy,PVR)的一种新药。Objective To study the effect of homoharringtonine （HHT） on inhibiting proliferation and altering cell cycle in cultured human retinal pigment epithelial （RPE） cells. Methods The effect of HHT on the proliferation of RPE was measured by the MTT method at concentrations of 0.001 mg/L,0.01 mg/L,0.1 mg/L,1 mg/L, 10 mg/L,20 mg/L,and 30 mg/L. The effect of HHT on the expression of Ki-67 in cultured human RPE cells was detected by immunohistochemistry at a 5 mg/L concentration. Flow cytometry was used to analyze the cell cycle. Results The IC50 concentration of HHT was 4.77 mg/L. The relationship between the log concentration of HHT and the growth inhibition rate of RPE cells was linear. The proportion of the cells showing positive staining to Ki-67 in the control group and the HHT-treated group was 89.1% and 33.8% （P〈0.01）, respectively, and the integral optical density value for expression of Ki-67 in the two groups was 60.7±5.7 and 29.6±7.8 （P〈0.01）, respectively. The percentage of cells in the G1 phase of the cell cycle increased from 72.14% to 83.95%. The percentage of cells in the G2 phase of the cell cycle decreased from 15.46% to 3.18%. Conclusion HHT can inhibit the proliferation of RPE cells. Under its effect, RPE cells in the G0-G1 phase increased while those in the G2-M phase significantly decreased. HHT can kill RPE cells in the G2-M phase. It has the potential to become a new drug for PVR.

关 键 词：色素上皮 视网膜 细胞 培养 增生 高三尖杉酯碱 流式细胞术 

分 类 号：R285.5[医药卫生—中药学]