多西他赛对人晶状体上皮细胞的抑制作用  

Inhibitory effect of docetaxel on proliferation of human lens epithelial cells

作　　者：易少凌[1] 施斌[1] 李婉文[1] 徐礼剑[1] 赵春顺[1] 

机构地区：[1]中山大学药学院,广州510006

出　　处：《中华实验眼科杂志》2011年第1期32-37,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：广东省科技计划项目（20078060401057）

摘　　要：背景一些已知的抑制人晶状体上皮细胞（LECs）增生的药物由于严重的不良反应限制了其临床应用，寻找高效、安全的抑制LECs增生的药物是防治晶状体后囊膜混浊（PCO）的研究热点。目的探讨多西他赛对LECs增生的影响，并与盐酸表柔比星、盐酸吡柔比星和雷替曲塞的作用进行比较。方法对永生系人LECs细胞株SRAOI／04进行培养及传代，将不同浓度的多西他赛、盐酸表柔比星、盐酸吡柔比星和雷替曲塞加入LECs中分别作用24、48、72h，以MTT法评价不同浓度的多两他赛对人LECs增牛的抑制作用并与其他药物进行比较。用流式细胞术分析不同浓度的多西他赛作用48h后对LECs细胞周期的影响，采片jAnnexinV-FITC／PI标记法和蛋白印迹分析法评估不同浓度的多西他赛作用48h后LECs细胞bcl-2蛋白的表达和凋亡情况。结果8～519pmol／L多西他赛组LECs的增生率为100％，LECs形态正常，而66nmol／L多西他赛组干预48h和72h后LECs的增生率分别为34．7％和27．4％，LECs形态出现异常。23．22～523．56μmol／L雷替曲塞对人LECs的增生无明显抑制作用。多西他赛作用48h和72h时，半数抑制量（IC50）明显低于盐酸吡柔比星和盐酸表柔比星。多西他赛作用LECs48h后，随着多西他赛浓度的增高，处于G2／M期的LECs百分数明显增加，各组的总体差异有统计学意义（F=2633．05，P〈0．01）；8．3nmol／L和266nmol／L多西他赛浓度组干预48h后，LECs的凋亡率分别为22．4％和27．9％，较细胞对照组升高，差异均有统计学意义（χ2=20．00，P〈0．01；χ2=42．68，P〈0．01）。Westernblot检测表明不同浓度多西他赛干预后bcl-2蛋白在LECs中的表达条带均较对照组淡，8．3nmol／L多西他赛组bcl-2蛋白表达降低更为明显。结论多西他赛、盐酸表柔比星和盐酸吡柔比星均可抑制人LECs的增生，而雷替曲塞对人LECs的增生无明显的抑�Background Some drugs with inhibitory effect on the proliferation of lens epithelial cells have a limiting application in clinic because of their adverse response. To screen the effective and less side-effect drug for supressing LECs growth is very inportant for the prevention and treatment of after cataract. Objective This study was to explore the effects of docetaxel on LECs growth and compare its role with epirubicin hydroch/oride,pirarubicin hydrochloride and rahitrexed. Methods Immortalized huraan LECs line （SRA01/04） were cultured and passaged. Different concentrations of docetaxel, epirubicin hydrochloride, pirarubiein hydrochloride and rahitrexed were added into the medium respectively for 24,48 and 72 hours. The proliferation of LECs was detect by MTT. Flow eytometry analysis was used to analyze the influence of different concentrations of docetaxel on cellular cycle at 48 hours after addition of doeetaxel,and Annexin V-FITC/PI marking method was used to assesse the apoptosis of LECs under the action of docetaxel. Expression of bcl-2 protein in LECs was evaluated by Western blot. Result The growth rate of LECs was 100% in 8-519 pmol/L docetaxel groups with the normal cell shape. Majority of abnormal cells and low growth rate were found in 66 nmol/L docetaxel group at 48 and 72 hours. The IC50 of docetaxel was lowest in 48 and 72 hours in docetaxel group in comparison to epirubicin hydrochloride and pirarubicin hydrochloride. However,no evident inhibition on LECs growth in 23.22-523.56 μmol/L of raltitrexed. At 48 hours,the percentage of LECs in G2/M phase increased as the ascente of concentration of docetaxel,showing a significant difference among 4 groups （F= 2633.05 ,P〈O. 01 ）. The percentage of early apoptotic cells increased to 22.4% （Xz = 20.00,P〈O. O1 ） and 27.9% （X2 = 42.68,P〈0.01 ） from normal control 3. 1% at 48 hours after LECs exposed to 8.3 nmol/L and 266 nmol/L doeetaxe. The expression of bcl-2 protein in LECs was obviously weakened after addition of docetaxel, especia

关 键 词：晶状体上皮细胞 后囊膜混浊 多西他赛 盐酸表柔比星 盐酸毗柔比星 雷替曲塞 

分 类 号：R776.1[医药卫生—眼科]