紫杉醇对视网膜色素上皮细胞功能的破坏及其潜在机制  

Damage of retinal pigment epithelial cells function by paclitaxel and its potential mechanism

作　　者：叶亚婷 窦国睿 常天芳 孙宇[1] 牛亚丽 周子义 储昭节 Ya-Ting Ye;Guo-Rui Dou;Tian-Fang Chang;Yu Sun;Ya-Li Niu;Zi-Yi Zhou;Zhao-Jie Chu(College of Life Sciences,Northwestern University,Xi an 710000,Shaanxi Province,China;First Affiliated Hospital of Northwest University,Xi an First Hospital,Xi an 710002,Shaanxi Province,China;Department of Ophthalmology,the First Affiliated Hospital of the Air Force Medical University,Xi an 710032,Shaanxi Province,China)

机构地区：[1]西北大学生命科学学院,中国陕西省西安市710000 [2]西北大学附属第一医院西安市第一医院,中国陕西省西安市710002 [3]空军军医大学第一附属医院眼科,中国陕西省西安市710032

出　　处：《国际眼科杂志》2022年第2期194-199,共6页International Eye Science

基　　金：国家自然科学基金资助项目(No.81670863,81970814);陕西省重点研发计划项目(No.2021SF-335);西安市卫健委科技计划项目(No.2020yb07);白求恩·朗沐中青年眼科科研基金(No.BJ-LM2017002J)。

摘　　要：目的:探究抗肿瘤药物紫杉醇(PTX)对人视网膜色素上皮细胞(ARPE-19)增殖、凋亡、周期、形态和视网膜外屏障的潜在毒性作用。方法:体外培养ARPE-19细胞,将细胞分为对照组(正常培养基培养)、加药物组(PTX)。不同浓度PTX(0.005、0.05、0.5、5mg/L)处理ARPE-19细胞12、24、36、48、72h,CCK8法检测不同浓度PTX不同时间点对ARPE-19细胞增殖的影响;流式细胞术检测不同浓度PTX不同时间点对ARPE-19细胞凋亡的影响及周期阻滞作用;免疫荧光观察细胞形态变化;Western blot检测凋亡相关蛋白表达及屏障功能相关蛋白表达;通过测量细胞跨上皮电阻检测药物对细胞屏障的影响。结果:PTX使ARPE-19细胞的增殖能力降低,0.005mg/L PTX处理36h,细胞增殖开始受影响;同时,PTX加速细胞凋亡且与时间、药物浓度呈依赖性,流式检测细胞周期可明显得出细胞被阻滞于G2-M期。0.05mg/L PTX处理24h后,细胞形态发生明显改变,正常细胞呈梭形或不规则形,加药物组中,细胞数相对减少且形态趋于圆形;0.05mg/L PTX破坏视网膜屏障功能,药物处理后细胞跨上皮电阻显著降低且紧密连接蛋白ZO-1、Occludin的表达较对照组显著降低(P<0.05)。Cleaved-caspase-3、Bax的表达量较对照组显著增加,Bcl-2的表达量较对照显著降低(P<0.05)。结论:PTX对ARPE-19细胞的增殖、凋亡,且依赖与时间及浓度;此外,PTX对ARPE-19细胞周期及形态均产生影响。同时PTX可破坏视网膜的屏障功能,提示抗肿瘤药物存在对视网膜潜在的毒性作用。AIM:To investigate the potential toxic effects of paclitaxel(PTX)on the proliferation,apoptosis,cell cycle,morphology,and blood-retinal barrier(BRB)of human retinal pigment epithelial cells(ARPE-19).METHODS:ARPE-19 cells were cultured in vitro and divided into two groups:Control group(Control)and drug plus group(PTX).ARPE-19 cells were treated with different concentrations of PTX(0.005,0.05,0.5,5mg/L)for a certain period of time(12,24,36,48,72h),and CCK8 assay and flow cytometry were used to detect the effects of drug on proliferation and apoptosis of ARPE-19 cells at different concentrations and time points.The same time,the cell cycle was detected by flow cytometry.Morphological changes of cells were observed by immunofluorescence.Expressions of apoptosis-related proteins and barrier function-related proteins were detected by Western blot.The effect of the drug on the cell barrier was measured by measuring the transepithelial resistance of the cells.RESULTS:PTX reduced the proliferation ability of ARPE-19 cells.After 36h of treatment with low concentration of 0.005mg/L paclitaxel,cell proliferation began to be affected.At the same time,PTX accelerated cell apoptosis was dependent on drug concentration and time.Flow cytometry showed that the cells were arrested in the G2-M phase.In addition,PTX causes significant morphological changes in cells,with normal cells fusiform or irregular.In the PTX group,the number of cells decreased and the cell shape tended to be round.PTX affected retinal barrier function,and the transepithelial resistance of cells was significantly decreased after treatment,and the expression of tight junction proteins ZO-1 and Occludin were significantly decreased compared with the control group(P<0.05).The expression levels of Cleaved-caspase-3 and Bax were significantly increased compared with the control group,while the expression levels of Bcl-2 were significantly decreased(P<0.05)and was dependent on drug concentration and time.CONCLUSION:PTX can affect the proliferation and apoptosis of AR

关 键 词：紫杉醇 人视网膜色素上皮细胞 增殖 凋亡 周期 紧密连接 

分 类 号：R73[医药卫生—肿瘤]