摘要
目的 利用密闭囊袋冲洗系统和人后发性白内障（PCO）囊袋模型,研究三氧化二砷（As2O3）在短时间内对PCO的防治作用.方法 实验研究.严格将时间控制在2 min时,在细胞培养条件下,四甲基偶氮唑盐比色法观察As2O3对人晶状体上皮细胞（LEC）系FHL124细胞的抑制作用;乳酸脱氢酶释放实验观察As2O3对FHL124细胞的损伤作用;荧光显微镜动态检测细胞内Ca2＋浓度的变化.建立人PCO囊袋模型,应用密闭囊袋冲洗系统观察As2O3对原代人LEC的作用.浓度反应曲线用Hill公式求IC50值.所有实验数据结果用均数±标准差表示,采用SPSS 13.0软件处理,组间差异用单因素方差分析,组间两两比较用Dunnett检验.结果 As2O3在作用仅2 min的情况下,就可以抑制人LEC和清除残留于囊袋内的人原代LEC.As2O3（10、30、100、300、1000 μmol/L）对FHL124细胞的作用呈剂量依赖性,100 μmol/L以上浓度的As2O3对FHL124细胞具有明显的毒性作用.与对照组比较,差异有统计学意义（t=5.217,P＜0.01）,半效抑制量（IC50）=130 μmol/L.乳酸脱氢酶检测显示,As2O3可影响细胞膜结构的完整性.细胞内动态Ca2＋检测显示,As2O3对细胞内Ca2＋作用呈剂量依赖性.高浓度As203可引起细胞内Ca2＋库释放,同时抑制细胞的Ca2＋内流,最终导致细胞内Ca2＋稳态的破坏而引起细胞的死亡.1×104 μmol/L As2O3作用2 min后,使Ca2＋内流的幅值及速度分别降低了（43.24±2.98）%（t=3.134,P＜0.01）和（46.27±6.01）%（t=3.521,P＜0.01）.人PCO囊袋模型显示,联合应用As2O3和密闭囊袋冲洗系统可以在2 min内清除残留于囊袋内的原代人LEC.结论 As2O3可以在短时间内彻底清除白内障手术中存留于囊袋内的人LEC,与密闭囊袋冲洗系统结合可能抑制PCO的发生.
Despite recent improvements in IOL design, posterior capsule opacification （PCO）remains a significant clinical problem after cataract surgery. The Perfect Capsule device （ Milvella, Ltd. ,Epping, Australia） permits the introduction and subsequent removal of potentially toxic agents into the closed capsular bag. The present purpose was to detect the effectiveness of arsenic trioxide （ As2O3 ） to cultured human lens cells and cells within the human capsular bag. Methods All experiments were carried out with 2 minutes exposure to As2O3. Effect of As2O3 on FHL124 cells growth was tested by MTT. Changes in cell calcium levels induced by high concentration of As2O3 were measured by real-time fluorometric single-cell digital imaging techniques after Fura-2 incorporation. In vitro human capsular bags were also tested after a sham surgery was performed using the Perfect Capsule device to form a closed system. As2O3 （10、30、100、300、1000 μmol/L ） were used with donor match-pairs treated with medium alone. （n =3 in all cases）. Ongoing observations were by phase-contrast microscopy. Cellular architecture was examined by fluorescence cytochemistry. Results As2O3 of 100 μmol/L and above inhibited the growth of FHL 124 cells in a dosedependent manner with 2 minutes exposure（ t = 5. 217, P 〈 0. 01, IC5o is 130 μmol/L）As2O3 depleted the calcium store and consequently lead to a loss of calcium signaling. Moreover, 1 × 10^4 p,mol/L As203 had a moderate effect on the calcium influx pathway,inhibition on the amplitude and rapidity is （43.24 ± 2, 98 ）% （t = 3. 134,P 〈 0. 01 ） and （46. 27 ± 6. 01 ） % （ t = 3. 521, P 〈 0. 01 ）, respectively. No cell left in the vitro human capsular bags after being treated with 1 x 104 pJnol/L As203. Conclusions As2O3 can eliminate the human lens epithelium cells remained in the capsular bag in cataract surgery, which may help to prevent PCO. The application of As203 to cells within the capsular bag for a 2 minute window using the Perfect Capsule system predicts this compound will have therapeutic benefit to humans in vivo.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2010年第11期1021-1025,共5页
Chinese Journal of Ophthalmology
基金
哈尔滨市青年科学研究基金（2005AFQXJ070）
2008年度省留学回国人员科技项目择优资助基金及黑龙江省中医药、中西医结合重点科研项目、课题、专题基金资助
关键词
白内障
砷剂
氧化物
晶体囊
上皮细胞
Cataract
Arsenicals
Oxides
Lens capsule,crystalline
Epithelial cells
