CAJ | 학술논문

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.
목적 이용밀폐낭대충세계통화인후발성백내장(PCO)낭대모형,연구삼양화이신(As2O3)재단시간내대PCO적방치작용.방법 실험연구.엄격장시간공제재2 min시,재세포배양조건하,사갑기우담서염비색법관찰As2O3대인정상체상피세포(LEC)계FHL124세포적억제작용;유산탈경매석방실험관찰As2O3대FHL124세포적손상작용;형광현미경동태검측세포내Ca2+농도적변화.건립인PCO낭대모형,응용밀폐낭대충세계통관찰As2O3대원대인LEC적작용.농도반응곡선용Hill공식구IC50치.소유실험수거결과용균수±표준차표시,채용SPSS 13.0연건처리,조간차이용단인소방차분석,조간량량비교용Dunnett검험.결과 As2O3재작용부2 min적정황하,취가이억제인LEC화청제잔류우낭대내적인원대LEC.As2O3(10、30、100、300、1000 μmol/L)대FHL124세포적작용정제량의뢰성,100 μmol/L이상농도적As2O3대FHL124세포구유명현적독성작용.여대조조비교,차이유통계학의의(t=5.217,P＜0.01),반효억제량(IC50)=130 μmol/L.유산탈경매검측현시,As2O3가영향세포막결구적완정성.세포내동태Ca2+검측현시,As2O3대세포내Ca2+작용정제량의뢰성.고농도As203가인기세포내Ca2+고석방,동시억제세포적Ca2+내류,최종도치세포내Ca2+은태적파배이인기세포적사망.1×104 μmol/L As2O3작용2 min후,사Ca2+내류적폭치급속도분별강저료(43.24±2.98)%(t=3.134,P＜0.01)화(46.27±6.01)%(t=3.521,P＜0.01).인PCO낭대모형현시,연합응용As2O3화밀폐낭대충세계통가이재2 min내청제잔류우낭대내적원대인LEC.결론 As2O3가이재단시간내철저청제백내장수술중존류우낭대내적인LEC,여밀폐낭대충세계통결합가능억제PCO적발생.
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