PEDF对人晶状体上皮细胞生长调控的体内外研究.docVIP

PEDF对人晶状体上皮细胞生长调控的体内外研究 刘恬1，项道满1，江福能2 作者单位：1、广州市妇女儿童医疗中心眼科 广州 510广州医科大学附属广州市第一人民医院广东省临床分子医学及分子诊断重点实验室 广州 510180 fifaspring@ 基金项目：广州市卫生局项目资助（201102A213035） 摘要：目的 探讨在体内及体外色素上皮衍生因子（PEDF）对人晶状体上皮细胞（LEC）生长的调节作用。 方法 体内研究，用ELISA法检测年龄相关性白内障患眼房水PEDF浓度，术中取前囊膜标本检测LEC密度，分析PEDF水平与LEC数量、白内障类型的关系；体外研究，加PEDF培养人晶状体上皮细胞株（HLE-B3），CCK8法检测细胞生长活性，流式细胞仪通过AnnexinV-FITC/7-AAD双染法检测细胞周期和凋亡变化。 结果 120例白内障患眼房水PEDF浓度为156.2±41.2ng/ml，中央区前囊下LEC密度随房水PEDF水平下降而降低（r=0.556，P0.01），控制年龄因素后仍呈正相关（r=0.387，P0.05），皮质型白内障房水PEDF浓度较核硬化型更低（P0.05）；体外培养的LEC细胞株加入PEDF后，细胞生长明显加快（F=187.33，P0.01），细胞凋亡被显著抑制(凋亡率从13.50%降为2.08%，t=7.90，P0.01)，细胞分裂活性增强（G2期+S期细胞比例从28.54%升为54.05%，t=6.32,P0.01）。 结论 人眼内PEDF可能作为LEC营养因子发挥细胞周期调控和抗凋亡的重要作用，参与晶状体生理性状维持和白内障发生发展。 关键词：色素上皮衍生因子（PEDF），晶状体上皮细胞（LEC）/人，房水，体外培养 Effects of PEDF on human lens epithelial cells growth regulation in vivo and in vitro LIU Tian1，XIANG Dao-man1，JIANG Fu-neng2 1.Department of Ophthalmology，Guangzhou Women and Children’s Medical Center，Guangzhou 510623； 2.Guangdong?Key?Laboratory?of?Clinical?Molecular?Medicine?and?Diagnostics，Guangzhou?First?People’s?Hospital，Guangzhou?Medical?University，Guangzhou 510180 Abstract Purpose：To investigate in vivo and in vitro the role of PEDF on human lens epithelial cells biological behaviour regulation. Methods：In vivo study，aqueous PEDF concentration was detected by enzyme-linked immunosorbent assay(ELISA) in age-related cataract patients who underwent cataract phacoemulsification extraction surgery. The relationship of PEDF level was analyzed with LECs?density on anterior lens capsular specimens and?cataract?types. In vitro study,PEDF was added in culture media of human lens epithelial cell line?(HLE-B3). The cells growth?activity was detected by Cell Counting Kit(CCK8). Flow cytometry with AnnexinV-FITC/7-AAD double staining method was used to detect the changes of cells cycle and apoptosis. Results：The mean aqueous PEDF concentration of 120 cataract cases was 156.2?± 41.2ng/ml. The central?anterior subcapsular LEC density showed signifi