层黏连蛋白促进人晶状体上皮细胞蛋白激酶B的活性  

Laminin increase protein kinase B activity of human lens epithelial cells

作　　者：王欣玲[1] 阎启昌[1] 张国刚[2] 于韬[3] 张劲松[1] 

机构地区：[1]中国医科大学附属第四医院眼科,中国辽宁省沈阳市1110005 [2]沈阳药科大学中药学院,中国辽宁省沈阳市2110016 [3]辽宁省肿瘤医院医学影像科,中国辽宁省沈阳市3110042

出　　处：《国际眼科杂志》2006年第5期1018-1020,共3页International Eye Science

基　　金：中国国务院第33批博士后科学基金资助项目(No.20030033282);中国辽宁省教育厅高等学校科学研究项目(No.2004C041);中国辽宁省教育厅高等学校科学技术研究项目(No.05L576)~~

摘　　要：目的:探讨层黏连蛋白(Ln)对传代培养的人晶状体上皮细胞(hLECs)的蛋白激酶B(PKB)活性的影响,以及Ln激活PKB的信号分子途径。方法:选取第5代hLECs,分为对照组和处理组,在处理组的培养液中加入层黏连蛋白,终浓度为5mg/L。分别在处理后0,10,20,40,60min取出细胞,[γ-32P]-ATP掺入法测定胞膜和胞质PKB比活性。以PI3K的特异性抑制剂Wortmannin(终浓度100nmol/L)预处理1h,再加入层黏连蛋白,0,10,20,40,60min测定胞膜和胞质PKB比活性。结果:hLECs胞膜和胞质PKB比活性在层黏连蛋白作用40min时达到最高值,分别为空白对照的2.72和2.00倍。层黏连蛋白处理的各组胞膜和胞质PKB比活性显著高于对照组(P<0.05)。经Wortmannin预处理后,再加入层黏连蛋白,胞膜和胞质PKB比活性与未经预处理的各时间组相比均显著下降(P<0.05)。结论:层黏连蛋白通过PI3K途径激活人晶状体上皮细胞的PKB活性,比活性随时间变化而发生改变。AIM： To investigate the effects of Laminin （Ln） on protein kinase B （PKB） of the human lens epithelial cells （hLECs） subcultured in vitro and the laminin-activated PKB signaling pathway. METHODS： The fifth passage hLECs were seeded in culture bottles belong to control or treated group. Laminin was added into the nutrient medium of the treated group with the end concentration of 5μg/mL. The cells were collected for 0, 10, 20, 40 and 60 minutes after Laminin treatment, respectively. PKB Kinase Assay [γ -32P]-ATP incorporation assay was used to detect the PKB kinase activity. Wortmannin, the specific inhibitor of phosphatidylinosital-3-kinase （PI3K）, was applied to pretreat the hLECs for 1 hour at the concentration of 100nmol/L. Then Laminin was added and PKB activity was detected. RESULTS： PKB activity of the membrane and cytosol reached the peak value at 40 minutes after Laminin treat- ment, as 2.72 and 2.00 times as much as that of the control group. There was significant difference in PKB activity between the Laminin treated group and the control group. Both membrane and cytosol activity of PKB decreased significantly after Wortmannin pretreatment. CONCLUSION： Laminin activate both membrane and cy- tosol PKB activity through the PI3K pathway and the PKB activity varied according to the time.

关 键 词：层黏连蛋白 晶状体 上皮细胞 蛋白激酶B 

分 类 号：R776[医药卫生—眼科]