目的：阐明在EGCG抗兔晶状体上皮细胞增殖中PKB通路的作用。
方法：（1）兔晶状体上皮细胞增殖抑制率的测定。应用 MTT法测定用药后及加用特异性的PKB活性的抑制剂LY294002后，晶状体上皮细胞的增殖抑制率。（2）用形态学及琼脂糖凝胶电泳检测兔晶状体上皮细胞的凋亡，（3）应用WESTERN BLOT法研究EGCG对PKB系统的蛋白激酶磷酸化和非磷酸化的水平的影响。
结果：（1）EGCG对晶状体上皮细胞的增殖抑制率有明显的量效和时效关系；（2）EGCG可以诱导晶状体上皮细胞的凋亡；（3）EGCG通过抑制PKB的磷酸化水平而抑制晶状体上皮细胞的增殖；EGCG对PKB活性的影响主要在于调节蛋白激酶的磷酸化与去磷酸化，对其总蛋白含量影响不大。
结论：EGCG通过诱导晶状体上皮细胞的凋亡和抑制PKB的磷酸化水平而抑制晶状体上皮细胞的增殖。
Purpose: Study the role of PKB signal transduct pathway in the antiproliferation effect induced by EGCG.
Methods: (1) Cell proliferation inhibition rate was examinated by MTT colormetric assay;(2) Cell apoptosis was detected morphology and agarose gel electrophoresis. (3) Western blotting were used to study the effect of EGCG on the kinsae phosphorylation- and nonphosphorylation- level of PKB.
Result: (1) EGCG could inhibt rabbit LECs in an time- and dose-depent manner;(2) EGCG could induce rabbit LECs apoptosis; (3) Both basic phosphorylation- and nonphosphorylation- level of PKB were fairy high in LECs, but the phosphorylation- level decreased with EGCG concentration increased. The nonphosphorylation- level of PKB remained constant in all test. 15min after 200μmol/L was added, the phosphorylation- level was the highest and began to wane.
Conclusions: EGCG could inhibit LECs proliferation by inducing apoptosis and lowing PKB phosphorylation- level.