EGFR和Ca2+在外源性EGF刺激大鼠A型精原细胞增殖中的作用

摘要

目的 探究表皮生长因子受体(EGFR)和Ca2+在表皮生长因子(EGF)引起的大鼠A型精原细胞增殖中的作用.方法 选用9 d龄的SD大鼠,使用Percoll不连续密度梯度分离纯化精原干细胞;实验一:将细胞分为对照组、AG1478组、EGF组和EGF+AG1478组,观察EGFR在EGF促A型精原细胞增殖中的作用;实验二:将细胞分为对照组、Ca2+鳌合剂(BAPTA)组、EGF组、EGF+BAPTA组,观察细胞内游离Ca2+在EGF促A型精原细胞增殖中的作用;实验三:将细胞分为对照组、EGF组和EGF+AG1478组,采用Fura-2/AM负载法检测细胞内游离的Ca2+浓度.结果 外源性的EGF明显促进了A型精原细胞的增殖;AG1478和BAPTA明显减弱了EGF对细胞的促增殖作用.结论 EGF通过与EGFR特异性结合提高胞内Ca2+浓度对A型精原细胞增殖起促进作用.%Objective To study the effects of EGFR and Ca + on the proliferation of type A spematogonial cells by epidermal growth factor ( EGF ). Methods Type A spermatogonial cells were isolated from 9 day-old SD rat testes by discontinuous percoll gradient centrifugation. Experiment 1 : cells were divided into control group, AG1478 group, EGF group and EGF + AG1478 group, to observe EGFR mediated effect of EGF on proliferation of type A spermatogonial cells. Experiment 2:cells were divided into control group, BAPTA group, EGF group and EGF + BAPTA group,to observe Ca + mediated effect of EGF on proliferation of type A spermatogonial cells. Experiment 3 included control group, EGF group and EGF + AG1478 group. The intracellular free Calcium concentration was measured by using Fura-2/AM as a fluorescent indicator. Results Exogenous EGF apparently stimulated the proliferation of type A spematogonial cells. Effects of EGF on proliferation of type A spematogonial cells was reduced markedly by AG1478 and BAPTA. Conclusion EGF promotes proliferation of the type A spematogonial cells through binding specifically to EGFR and increasing the intracellular free Calcium concentration.