Basic FGF Promotes Proliferation of Ovarian Granulosa Cells in the Laying Chickens Via FGFR1 and PKC Pathway
Basic FGF Promotes Proliferation of Ovarian Granulosa Cells in the Laying Chickens Via FGFR1 and PKC Pathway
复制标题
碱性 FGF 通过 FGFR1 和 PKC 途径促进蛋鸡卵巢颗粒细胞增殖
DOI：
10.1111/j.1439-0531.2011.01813.x
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发表时间：
2012-02-01
影响因子：
1.7
通讯作者：
Zhang, C.
中科院分区：
文献类型：
--
作者：
Lin, J.;Jia, Y.;Zhang, C.
研究了碱性成纤维细胞生长因子对蛋鸡卵巢分级前卵泡颗粒细胞的促增殖作用。用免疫组织化学和RT-PCR方法检测小黄囊中碱性成纤维细胞生长因子受体1的表达。FGFR1蛋白和信使核糖核酸主要在颗粒层表达。碱性成纤维细胞生长因子(0.1100 ng/ml)作用8~24 h后，颗粒细胞增殖明显增强，并呈剂量和时间依赖性。FGFR1拮抗剂SU5402可抑制bFGF诱导的细胞增殖。5-溴-2-脱氧尿苷掺入和末端转移酶dUTP缺口末端标记实验进一步证实了这种刺激作用。蛋白激酶A(PKA)和蛋白激酶C(PKC)的免疫细胞化学结果显示，碱性成纤维细胞生长因子的促增殖作用主要激活PKC的表达。同时，PKC激活剂PMA可显著促进bFGF诱导的细胞增殖，而PKC抑制剂H7则可抑制其增殖(p<；0.05)。此外，bFGF诱导的卵巢颗粒细胞增殖还伴随着细胞周期调控基因细胞周期蛋白D1和E1、细胞周期蛋白依赖性蛋白激酶2和细胞周期蛋白依赖蛋白6基因表达的增加。结论：bFGF通过与FGFR1结合并参与产蛋鸡分级前卵泡的PKC通路，促进了卵巢颗粒细胞的增殖。
The proliferating effect of basic fibroblast growth factor (bFGF) on granulosa cells from ovarian pre-hierarchical follicles was evaluated in the laying chickens. Expression of bFGF receptor 1 (FGFR1) from small yellow follicles was determined by immunohistochemistry and RT-PCR. The FGFR1 protein and mRNA were intensively expressed in the granulosa layer. After 8- to 24-h treatment with bFGF (0.1100 ng/ml), the proliferation of cultured granulosa cells was remarkably enhanced in a dose- and time-dependent manner. The FGFR1 antagonist SU5402 inhibited bFGF-induced cell proliferation. This stimulating effect was further confirmed by 5-bromo-2-deoxyuridine incorporation and terminal transferase dUTP nick end-labelling assay. Immunocytochemistry of protein kinases A (PKA) and C (PKC) showed that the pro-proliferation action of bFGF predominantly activated PKC expression. Meanwhile, the bFGF-induced cell proliferation was significantly promoted by PKC activator PMA and inhibited by PKC inhibitor H7 (p < 0.05). In addition, the bFGF-elicited cell proliferation was accompanied with increased mRNA expression of the cell cycle-regulating genes including cyclins D1 and E1, cyclin-dependent kinases 2 and 6. In conclusion, bFGF promoted the proliferation of ovarian granulosa cells through binding with FGFR1 and involving PKC pathway in the pre-hierarchical follicles of the laying chickens.