%0 Journal Article
%T PDGFR信号通路促进星形胶质细胞和成纤维细胞共培养时细胞簇的成团效应<br>PDGFR signaling pathway promotes cluster formation in co-culture system of astrocytes and fibroblasts
%A 裴 丹
%A 李洪鹏
%J 西安交通大学学报(医学版)
%D 2019
%R 10.7652/jdyxb201904015
%X 摘要：目的 探讨血小板衍生生长因子受体(PDGFR)信号通路对小鼠脑星形胶质细胞和成纤维细胞共培养体系增殖的影响。方法 取出生1～3d的小鼠乳鼠大脑皮质培养星形胶质细胞，取脑膜培养成纤维细胞，免疫细胞化学荧光染色胶质纤维酸性蛋白(GFAP)鉴定星形胶质细胞纯度，染色纤连蛋白(FN) 鉴定成纤维细胞纯度。通过向小鼠脑星形胶质细胞和成纤维细胞共培养细胞体系中加入细胞因子血小板衍生生长因子-BB(PDGF-BB)刺激细胞或抑制剂AG1296进行干预，观察各组细胞增殖情况，Western blot检测共培养细胞系中p-PDGFR-β、磷酸化的蛋白激酶B(p-Akt)的表达变化。结果 鉴定结果显示星形胶质细胞和成纤维细胞的细胞纯度达到90%。显微镜下观察显示，加入PDGF-BB后星形胶质细胞和成纤维细胞共培养体系增殖明显并形成团块，给予抑制剂AG1296则增殖受到抑制，团块明显减少。Western blot结果显示加入PDGF-BB时，p-PDGFRβ、p-Akt表达增多(P<0.01)；给予抑制剂AG1296则表达减少(P<0.05)。结论 PDGFR信号通路促进体外小鼠脑星形胶质细胞和成纤维细胞的增殖。<br>ABSTRACT: Objective To investigate the effects of platelet-derived growth factor receptor (PDGFR) signaling pathway on the proliferation of mouse brain astrocytes and fibroblasts co-culture system. Methods Astrocytes were cultured in the cerebral cortex of mice born 1-3 days. The meninges were cultured to form fibroblasts. Immunocytochemical fluorescence staining of glial fibrillary acidic protein (GFAP) was used to identify the purity of the cultured astrocytes. Fibronectin (FN) was used to identify the purity of the fibroblasts. By adding platelet-derived growth factor BB (PDGF-BB) to the co-cultured cell system of mouse brain astrocytes and fibroblasts and intervening with inhibitor AG1296, we detected the proliferation of cells treated with different methods. We also detected the expressions of p-PDGFR-β and phosphorylated protein kinase B (p-Akt) in co-cultured cell system by Western blot. Results The purification and identification of the astrocytes and fibroblasts showed that the purity of the cells reached 90%. Microscopic observation showed that the proliferation of the astrocytes and fibroblasts co-cultured was obvious and clusters were formed when PDGF-BB was added. The proliferation of cell clusters was significantly reduced after AG1296 was administered. Western blot showed that the expression of p-PDGFRβ and p-Akt increased after PDGF-BB was added (P<0.05), and decreased after administration of AG1296 (P<0.05). Conclusion PDGFR signaling pathway promotes the proliferation of astrocytes and fibroblasts in vitro
%K 星形胶质细胞
%K 成纤维细胞
%K 血小板衍生生长因子
%K 血小板衍生生长因子受体
%K 细胞增殖<br>astrocyte
%K fibroblast
%K PDGF-BB
%K PDGFR
%K cell proliferation
%U http://yxxb.xjtu.edu.cn//oa/darticle.aspx?type=view&id=201904015