表皮生长因子影响大鼠骨髓间充质干细胞增殖和迁移的信号通路

发布时间：2018-04-03 18:28

本文选题：表皮生长因子　切入点：间质干细胞　出处：《中国组织工程研究》2017年29期

【摘要】：背景:表皮生长因子作为一种辅助生长因子,但是对于骨髓间充质干细胞的生长是否必须尚无定论。目的:建立大鼠骨髓间充质干细胞分离提取以及鉴定的方法,同时探索表皮生长因子对其增殖能力以及迁移能力的影响,同时探索其潜在机制。方法:①采用全骨髓贴壁法,分离纯化大鼠骨髓间充质干细胞,培养至第3代,使用流式细胞仪检测细胞表面抗原CD29,CD45,CD90表达情况。通过成骨和成脂分化诱导,更进一步鉴定骨髓间充质干细胞;②同时通过CCK-8法和克隆形成实验测量表皮生长因子对骨髓间充质干细胞的第3代细胞的增殖能力的影响。同时通过Transwell小室验证表皮生长因子对骨髓间充质干细胞的第3代细胞迁移能力的影响。通过蛋白印迹实验检测PI3K/Akt和核因子κB信号通路相关蛋白表达水平,探索表皮生长因子的作用机制。结果与结论:①原代骨髓间充质干细胞呈多角形和梭形,随着培养时间增加,逐渐呈梭形为主。流式细胞术检测第3代细胞表面CD29,CD90表达呈阳性,CD45表达成阴性。并且对骨髓间充质干细胞的第3代细胞成功进行了成骨和成脂分化诱导;②表皮生长因子能够促进第3代细胞的增殖,且细胞中PI3K/Akt信号通路关键分子p-Akt表达水平的增加,同时其下游信号分子Bax表达水平降低而Bcl-2表达升高。表皮生长因子能够通过调节核因子κB信号通路分子p 65以及IκB的磷酸化水平,同时也增加了基质金属蛋白酶9的表达水平,从而促进大鼠骨髓间充质干细胞的迁移;③结果表明表皮生长因子促进骨髓间充质干细胞的增殖和迁移。
[Abstract]:Background: epidermal growth factor (EGF) is an auxiliary growth factor, but it is not clear whether the growth of bone marrow mesenchymal stem cells (BMSCs) must be done.Aim: to establish a method for isolation, extraction and identification of rat bone marrow mesenchymal stem cells (BMSCs), and to explore the effects of EGF on its proliferation and migration, as well as its potential mechanism.Methods the bone marrow mesenchymal stem cells of rats were isolated and purified by whole bone marrow adherent method, and cultured to the third passage. The expression of CD29, CD45, CD90 was detected by flow cytometry.The effects of epidermal growth factor on the proliferation of bone marrow mesenchymal stem cells (BMSCs) were measured by CCK-8 assay and clone formation assay.The effects of epidermal growth factor (EGF) on the migration ability of the third generation of bone marrow mesenchymal stem cells (BMSCs) were also tested by Transwell chamber.To investigate the mechanism of epidermal growth factor (EGF), PI3K/Akt and NF- 魏 B signaling pathway related proteins were detected by Western blotting.Results and conclusion the primary bone marrow mesenchymal stem cells were polygonal and fusiform.Flow cytometry was used to detect the positive expression of CD29 + CD90 and negative expression of CD45 on the surface of the third passage cells.Furthermore, osteogenic and adipogenic differentiation induced by EGF 2 in the third passage of bone marrow mesenchymal stem cells could promote the proliferation of the third passage cells, and the expression of p-Akt, a key molecule of the PI3K/Akt signaling pathway, was increased in the cells.At the same time, its downstream signal molecule Bax expression level decreased, but the Bcl-2 expression increased.Epidermal growth factor can promote the migration of rat bone marrow mesenchymal stem cells by regulating the phosphorylation level of nuclear factor- 魏 B signaling pathway p65 and I 魏 B and increasing the expression of matrix metalloproteinase-9.The results showed that epidermal growth factor promoted the proliferation and migration of bone marrow mesenchymal stem cells.
【作者单位】：遵义医学院生物化学与分子生物学教研室;遵义医学院附属医院胸外科;遵义医学院附属医院耳鼻喉科;
【基金】：贵州省科学技术基金(黔科合J字[2012]2360号)~~
【分类号】：R329.2

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