摘要
背景:研究表明体外培养细胞加入胸腺素β4能增加细胞的抗凋亡能力,若在缺氧环境中上调胸腺素β4基因在骨髓间充干细胞中的表达,其抗凋亡能力如何改变,目前鲜见报道。目的:观察胸腺素β4基因修饰的骨髓间充质干细胞在缺氧环境中的凋亡率改变,并探讨其是否通过调控Bax、Bcl-2表达影响凋亡能力。方法:将携带有胸腺素β4基因的慢病毒转染骨髓间充质干细胞,转染完毕后利用Western blot法检测胸腺素β4在骨髓间充质干细胞中的表达。将细胞分为胸腺素β4转染组、对照病毒组、未转染组,分别置于缺氧环境中。流式细胞仪检测3组细胞凋亡率;Western blot法检测3组细胞中Bax和Bcl-2蛋白表达情况。结果与结论:Western blot检测结果示,胸腺素β4基因在骨髓间充质干细胞中成功表达。流式细胞仪结果示,胸腺素β4转染组细胞凋亡率较对照病毒组、未转染组低,而对照病毒组、未转染组凋亡率差异无显著性意义。Western blot结果显示,Bcl-2蛋白在胸腺素β4转染组中表达量较对照病毒组、未转染组高,Bax蛋白在胸腺素β4转染组表达量较对照病毒组、未转染组低,而对照病毒组、未转染组中Bax、Bcl-2表达差异无显著性意义。提示过表达胸腺素β4基因可增加骨髓间充质干细胞在缺氧环境中的抗凋亡能力,其可能的作用机制是调控Bax和Bcl-2蛋白表达水平。
BACKGROUND：Studies have shown that thymosinβ4 can improve the anti-apoptotic ability of a variety of cells, but the reports describing the changes in the anti-apoptotic ability of bone mesenchymal stem cells modified with thymosinβ4 gene in the hypoxic environment are rare.OBJECTIVE：To observe the change in the apoptotic rate of bone mesenchymal stem cells modified with thymosinβ4 gene in the hypoxic environment, and to explore whether thymosinβ4 affects the apoptosis of bone marrow mesenchymal stem cells via regulating the expression of Bax and Bcl-2.METHODS：Bone marrow mesenchymal stem cells were transfected with the recombinant lentiviral vector over-expressing the thymosinβ4 gene, and then we observed the expression of thymosinβ4 using western blot assay. cells were divided into three groups：thymosinβ4 transfection group, control virus group, and untreated group. Al three groups were placed in a hypoxic environment. The apoptotic rate of bone marrow mesenchymal stem cells was evaluated by the flow cytometry assay. The expression of Bax and Bcl-2 protein was detected by western blot.RESULTS AND CONCLUSION：Thymosinβ4 gene was expressed successful y in the bone marrow mesenchymal stem cells showed by the western blot. The apoptotic rate of bone marrow mesenchymal stem cells in the hypoxic environment was lower in the thymosinβ4 transfection group than the control virus group and untreated group;while there was no difference between the latter two groups. Western blot results showed that the expression of Bcl-2 protein was higher in the thymosinβ4 transfection group than the control virus group and untreated group, and the expression of Bax protein was lower in the thymosinβ4 transfection group than the control virus group and untreated group. No difference in the expression of Bax and Bcl-2 was found between the control virus group and untreated group. These findings indicate that thymosinβ4 overexpression can improve the anti-apoptotic ability of bone mesenchymal stem cells modified in the hypoxic environment, and its possible mechanism is through regulating the expression of Bax and Bcl-2 protein.
出处
《中国组织工程研究》
CAS
CSCD
2014年第41期6573-6577,共5页
Chinese Journal of Tissue Engineering Research
基金
武汉晨光计划项目(200950431177)~~
