周维维,李豆豆,张洁云,刘思麟,李春绒,曹 猛.Periostin表达上调对去势大鼠骨髓间充质干细胞生物学行为的作用[J].,2019,19(10):1864-1869 |
Periostin表达上调对去势大鼠骨髓间充质干细胞生物学行为的作用 |
The Effect of Periostin Expression up-regulation on the Biological Behaviors of Bone Marrow Mesenchymal Stem Cells in Ovariectomized Rats |
投稿时间:2019-01-22 修订日期:2019-02-27 |
DOI:10.13241/j.cnki.pmb.2019.10.012 |
中文关键词: Periostin 绝经后骨质疏松症 骨髓间充质干细胞 成骨分化 |
英文关键词: Periostin Postmenopausal osteoporosis BMSCs Osteoblastic differentiation |
基金项目:国家自然科学基金面上项目(81470774) |
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中文摘要: |
摘要 目的:探究Periostin(骨膜蛋白)表达上调对雌性去势大鼠骨髓间充质干细胞(BMSCs)成骨分化、细胞增殖与凋亡特性的作用。方法:通过去势手术建立雌性大鼠骨质疏松模型,待建模成功后分离培养并鉴定BMSCs,利用含有增强型绿色荧光蛋白(EGFP)和大鼠Periostin基因的重组慢病毒转染P3代BMSCs,成骨诱导后鉴定其成骨分化能力改变,流式细胞仪检测其细胞周期以及细胞凋亡率的变化。结果:成功建立骨质疏松模型;荧光显微镜下观察到绿色荧光提示慢病毒载体实现转染并表达目的蛋白;慢病毒转染组BMSCs成骨诱导后ALP及茜素红染色较去势组BMSCs染色加深;慢病毒转染组BMSCs的S期细胞比例为(17.07±0.56)%,显著高于去势组BMSCs的S期细胞比例(8.42±0.02)%,差异具有统计学意义(P<0.05);慢病毒转染组BMSCs的细胞凋亡率为(7.3±0.1)%,显著低于去势组BMSCs的凋亡率(12.05±0.55)%,其差异具有统计学意义(P<0.05)。结论:Periostin表达上调可提高去势骨髓间充质干细胞的成骨分化及细胞增殖能力,并对其凋亡有抑制作用。 |
英文摘要: |
ABSTRACT Objective: To explore the effects of Periostin up-regulation on osteoblastic differentiation, cell proliferation and apoptosis of bone marrow mesenchymal stem cells (BMSCs) in ovariectomized rats. Methods: An ovariectomized animal model was established by castrated surgery. Then BMSCs were separated, cultured and identified. BMSCs of P3 generation were transfected by recombinant lentivirus which have enhanced green fluorescent protein (EGFP) and rat Periostin gene. The osteoblastic differentiation ability were determined after osteoblastic induction. The cell cycle and cell apoptosis of BMSCs were detected by means of flow cytometry instrument. Results: The ovariectomized animal model was established successfully. Green fluorescence under fluorescence microscope indicated that lentivirus vector had been transfected and expressed the target protein. ALP and alizarin red staining of BMSCs in lentivirus transfection group after osteoblastic induction were deeper than that in castrated group. The proportion of s-phase cells in lentivirus transfected group was (17.07 0.56) %, significantly higher than that in castrated group (8.42 0.02) %, the difference was statistically significant (P < 0.05). The apoptosis rate of BMSCs in lentivirus transfected group was (7.3 0.1) %, significantly lower than that in casted-group (12.05 0.55) %, and the difference was statistically significant (P < 0.05). Conclusion: Periostin up-regulation could improve osteoblastic differentiation and proliferation of bone marrow mesenchymal stem cells (BMSCs), and inhibit apoptosis. |
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