摘要
目的观察脯氨酸羟化酶抑制剂二甲基乙二酰基甘氨酸(dimethyloxalglycine,DMOG)对无血清培养引起的小鼠骨髓间充质干细胞的凋亡影响及其对凋亡调控蛋白Bcl-2/Bax表达的影响。方法分离小鼠股骨及胫骨骨髓腔中的单个核细胞,传代后应用流式细胞仪进行细胞表型检测和三系分化以鉴定骨髓间充质干细胞。流式细胞仪TUNEL法观察DMOG作用于细胞后对细胞无血清培养凋亡的对抗作用,Westerm blot检测DMOG对凋亡调控蛋白Bcl-2/Bax蛋白表达的影响。结果实验所获的单个核细胞通过流式细胞仪细胞表型鉴定和三系分化鉴定证实为骨髓间充质干细胞。与无血清造模组相比,二甲基乙二酰基甘氨酸能明显降低骨髓间充质干细胞因缺血清导致的凋亡率(P<0.05),并上调凋亡抑制蛋白Bcl-2蛋白表达(P<0.05),抑制凋亡促进蛋白Bax蛋白表达(P<0.05)。结论脯氨酸羟化酶抑制剂DMOG能抑制骨髓间充质干细胞因缺血清培养引起的凋亡。对凋亡调控蛋白Bcl-2/Bax表达的调控可能是DMOG对抗骨髓间充质干细胞凋亡的重要机制。
Objective To evaluate the effect of prolyl hydroxylase inhibitor, dimethyloxalglycine （DMOG） , on the apoptosis of mouse bone marrow mesenchymal stem cells （BMSCs） induced by serum deprivation and the expression of apoptosis regulation proteins, Bcl-2/Bax. Methods Mononuclear cells in the cavity of the femur and tibia in mice were isolated. After passaging, cell phenotype was identified using flow cytometry. BMSCs were identified using tri-lineage differentiation. The effect of DOMG on the apoptosis induced by serum deprivation was assessed using TUNEL staining and flow cytometry. The expression of Bcl-2/Bax was evaluated with Western blotting. Results The mononuclear cells isolated in this experiment were identified as BMSCs using flow cytometry for cell phenotype and tri-lineage identification. Compared to serum-free modeling group, DMOG could obviously reduce the apoptosis rate of BMSCs induced by serum deprivation （P 〈 0. 05） , up-regulate the expression of Bcl-2 protein （P 〈 0. 05 ） , and down-regulate the expression of Bax protein （P 〈 0. 05）. Conclusion Prolyl hydroxylase inhibitor DMOG can inhibit the apoptosis of BMSCs induced by serum deprivation. The regulation of the expression of Bcl-2/Bax may be an important mechanism of the anti-apoptotie effect of DMOG on BMSCs.
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2013年第1期21-25,共5页
Chinese Journal of Osteoporosis
基金
云南省卫生厅科研基金(2011WS0033)
