Adrenomedullin effect on expression of apoptosis-related protein in bone marrow mesenchymal stem cells under hypoxia

doi:10.3969/j.issn.2095-4344.2015.32.002

Abstract

Abstract:

BACKGROUND: Adrenomedullin gene transfection can strength the anti-apoptotic ability of bone marrow mesenchymal stem cells under ischemia and hypoxia, but its mechanism is not yet clear.
OBJECTIVE: To investigate the effect of adrenomedullin on the expression of apoptosis-related proteins, Bax, Bcl-2 and Caspase-3, in bone marrow mesenchymal stem cells under hypoxia and ischemia.
METHODS: Bone marrow mesenchymal stem cells of Sprague-Dawley rats were isolated, cultured and purified, and then cultured in serum-free medium under hypoxic condition for 0, 3, 6, 9, 12 hours. Then, western blot assay was employed to detect the expression of Bax, Bcl-2 and Caspase-3 so as to determine the optimal hypoxia time that was determined at 6 hours of hypoxia. Depending on whether adrenomedullin pretreatment was done, the cells were divided into control group (with no adrenomedullin pretreatment before hypoxia and ischemia) and adrenomedullin groups with different concentrations (1, 10, 100 μg/L). Afterwards, the expression of Bax, Bcl-2 and Caspase-3 was detected by using western blot assay.
RESULTS AND CONCLUSION: (1) After cultured in serum-free medium under hypoxia for 0, 3, 6, 9, 12 hours, the expression of Bax, Bcl-2 and Caspase-3 in bone marrow mesenchymal stem cells were increased (P < 0.05);
at 6 hours of hypoxia, the Bax/Bcl-2 ratio and Caspase-3 expression reached the minimum value (P < 0.05). (2) At 6 hours of hypoxia, the expression of Bax and Caspase-3 protein as well as Bax/Bcl-2 ratio became the lowest in the
100 μg/L group compared with the 1 and 10 μg/L groups, but the expression of Bcl-2 protein reached the peak (P < 0.05). These findings indicate that adrenomedullin can reduce the expression of Bax/Bcl-2 ratio and Caspase-3 protein in bone marrow mesenchymal stem cells cultured in serum-free medium under hypoxic conditions, which is in a dose-dependent manner.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cells, Adrenomedullin, Apoptosis Regulatory Proteins

CLC Number:

R394.2

Yao Yue-xian, Wang Jun-sheng, Su Jin-zi, Jiang Jin-feng, Cai Wen-qin. Adrenomedullin effect on expression of apoptosis-related protein in bone marrow mesenchymal stem cells under hypoxia[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(32): 5092-5096.

Figures/Tables 2

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