Effects of glucose on human bone marrow mesenchymal stem cell growth, apoptosis and senescence

English Keywords:Mesenchymal stem cells  Glucose  Cell growth  Cells apoptosis  Cell senescence

English Abstract:

  【Abstract】 Objectives: To investigate the effect on cell growth, cycle, apoptosis and senescence of human bone marrow mesenchymal stem cells in high concentration of glucose microenvironment or with high level of O-GlcNAcylation cell models, and to explore the underlying molecular mechanism. Methods: The extent of growth of hMSCs previously cultured in normal concentration of glucose(group A)(1.0mg/ml), high concentration of glucose(group B)(4.5mg/ml), normal glucose with thiamet-G(group C)(1.0mmol/L) and high glucose with thiamet-G(group D)(1.0mmol/L) was evaluated by WST-1, flow cytometer, β-galactosidase staining to identify the role of glucose and O-GlcNAcylation in the regulation of proliferation, cell cycle, apoptosis and senescence in hMSCs. In addition, the expressions of Cyclin D1 and caspase-3 were detected by real-time RT-PCR and western blot. Results: In group B, group C and group D, hMSCs grew fewer than which in group A on the third, fifth and seventh day(P<0.05). Also in group B, group C and group D, the induced cell cycle was arrested at G1 phase in hMSCs on the fifth day(P<0.05). The rate of apoptosis of hMSCs in group B, group C and group D was higher than that in group A(P<0.05). After treatment, the dimension of hMSCs in group B, group C and group D was larger than that in group A culture(P<0.05), and the positive cells of SA-β-gal staining were also more than those in group A on the seventh day(P<0.05). The real-time PCR showed thawt the expression of Cyclin D1 in group A, B, C and D was 1.01±0.31, 0.31±0.07, 0.42±0.1, 0.18±0.04, and it decreased in group A on the fifth day(P<0.05). The expression of aspase-3 in group A, B, C and D was 1.09±0.82, 5.73±1.54, 3.43±0.59, 6.82±2.13, and it increased in group A on the fifth day(P<0.05). Compared with group A, the expression of Cyclin D1 decreased and the expression of aspase-3 increased when hMSCs were cultured in group B, group C and group D on the fifth day(P<0.05). Conclusions: Our findings demonstrate the effect of glucose concentration and O-GlcNAcylation on regulating the growth, cycle, apoptosis and senescence of hMSCs, which provide insight into the mechanism of how glucose concentration regulating hMSCs.