[Effects of hypoxic preconditioning on glucose metabolism of rat bone marrow mesenchymal stem cells]

To study the effects of hypoxic preconditioning on the glucose metabolism of rat BMSCs and its underlying mechanism so as to provide the theoretical basis for the optimization of the stem-cell based therapy.

Density gradient centrifugation method was adopted to isolate rat BMSCs from neonatal SD rats (aged 1-3 days). BMSCs were cultured to 4th passage and divided into 4 groups based on different culture conditions: group A in normoxia condition for 24 hours, group B in 1% O2 for 24 hours, group C in 2-methoxyestradiol (20 micromol/L) for 24 hours before hypoxic preconditioning, and group D in hypoxia-inducible factor 1 (HIF-1) specific siRNA (50 micromol/L) for 12 hours before hypoxic preconditioning. MTT method was applied to evaluate the proliferation of BMSCs. Biochemical analyzer and Real-time fluorescent quantitative PCR were applied to detect the glucose uptake, lactate production, and HIF-1alpha mRNA and Glut-1 mRNA levels of BMSCs.

MTT showed that the absorbance (A) values were 387.67 +/- 58.92, 322.50 +/- 50.60, 297.00 +/- 53.00, and 286.00 +/- 41.00 in groups A, B, C, and D, respectively, showing no significant difference among 4 groups (P > 0.05). The levels of glucose uptake and lactate production were higher in group B than in groups A, C, and D, showing significant differences (P < 0.05); the levels of groups C and D were higher than those of group A, but showing no significant difference (P > 0.05). The mRNA expressions of HIF-1alpha and Glut-1 elevated significantly in group B when compared with those in group A (P < 0.05); groups C and D were significantly lower than group B (P < 0.05) and were significantly higher than group A (P < 0.05).

Hypoxic preconditioning can stimulate the glucose uptake and metabolism of rat BMSCs, whose mechanism is probably related to up-regulating the mRNA expressions of HIF-1alpha and Glut-1.