Role of AMD3100 combined with granulocyte colony-stimulating factor in the mobilization of bone marrow endothelial progenitor cells in diabetic mice

Abstract:

Objective：To investigate the role of AMD3100 and granulocyte colony-stimulating（G-CSF） used alone or in different se－quences in the mobilization of bone marrow endothelial progenitor cells（EPCs） in diabetic mice（db/db mice），and to provide thoughts for collecting EPCs from diabetic patients for clinical treatment. Methods：With reference to the dose and methods described in related articles in China and foreign countries，AMD3100 and recombinant human granulocyte colony-stimulating（rhG-CSF） injection were used for the mobilization of bone barrow EPCs. A total of 80 male db/db mice aged 5-6 weeks were randomly divided into group A （AMD3100+NS mobilization group），group B（AMD3100+G-CSF mobilization group），group C（G-CSF+AMD3100 mobilization group），and group D （NS blank control group）. A total of 80 db/+ male mice were selected as corresponding controls and were divided into groups A’，B’，C’，and D’. Peripheral blood samples were collected from the heart on days 3，7，10，and 14 after mo－bilization，and flow cytometry was used to measure the proportion of CD34+/CD133+/Flk-1+ cells. The results were expressed as mean±standard deviation（%），and comparison and analysis were per－formed after statistical analysis. Results：On days 3，7，10，and 14 after mobilization，there was no significant difference in the proportion of CD34+/CD133+/Flk-1+ cells between groups D and D’（P>0.05），but group D’ had a higher proportion than group D（P=0.003）. In group A，the proportion of CD34+/CD133+/Flk-1+ cells gradually increased and reached the highest value on day 7 and then gradually decreased；in groups B and C，the proportion of CD34+/CD133+/Flk-1+ cells gradually increased and reached the highest value on day 10 and then gradually decreased；the proportion of CD34+/CD133+/Flk-1+ cells at the peak was significantly higher than that at the other three time points（P=0.000）. On day 10，groups B and C had a significantly higher proportion of CD34+/CD133+/Flk-1+ cells in peripheral blood of db/db mice than groups A and D（P=0.000），but there was no significant difference between groups B and C（P>0.05）. In groups A’，B’， and C’ after mobilization，the proportion of CD34+/CD133+/Flk-1+ cells gradually increased and reached the peak on day 7 and then gradually decreased；the proportion of CD34+/CD133+/Flk-1+ cells on day 7 was significantly higher than that at the other three time points（P=0.000）. On day 7，the C’ group had a significantly higher proportion of CD34+/CD133+/Flk-1+ cells than groups A’，B’， and D’（P=0.000）. Conclusion：AMD3100 combined with G-CSF has a better effect in the mobilization of bone marrow EPCs than AMD3100 alone. In non-diabetic conditions，G-CSF followed by the addition of AMD3100 has a better mobilization effect，but in diabetic conditions，AMD3100 followed by the addition of G-CSF has a better mobilization effect.