Abstract

Objective To observe the effects of liraglutide on proliferation and apoptosis of human umbilical cord mesenchymal stem cells (HUC-MSCs) in vitro, and further to investigate the relationship with phosphatidylinositol 3 kinase/serine-threonine kinase (PI3K/AKT) signaling pathway. Methods HUC-MSCs cultured in DMEM/F12 media were treated with varying concentrations of liraglutide(10-9, 10-8, 10-7 mol/L and control) for 24 and 48 hours separately. In order to explore the best concentrations and application time, cell proliferation was detected by cell counting kit-8 (CCK-8) assay, apoptosis rate was detected by flow cytometry. LY294002, the specific inhibitor of PI3K, was added in the culture at 20 μmol/L concentration. The cyclic adenosine monophosphate (cAMP) concentration was detected by ELISA, the protein expression of serine-threonine kinase (AKT) and phosphorylated AKT (P-AKT) were measured by Western blotting. Difference between two groups was analyzed by t test, and variance analysis was used for multiple variables analysis. Results Compared with 10-9 mol/L group (0.52±0.10) , 10-8 mol/L group (0.68±0.07) and control group (0.44±0.09) separately, cell proliferation was increased significantly in 10-7 mol/L liraglutide group (0.78±0.04) after 24 hours (t=8.383, 4.167, 12.100, all P<0.05). Compared with 10-9 mol/L group (3.295±0.346) and control group (16.315±0.474) , apoptosis rate was decreased significantly in 10-7 mol/L liraglutide group (1.380±0.255) (t=6.299, 39.272, all P<0.05); Compared with control group and LY294002 group separately, the concentration of cAMPwas increased significantly in liraglutide group (t= 14.307, 23.378, all P<0.05) , and the expression of p-AKT/AKT was increased in liraglutide group (t=7.861, 8.486, all P<0.05); Compared with liraglutide group, the concentration of cAMP and the expression of p-AKT/AKT were all decreased in liraglutide + LY294002 group(t=16.473, 3.172, all P<0.05). Conclusions Liraglutide can promote proliferation and inhibit apoptosis of HUC-MSCs and the optimal concentration and culture time are 10-7 mol/L and 24 hours. The mechanism may be related to the activation of the PI3K/AKT signaling pathway. Key words: Liraglutide; Human umbilical cord mesenchymal stem cells; Proliferation; Apoptosis

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