ABSTRACT
AIM:To investigate the role of prostaglandin E2receptor 2 agonist (EP2A) in proliferation and homing of human CD34 +cells. METHODS:Bone marrow fluid and peripheral blood containing stem cells were collected from healthy donors mobilized by granulocyte colony-stimulating factor in our department. Human CD34 +cells were isolated by the method of magnetic-activated cell sorting microbeads. Bone marrow mononuclear cells were isolated by Ficoll-Paque centrifugation,and the bone marrow mesenchymal stem cells(BMMSC) were cultured with L-DMEM. Human CD34 +cells and BMMSC were divided into 4 groups,and treated with PGE2(as positive control),DMSO(as negative control),EP2A and EP2A+prostaglandin E2receptor 2 antagonist (EP2AA),respectively. After exposed to the reagents,human CD34 +cell viability was measured by CCK-8 assay,the number of colonies was evaluated by colony-formation assay,the cell cycle distribution was analyzed by flow cytometry,and the protein expression of survivin,β-catenin and CXC chemokine receptor 4 (CXCR4) was detrmined by Western blot. Moreover, the concentration of stromal cell-derived factor-1α (SDF-1α) in the BMMSC was detected by ELISA. RESULTS:The cell viability and the colony number of human CD34 +cells in EP2A group were not higher than those in negative control group. Furthermore,the proportion of human CD34 +cells treated with EP2A in G2/M phase was not elevated compared with negative control group. The protein expression of survivin and β-cate-nin did not up-regulated in human CD34 +cells exposed to EP2A,but the protein expression of CXCR4 in human CD34 +cells and the concentration of SDF-1α in BMMSC were elevated. CONCLUSION:EP2A promotes human CD34 +cell homing in vitro but not proliferation.
ABSTRACT
Objective:To investigate the effects and its mechanism of lipopolysaccharide on the proliferation,differentiation and apoptosis of human dental pulp stem cells by Notch signaling pathway. Methods:Human dental pulp stem cells( hDPSCs) was isolated from dental pulp tissue;cell proliferation was detected after 0,0. 1,1,10 μg/ml treated cells for 1,3,5,7 days by CCK8 test;related mRNA expression ALP,DSPP,DMP1 gene was detected after 1 μg/ml lipopolysaccharide treated hDPSCs for 0,3,7,14,21 day by RT-PCR;cell apoptosis was detected after 1 μg/ml lipopolysaccharide treated hDPSCs for 0,7,14,21 day by flow cytometry. Cleaved caspase3,Notch1,Hes1 protein expression was detected by Western blot. Results:Cell proliferation after different concentrations lipopo-lysaccharide stimulated hDPSCs for 1, 3, 5 days had no significant difference, significantly lower at 7 day than 0 μg/ml lipopolysaccharide group(P<0. 01). ALP,DSPP,DMP1 mRNA expression lipopolysaccharide treated hDPSCs at 3 day compared with the control group had no statistical significance(P>0. 05),significantly higher at 7,14,21 day than control group(P<0. 01);cells apoptosis rate and Cleaved caspase3,Notch1,Hes1 protein expression lipopolysaccharide treatment hDPSCs at 7,14 and 21day was sig-nificantly higher than the control group(P<0. 01);ALP,DSPP,DMP1 mRNA expression and apoptosis rate and Cleaved caspase3, Notch1,Hes1 protein expression at 21 day was a downward trend. Conclusion:Lipopolysaccharide can decrease the proliferation of hDPSCs and promote its mineralization and apoptosis,which may be related to the activation of Notch signaling pathway.
ABSTRACT
Objective:To investigate the effects and its mechanism of lipopolysaccharide on the proliferation,differentiation and apoptosis of human dental pulp stem cells by Notch signaling pathway. Methods:Human dental pulp stem cells( hDPSCs) was isolated from dental pulp tissue;cell proliferation was detected after 0,0. 1,1,10 μg/ml treated cells for 1,3,5,7 days by CCK8 test;related mRNA expression ALP,DSPP,DMP1 gene was detected after 1 μg/ml lipopolysaccharide treated hDPSCs for 0,3,7,14,21 day by RT-PCR;cell apoptosis was detected after 1 μg/ml lipopolysaccharide treated hDPSCs for 0,7,14,21 day by flow cytometry. Cleaved caspase3,Notch1,Hes1 protein expression was detected by Western blot. Results:Cell proliferation after different concentrations lipopo-lysaccharide stimulated hDPSCs for 1, 3, 5 days had no significant difference, significantly lower at 7 day than 0 μg/ml lipopolysaccharide group(P<0. 01). ALP,DSPP,DMP1 mRNA expression lipopolysaccharide treated hDPSCs at 3 day compared with the control group had no statistical significance(P>0. 05),significantly higher at 7,14,21 day than control group(P<0. 01);cells apoptosis rate and Cleaved caspase3,Notch1,Hes1 protein expression lipopolysaccharide treatment hDPSCs at 7,14 and 21day was sig-nificantly higher than the control group(P<0. 01);ALP,DSPP,DMP1 mRNA expression and apoptosis rate and Cleaved caspase3, Notch1,Hes1 protein expression at 21 day was a downward trend. Conclusion:Lipopolysaccharide can decrease the proliferation of hDPSCs and promote its mineralization and apoptosis,which may be related to the activation of Notch signaling pathway.