Effect of steroids on apoptosis and neuropeptide receptor gene expression of human bone marrow mesenchymal stem cells

Abstract

Objective To observe the effect of steroids on the expression of calcitonin gene-related peptide receptor (CGRPR),substance P receptor (SPR),perxisome proliferators activated receptor-γ (PPARγ),Runx2,B lymphocytes/leukemia-2 (bcl-2) and Caspase-3 in human bone marrow mesenchymal stem cells (hBMSCs).Methods hBMSCs were isolated and purified by density gradient centrifugation combined with plastic-adhering culture,and identified by flow cytometry.The hBMSCs of the 3rd generation were randomly divided into two groups.The control group received no treatment,while cells in the experimental group were treated with 1 × 10-7 mol/L dexamethasone.The mRNA was isolated and the expression level of bcl-2,Caspase-3,CGRPR,SPR,Runx2 and PPARγ was detected by using RT-qPCR on the 7th day after treatment.Results After isolation and culture,the hBMSCs displayed a distinct phenotype-the spindle-shaped,and uniform cells grew into clusters with swirl or radiation pattern.The expression levels of Caspase-3 and PPARγ in the experimental group were significantly higher than those in the control group,with the 2-△△Ct being 5.99 ±2.00 and 6.93 ±0.70 respectively (P ＜0.01).The expression levels of bcl-2,CGRPR,SPR and Runx2 were lower in the experimental group than those in the control group,with the 2-△△Ct being 0.20 ±0.51,0.11 ±0.02,0.28 ±0.08 and 0.13 ±0.21 respectively (P ＜0.01).Conclusion The steroid can up-regulated the expression of apoptosis gene and adipogenic gene in hBMSCs,and down-regulated the expression of anti-apoptosis gene and neuropeptide factors,which can promote the apoptosis and adipogeneic differetiantion of the cells and inhibit their osteogenic differentiantion. Key words: s] Human bone marrow mesenchymal stem cells; Steroid; Neuropeptide; Apoptosis