Effect of hypoxia and serum deprivation on endogenous hydrogen sulfide production in rat bone marrow mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Ischemia microenvironment contributes mostly to the low survival rate of rat bone marrow mesenchymal stem cells after transplantation. Hydrogen sulfide (H2S) can protect various cells and tissue models against apoptosis and injury. OBJECTIVE:To detect the cellapoptosis and viability, content of H2S in supernatant, and the expression of H2S synthetase after different time of hypoxia and serum deprivation cultivation of rat bone marrow mesenchymal stem cells. METHODS:The passage 3 rat bone marrow mesenchymal stem cells were divided into five different cultivation time groups0-, 3-, 6-, 12-and 24-hour groups. After enough hypoxia and serum deprivation cultivated time, the cellapoptosis was detected by SubG1, the cellviability was determined by cellcounting kit-8, the content of H 2S in supernatant was measured by N,N-dimethyl-p-phenylenediamin and the expression of H2S synthetase by RT-PCR and western blot. RESULTS AND CONCLUSION:Compared to the normal cultivation group, after different hypoxia and serum deprivation cultivated time, the cellapoptosis increased and cellviability decreased significantly. The longer hypoxia and serum deprivation cultivated time caused the more cellapoptosis and the lower cellviability. The contents of H2S and its synthetase were also suppressed by hypoxia and serum deprivation cultivation. The difference was statistical y significant. These findings suggest that hypoxia and serum deprivation cultivation can inhibit the generation of H 2 expression of its synthetase.