The Effects of Hydrogen Sulfide on Adipocyte Viability in Human Adipocyte and Adipocyte-Derived Mesenchymal Stem Cell Cultures Under Ischemic Conditions.

Abstract

This study evaluated the in vitro effects of hydrogen sulfide on adipocyte survival under ischemic conditions and explored possible mechanisms of its apoptotic process. The mesenchymal stem cell culture was prepared from a human subcutaneous adipose tissue sample. Adipose-derived mesenchymal stem cells were differentiated into the adipogenic direction, and a mature adipocyte culture was obtained. The adipose-derived mesenchymal stem cell and mature adipocyte cultures were both divided into 6 groups. Sodium hydrogen sulfide was used as a hydrogen sulfide donor. After treating the groups with sodium hydrogen sulfide (0, 0.1, 1, 10, 100, and 1000 μM), the cell cultures were incubated in 1% oxygen at 37°C for 24 hours. After the ischemia period, the cell culture groups were evaluated with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test for the proliferation/cytotoxicity rates, flow cytometry for apoptosis and necrosis rates, and reverse transcriptase polymerase chain reaction for apoptotic (Bax, Caspase-3) and antiapoptotic (Bcl-2) gene expression levels. Statistically significant increases in proliferation rates were found in mesenchymal stem cell groups treated with low dose (0, 1, and 1 μM) sodium hydrogen sulfide (P<0.05). For each dose, a statistically significant decrease was found in late apoptosis levels on the mature adipocyte cultures (P<0.05). In both cell culture groups, Bcl-2 gene expression was increased and Caspase-3 gene expression was decreased. Under ischemic conditions, hydrogen sulfide has a protective effect on mesenchymal stem cells and mature adipocytes, and this effect is mediated by the elevation of antiapoptotic gene expression.