The Effects Of Hydrogen Sulphide On Adipocyte Viability In Human Adipocyte and Adipocyte Derived Mesenchymal Stem Cells Culture Under Ischemic Conditions

Hydrogen Sulfide (H₂S) is accepted as a gasotransmitter, which has a gaseous structure like the other two gasotransmitters; carbon monoxide and nitric oxide. It is known that H₂S increases the resistance to ischemia in some tissues and decreases apoptosis, however the exact mechanism is still unclear. Morover there is no research about its effects on adipose tissue in the literature.

The aim of this study was to evaluate the in vitro effects of H₂S on Adipocyte survival under ischemic conditions and to explain the possible mechanisms of its apoptotic process in details.

For this purpose mesenchymal stem cell (MSCs) culture was prepared from subcutaneous adipose tissue sample of a human donor. Adipose derived MSCs were differentiated into adipogenic direction and mature adipocyte culture was obtained. Adipose derived MSCs culture and mature adipocyte culture were divided into six groups separately. Sodium Hydrogen Sulfide (NaHS) was used as a H₂S donor. After applying NaHS to all groups with a concentration of 0, 0.1, 1, 10, 100 and 1000μM, cell cultures were incubated in special gas incubator with %1 oxygen at 37°C for 24 hours. Following the ischemia period, cell culture groups were evaluated by the rates of proliferation/cytotoxicity within MTT test, apoptosis and necrosis rates with flow cytometry, apoptotic (bax, caspase3) and anti-apoptotic genes (bcl-2) expression levels within reverse transcriptase polymerase chain reaction.

Statistically significant increase in proliferation rates were found in MSCs groups which was applied low dose (0,1 and 1 μM) NaHS (p<0,05). For every different dose applications, statistically significant decrease was found in 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.

As a conclusion, H₂S has protective effect on both MSCs and mature adipocytes and this effect is mediated by elevation of anti-apoptotic gene expression. Furthermore, this study suggests that, administration of H₂S to fat graft before fat graft injection may be therapeutic benefit in the fat graft viability.