The Protective Effect of Crocin Against Induced Oxidative Stress on Adipose-Derived Stem Cells (ADSC)

Background and Aim: Adipose-derived stem cell (ADSC) is broadlyused in cell transplantation. It is easily obtained by a minimally invasiveprocedure, genetically stable in long-term culture and capable ofevading the host’s immune system. As survival and longevity of fat graftsalong with ADSCs in various tissues remain poor post-engraftment, usingpractical methods such as natural anti-oxidants to enhance their survivalhas a crucial role to improve the efficiency of fat grafting. Therefore, weconducted this study to evaluate the protective effect of crocin againstoxidative stress induced by H2O2 on ADSCs in vitro.Methods: ADSCs were treated with different concentrations of crocinand H2O2, separately. Then the MTT assay was used to determine the IC50of crocin and H2O2. LDH assay was carried out to evaluate the H2O2-induced cytotoxicity in cultured ADSCs. After treatment of these cellswith specific concentrations of crocin and H2O2, intracellular level ofROS was measured based on the oxidation of non-fluorescent probe2′,7′-dichlorodihydrofluorescein diacetate (H2DCF-diacetate) to yieldthe highly fluorescent DCF. Also, cell survival was measured by thetrypan blue assay and DAPI (4′,6-diamidino-2-phenylindole) staining.Then the signaling pathway of Nrf2/ HO1 in ADSCs treated with crocinand H2O2was studied by real-time PCR.Results: We investigated ADSCs survival with and without crocinpretreatment under oxidative conditions. Crocin prevented cell death ina dose-dependent manner with the concentration of 10 μM exhibited a maximal protective effect against H2O2-induced cytotoxicity (P < 0.05vs. H2O2 group). Additionally, cell viability did not change by incubationwith crocin (1–10 μM) in H2O2-free condition. H2O2 markedly inducedthe release of LDH, while crocin pretreatment dose-dependentlysuppressed LDH release in ADSCs. According to trypan blue assay andDAPI staining, crocin increased the cell survival dose-dependently. It hasbeen shown that crocin is involved in Nrf2 signaling pathway in ADSCs.This study showed crocin protects the ADSCs against oxidative stressinduced by H2O2.Conclusion: In this study, the pivotal role of crocin as a powerful andnatural anti-oxidant against oxidative stress on Adipose-Derived StemCells (ADSCs) has been shown. Our findings provide a novel method forincreasing the survival of ADSCs used in fat-grafting in tissues.

کلیدواژه ها:

Adipose-derived stem cell (ADSC) ، Oxidative stress ، Crocin