RESUMEN
BACKGROUND: Oxidative stress plays an important role in the skin aging process. Rapamycin has been shown to have anti-aging effects, but its role in oxidative senescence of skin cells remains unclear. The aim of this study was to explore the effect of rapamycin on oxidative stress-induced skin cell senescence and to illustrate the mechanism. METHODS: Primary human skin fibroblasts (HSFs) were extracted and a model of H2O2-induced oxidative senescence was constructed, and the effects of rapamycin on their value-added and migratory capacities were detected by CCK-8 and scratch assays. SA-ß-gal was utilized to detect senescence, oxidatively closely related factors were also assessed. Gene and protein expressions of senescence, oxidative, and autophagy were detected by western blotting and quantitative-PCR. The data were analyzed by one-way analysis of variance. RESULTS: Rapamycin (0.1 nmol/L for 48 h) promoted the proliferative and migration of H2O2-treated HSFs (p < 0.05), decreased senescent phenotypes SA-ß-gal staining and the expression of P53, and MMP-1 proteins, and increased the expression level of COL1A-1 (p < 0.001). Rapamycin also enhanced the activities of SOD and HO-1, and effectively removed intracellular ROS, MDA levels (p < 0.05), in addition, autophagy-related proteins and genes were significantly elevated after rapamycin pretreatment (p < 0.001). Rapamycin upregulated the autophagy pathway to exert its protective effects. CONCLUSION: Our findings indicate that rapamycin shields HSFs from H2O2-induced oxidative damage, the mechanism is related to the reduction of intracellular peroxidation and upregulation of autophagy pathway. Therefore, rapamycin has the potential to be useful in the investigation and prevention of signs of aging and oxidative stress.
Asunto(s)
Autofagia , Senescencia Celular , Fibroblastos , Peróxido de Hidrógeno , Estrés Oxidativo , Sirolimus , Piel , Humanos , Sirolimus/farmacología , Peróxido de Hidrógeno/farmacología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Senescencia Celular/efectos de los fármacos , Piel/efectos de los fármacos , Piel/metabolismo , Autofagia/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Especies Reactivas de Oxígeno/metabolismoRESUMEN
BACKGROUND/PURPOSE: Chlorhexidine (CHX) is a type of chemical antiseptic that is widely used in dental practice. Stem cells from human exfoliated deciduous teeth (SHED) are multipotent cells. However, there is little knowledge about the effects of chlorhexidine on SHED cells. The purpose of this study is to investigate the effects of CHX on SHED. METHODS: SHED cells were treated with 0.1%, 0.01%, 0.001%, and 0.0001% CHX for 10 seconds to test the effects of different concentrations of CHX on SHED cells. The cells were also treated with 0.01% CHX for 10 seconds, 1 minute, and 5 minutes to test the time effects of CHX on SHED cells. Cell proliferation was investigated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and an autonomously replicating sequence (ARS) assay was used for the evaluation of the mineralization potential. RESULTS: This study demonstrated that different concentrations of CHX had cytotoxic effects on SHED cells in a dose- and time-dependent manner. The proliferation of SHED cells was inhibited by approximately 50% by the use of 0.01% CHX. It was also found that the cell proliferation and mineralization potential of SHED cells were inhibited to some degree by different concentrations of CHX. CONCLUSION: Different concentrations of CHX can inhibit SHED cell proliferation in a dose- and time-dependent manner. In addition, the mineralization potential of SHED cells is inhibited to some degree by different concentrations of CHX.