Blockade of Cyclophilin D Attenuates Oxidative Stress-Induced Cell Death in Human Dental Pulp Cells
H2O2-induced cell death and mitochondrial dysfunction in the HDPCs. (a) Cell viability was determined by MTT reduction in the HDPCs with or without the presence of H2O2. (b, c) Flow cytometric quantification of cell death. HDPCs were treated for 24 h with culture medium or H2O2 (150 μM, 250 μM). (d, e) TUNEL staining and assay. (f) Representative immunoreactive bands with different densities for Bcl-2 and Bax in the HDPCs in the presence of H2O2. Quantification of immunoreactive bands for Bax (g) and Bcl-2 (h) relative to β-actin. Representative images showing MitoSOX staining (i) and quantification (j) in the indicated groups. Representative images with TMRM staining (k) and quantification (l) in the indicated groups. Representative images showing Fluo-4-AM staining (m) and quantification (n) in the indicated groups. ATP (o) in the indicated groups. (p) Densitometry of immunoreactive bands for CypD in the HDPCs in the presence of H2O2. (q) Quantification of immunoreactive bands for CypD relative to β-actin. HDPCs were treated for 24 h with H2O2 (250 μM) (+) or culture medium (-). Data represent the mean of three independent experiments.
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