Blue light irradiation can induce skin pigmentation by causing oxidative stress in melanocytes. Researches have demonstrated that Nicotinamide Nucleotide Transhydrogenase (NNT) located in mitochondrial inner membrane regulates pigmentation, which is also related to redox homeostasis. In order to investigate the potential relationship between these two pathways, this study utilized mouse melanoma cell B16-F10 and human melanoma cell MNT1 to establish models of two pigmentation models of Nnt (NNT) knockdown and blue light irradiation. The effects of blue light irradiation on Nnt (NNT) expression and distribution were assessed, and initially explored the effects of both induction conditions on mitochondrial function and morphology. Moreover, the colocalization of mitochondria and melanosomes under blue light induced model was observed. The results indicated that both blue light irradiation and Nnt (NNT) knockdown led to an increase of melanin in melanocytes. And blue light irradiation down-regulated the expression of Nnt (NNT) and the protein was localized to the nucleus. Assessment of mitochondrial membrane potential revealed that both blue light exposure and Nnt (NNT) knockdown could reduce mitochondrial membrane potential. Furthermore, analysis of mitochondrial dynamics-related proteins demonstrated that blue light exposure and Nnt knockdown slightly elevated the expression of certain mitochondrial fusion-related proteins (MFN1, MFN2, OPA1) and down-regulated the expression of the mitochondrial fission-related protein DRP1 in B16-F10 cells. Co-localization staining of mitochondria and melanosomes showed an extensive distribution, with increased co-localization in the perinuclear region after blue light irradiation. This study provides initial insights into the potential relationship between mitochondria and melanin synthesis, offering a novel perspective for exploring the regulatory mechanisms of pigmentation.