PMC

The effect of adrenochrome and BNAH on ROS production in HT-22 cells. Cells were incubated with adrenochrome (50–200 µM) and/or BNAH (100 µM) for 15 min. Final concentration of DMSO was 0.2%. Data are presented as median (min–max). All experiments were performed in 12 replicates. Kruskal–Wallis test with Dunn’s post-hoc: * p < 0.05, ** p < 0.01 vs. Adrenochrome (0 μM) + BNAH group in ROS measurement with CellROX Green Reagent; # p < 0.05, ## p = 0.003, ### p < 0.001 vs. Adrenochrome (0 μM) + BNAH group in ROS measurement with CM-H₂DCFDA. For details see .

(a) Fontana Masson stain of corneal biopsy demonstrating pigmented granules that stain positively, indicating the presence of adrenochrome ( × 20). (b) Hematoxylin and eosin stain corneal biopsy demonstrating pigmented, acellular intrastromal material ( × 20). Epithelium and Bowman's membrane are absent, consistent with clinical defect. Gram stain demonstrates diffuse intralamellar Gram-positive cocci (insert, arrow).

Comparisons of QR2 and QR1 activities in the reduction of adrenochrome. A, reduction of adrenochrome using SUB10R as the electron donor. Progression of the reaction was determined by monitoring the consumption of electron donors, which corresponds to UV light absorbance decrease at wavelengths around 340 nm. B, NMN was used as the electron donor. C, when NADH was used as the electron donor, neither QR1 nor QR2 exhibited any activity. D, saturation curve of the reduction of adrenochrome by QR2 using SUB10R as the electron donor.

The influence of pre-incubation with NQO2 inhibitors on nuclear DNA damage of HT-22 cells exposed to adrenochrome and BNAH for 120 min. Cells were pre-incubated with M-11 (a) or S29434 (b) (100 µM) for 30 min, and then were exposed to adrenochrome (125 or 200 µM) and BNAH (100 µM) for 120 min. Adrenochrome and BNAH were dissolved in DMSO. Final concentration of DMSO was 2%. All experiments were performed in 6 replicates. Data are presented as mean ± S.D. The percentage of DNA in the comet tail (blank bars) of cells exposed to adrenochrome and BNAH was significantly higher than DMSO (dotted line) with p < 0.001 (one-way ANOVA with Tukey’s post-hoc). Data on the percentage of atypical comets (colored bars) were evaluated using Kruskal-Wallis test with Dunn’s post-hoc: + p < 0.01 vs. DMSO. To evaluate the differences between groups exposed to adrenochrome and BNAH, two-way ANOVA with Tukey’s post-hoc was used: * p < 0.001 vs. pre-incubation with M-11 or S29434. For details see .