Fig 1.
Bilirubin and its photoderivatives.
(A) Z,Z-Bilirubin IXα. (B) Z,E-Bilirubin IXα. (C) E,Z-Bilirubin IXα. (D) Z-Lumirubin IXα.
Fig 2.
Compounds produced by bilirubin phototherapy.
(A) TLC plate after first chromatography. (B) Most important compounds 1 and 7 were separated by re-chromatography from the 1st (upper panel), and 7th zone (lower panel). UCB, unconjugated bilirubin.
Fig 3.
HPLC chromatograms of isolated bilirubin PI.
(A) HPLC chromatogram of band 1 from TLC—mixture of ZE/EZ-bilirubins; peak 1 = EZ-bilirubin, peak 2 ZE-bilirubin. (B) HPLC chromatogram of band 7 from TLC—Z-lumirubin.
Table 1.
List of genes used for gene expression analyses.
Fig 4.
Effect of bilirubin PI on Bf concentrations.
Table 2.
The binding constant for the ligand-HSA complexes.
Fig 5.
(Left panel) CD and UV/Vis absorption spectra of unbound (broken line), lumirubin (blue), bilirubin (red), and their complexes with HSA (full line). Pigment/HSA molar ratio = 1/1; c (pigment) = 1.5 x 10−5 mol/L. Bilirubin was dissolved in 0.1 mol/L NaOH and mixed with the HSA solution in PBS (pH 7.4) at the molar ratio [pigment]/[HSA] = 1/1, the concentration of the pigment was 1.5 x 10−5 mol/L. (Right panel) Effect of lumirubin on HSA binding with different marker ligands: bilirubin (A), hemin (B), and with both bilirubin and hemin (C). CD spectra of HSA-marker ligand are shown in green, lumirubin-HSA complex and lumirubin bound to the complex HSA-marker ligand are shown as blue and black full lines, respectively. Black arrows show the changes in signals after formation of lumirubin-(marker ligand-HSA) complexes. Lumirubin/HSA/marker molar ratio = 1/1/1.
Fig 6.
The effect of bilirubin and bilirubin PI on the cell viability in our in vitro model.
(A) 24 hr exposure; n = 7. (B) 48 hr exposure; n = 4. * p < 0.05 vs. control; # p < 0.05 vs. bilirubin. Cell viability tested by CellTiter-Glo test; exactly the same results were obtained with MTT test. UCB, unconjugated bilirubin.
Table 3.
The effect of bilirubin and bilirubin PI on expression of selected genes.