*3.6. In Vitro Reduction of ROS*

The DGA regimen effectively hinders excessive ROS generation and possesses a similar or superior equimolar antioxidant effect to glutathione, and vitamins C and E in hepatocytes (Supplementary Materials). Similar ROS scavenging results were obtained from rat primary optic astrocytes, both in naturally induced and hydrogen peroxide induced ROS models (Supplementary Materials).

#### **4. Discussion**

Blood bilirubin concentration increased significantly 12 h after the last DGA on Day21 in the DGA group but not in the placebo group. However, the increase in blood Fe and the mRNA expression of the HO-1 gene in WBCs were not statistically significant. As expected, the increase in blood bilirubin during the 21-day DGA regimen was associated with reduced blood TGs and systemic inflammation. In the acute 45 min test, HO-1 enzyme activity seemed to be activated, based on responses in blood bilirubin and Fe levels. In the 4-day comparisons intra-individual responses of blood bilirubin and albumin correlated very strongly in the LC subgroup, indicating an activation of hepatic bilirubin intake. Furthermore, hepatic inflow of Fe was possibly activated in the LC subgroup during the 4-day DGA regimen. Finally, the changes in blood TGs and GlycA between the days 4 and 21 correlated inversely with bilirubin (and Fe).
