Neuroinflammation and Neurodegeneration in Adult Rat Brain from Binge Ethanol Exposure: Abrogation by Docosahexaenoic Acid
Figure 4
Effects of binge ethanol treatment and DHA supplementation on neurodegeneration and oxidative (3NT-) protein footprints in adult-age HEC slice cultures.
Binge ethanol treatment of adult-age rat HEC slice cultures for 4 days as described in Methods significantly increases degenerating neurons (PI staining) and the levels of oxidative stress footprints (3NT-proteins), and supplementation of cultures with DHA prevents the increases. Fig. 4A: (top) Representative PI-stained images showing increased neurodegeneration after 3 days of binge ethanol treatment (100 mM), with DHA supplementation (25 µM) suppressing PI staining (neurodegeneration). (Bottom) Quantitation of PI labeling shows significant neurodegeneration due to 4 days of binge ethanol exposure (E), and neuroprotection against E by DHA at 25 and 50 µM. **p<0.01 vs. C. #p<0.05 vs. E. Fig. 4B. Quantitation of PI labeling reveals significantly increased neurodegeneration over control due to binge E as early as 2 days of binge ethanol treatment, with prevention of the neurodegeneration throughout the 4 days of treatment by DHA supplementation. *p<0.05 vs. control (C) or E+DHA. Fig. 4C (Top) Representative immunoblots of 3NT-proteins in HEC slice cultures following binge ethanol exposure (100 mM) for 4 days. (Bottom) Quantitation of immunoblots showing that binge ethanol exposure causes increased 3NT-proteins in the HEC slice cultures, and DHA supplementation (25 µM) prevents the increases. *p<0.05 vs. Control.