1. CHLOROGENIC ACID AND COFFEE PREVENT HYPOXIA- INDUCED RETINAL DEGENERATION Holim Jang, Hong Ryul Ahn, Hyoung Jo, Kyung-A Kim, Eun Ha Lee, Ki Won Lee, Sang Hoon Jung, and Chang Y. Lee

2. The Retina  Photographic film of the eye  Retinal ganglion (RGC-5) cells  Axons form optic nerve

3. Hypoxia  Metabolically active  Oxidative stress  Reduction in oxygen supply  Risk factor for sight-threatening disorders

4. Nitric Oxide (NO)  Hypoxia  NO binds to Complex IV  O 2 - formed  Formation of peroxynitrite (oxidizing agent)  NO + O 2 -  ONO 2 -

5. Chlorogenic Acid (CGA)  Antioxidant phytochemical  Major source is coffee  High blood absorption rate (33%)  Green coffee

6. Question  Do CGA and coffee protect against retinal degeneration?  In vitro & in vivo

7. Method: In Vivo  Male mice 30-35 kg  Optic nerve crush  Left: crush  Right: control  CGA & coffee extract (oral)

8. Method: In Vivo  RGC labeling: superior colliculi exposed  Fluorogold  Eyeballs enucleated after 5 days  Retinal flat mounts

9. Method: Analysis  Hematoxilin & Eosin staining  Nuclei = blue  Cytoplasm & proteins = pink  TUNEL staining  Apoptosis = brown

10. Results: CGA

14. Results: Coffee Extract

16. Results: Drawing Conclusions  CGA’s NO scavenging activity  Optic nerve damage  inflammatory response  NO release  Supported by in vitro

17. Discussion  Retinal degeneration from hypoxia  CGA = NO scavenger  Limitations  Mice  CGA concentration