J Korean Neurol Assoc.  2003 Aug;21(4):392-400.

Cytoprotective Effect of Epigallocatechin gallate (EGCG) in Oxidative-stressed PC12 Cells Following H2O2 Exposure-Effect of EGCG on Phosphoinositide 3-kinase/Akt and Glycogen Synthase Kinase-3 Pathway

• Koh SH
• Kim SH
• Kwon H
• Song CW
• Kim JH
• Kim J
• Chung B
• Kim MH
• Gong G
• Yu HJ
• Jung HK

Abstract

BACKGROUND
Neurodegenerative diseases (ND) are associated with oxidative stress, and antioxidants including epigallocatechin gallate (EGCG) have been tried as potential therapeutic regimens of an experimental model of ND. We performed this study to determine the neuroprotective role of EGCG on up stream and down stream signals in oxidative-stress-injured PC12 cells by exposing them to H2O2. METHODS: Following 100 microM H2O2 exposure, the viability of PC12 cells (not pretreated vs EGCG or z-VAD-fmk pretreated) was evaluated by using a MTT assay. Immunoreactivity (IR) of cytochrome c, caspase-3, poly (ADP-ribose) polymerase (PARP), PI3K/Akt and GSK-3 was examined by using a Western blot. RESULTS: EGCG or z-VAD-fmk pretreated PC12 cells showed increased viability. Dose-dependent inhibition of caspase-3 activation and PARP cleavage was demonstrated by the pretreatment of both agents. However, the inhibition of cytochrome c release was only detected in EGCG pretreated cells. On the pathway through PI3K/Akt and GSK-3, however, the result of a western blot in EGCG pretreated cells showed decreased IR of Akt and GSK-3 and increased IR of p85a PI3K, phosphorylated Akt and GSK-3, and contrasted with that in z-VAD-fmk pretreated cells showing no changes. CONCLUSIONS: These data show that EGCG affects apoptotic pathways through upstream signals including PI3K/Akt and GSK-3 pathways as well as downstream signals including cytochrome c and caspase-3 pathways. Therefore, these results suggest that EGCG mediated activation of PI3K/Akt and inhibition GSK-3 could be a new protective mechanism on the pathogenesis of ND.

Keyword

Apoptosis; Neurodegenerative disease; Antioxidants; Glycogen synthase kinase 3; PI3K; Akt

MeSH Terms

Animals
Antioxidants
Apoptosis
Blotting, Western
Caspase 3
Cytochromes c
Glycogen Synthase Kinase 3
Glycogen Synthase*
Glycogen*
Models, Theoretical
Neurodegenerative Diseases
Oxidative Stress
PC12 Cells*
Rivers
Antioxidants
Caspase 3
Cytochromes c
Glycogen
Glycogen Synthase
Glycogen Synthase Kinase 3