Korean J Pediatr.  2008 Feb;51(2):170-180. 10.3345/kjp.2008.51.2.170.

Neuroprotective effects of geneticin (G418) via apoptosis in perinatal hypoxic-ischemic brain injury

• Ju M
• Lee HJ
• Lee SJ
• Seo ES
• Park HJ
• Lee KY
• Lee GH
• Choi EJ
• Kim JK
• Lee JW
• Chung HL
• Kim WT

• KMID: 2280226
• DOI: http://doi.org/10.3345/kjp.2008.51.2.170

Abstract

PURPOSE: Some antibiotics were known to exert neuroprotective effects in the animal model of hypoxic-ischemic (H-I) brain injury, but the mechanism is still unclear. A recent study reported that geneticin (G418), an aminoglycoside antibiotic, increased survival of human breast cancer cells by suppressing apoptosis. We investigated the neuroprotective effects of systemically administrated geneticin via anti-apoptosis following the H-I brain injury
METHODS
Seven-day-old Sprague-Dawley rat pups were subjected to unilateral (left) common carotid artery occlusion followed by 2.5 hours of hypoxic exposure and the cortical cell culture of rat brain was done under a hypoxic incubator. Apoptosis was measured in the injured hemispheres 7 days after H-I insult and in the injured cells from hypoxic chamber using morphologic analysis by Terminal dUTP Nick-end Labeling(TUNEL) assay and immunohistochemistry for caspase-3, and cytologic analysis by western blot and real time PCR for bax, bcl-2, and caspase-3.
RESULTS
The gross appearance and hematoxylin and eosin stain revealed increased brain volume in the geneticin-treated animal model of perinatal H-I brain injury. The TUNEL assay revealed decreased apoptotic cells after administration of geneticin in the cell culture model of anoxia. Immunohistochemistry showed decreased caspase-3 expression in geneticin-treated cortical cell culture. Western blot and real-time PCR showed decreased caspase-3 expression and decreased ratio of Bax/Bcl-2 expression in geneticin-treated animal model.
CONCLUSION
Geneticin appears to exert a neuroprotective effect against perinatal H-I brain injury at least via anti-apoptosis. However, more experiments are needed in order to demonstrate the usefulness of geneticin as a preventive and rescue treatment for H-I brain injuries of neonatal brain.

Keyword

Hypoxic-ischemic brain injury; Anti-apoptosis; Perinatal; Geneticin; G418

MeSH Terms

Animals
Anoxia
Anti-Bacterial Agents
Apoptosis
Blotting, Western
Brain
Brain Injuries
Breast Neoplasms
Carotid Artery, Common
Caspase 3
Cell Culture Techniques
Eosine Yellowish-(YS)
Gentamicins
Hematoxylin
Humans
Immunohistochemistry
In Situ Nick-End Labeling
Incubators
Models, Animal
Neuroprotective Agents
Rats
Real-Time Polymerase Chain Reaction
Anti-Bacterial Agents
Caspase 3
Eosine Yellowish-(YS)
Gentamicins
Hematoxylin
Neuroprotective Agents