Estrogen treatment of spinal cord injury attenuates calpain activation and apoptosis.
Spinal cord injury (SCI) is a devastating neurologic injury, and currently, the only recommended pharmacotherapy is high-dose methylprednisolone, which has limited efficacy. Estrogen is a multi-active steroid with anti-oxidant and anti-apoptotic effects. Estrogen may modulate intracellular Ca2+ and prevent inflammation. For this study, male rats were divided into three groups. Sham-group animals received a laminectomy at T12. Injured rats received both laminectomy and 40 gram centimeter force SCI. Estrogen-group rats received 4 mg/kg 17beta-estradiol (estrogen) at 15 min and 24 hr post-injury, and vehicle-group rats received equal volumes of dimethyl sulfoxide. Animals were sacrificed at 48 hr post-injury, and 1-cm segments of the lesion, rostral penumbra, and caudal penumbra were excised. The degradation of 68 kD neurofilament protein (NFP) and estrogen receptors (ER) was examined by Western blot analysis. Protein levels of calpain and the activities of calpain and caspase-3 were also examined. Levels of cytochrome c were determined in both cytosolic and mitochondrial fractions. Cell death with DNA fragmentation was examined using the TUNEL assay. At the lesion, samples from both vehicle and estrogen treated animals showed increased levels of 68 kD NFP degradation, calpain content, calpain activity, cytochrome c release, and degradation of ERalpha and ERbeta, as compared to sham. In the caudal penumbra, estrogen treatment significantly attenuated 68 kD NFP degradation, calpain content, calpain activity, levels of cytosolic cytochrome c, and ERbeta degradation. At the lesion, vehicle-treated animals displayed more TUNEL+ cells, and estrogen treatment significantly attenuated this cell death marker. We conclude that estrogen may inhibit cell death in SCI through calpain inhibition.
Sribnick
Eric Anthony
EA
Department of Neurosciences, Division of Neurology, Medical University of South Carolina, Charleston, South Carolina 29425, USA. baniknl@musc.edu
Matzelle
Deborah Denise
DD
Ray
Swapan Kumar
SK
Banik
Naren Lal
NL
D000704:Analysis of Variance; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western / Q000379:methods; D001896:Boron Compounds / Q000378:metabolism; D002154:Calpain / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D004195:Disease Models, Animal; D004789:Enzyme Activation / Q000187:drug effects; D004967:Estrogens / Q000627:therapeutic use*; D005455:Fluorescent Antibody Technique / Q000379:methods; D020287:In Situ Nick-End Labeling / Q000379:methods; D007796:Laminectomy / Q000379:methods; D008297:Male; D016900:Neurofilament Proteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011960:Receptors, Estrogen / Q000378:metabolism; D013119:Spinal Cord Injuries / Q000188:drug therapy*; D013997:Time Factors
D001896:Boron Compounds; D004967:Estrogens; D016900:Neurofilament Proteins; D011960:Receptors, Estrogen; C066858:neurofilament protein NF 68; C045000:dicesium N-succinimidyl 3-(undecahydrododecaboranyldithio)propionate; D045304:Cytochromes c; D002154:Calpain; D053148:Caspase 3