title: Effects of cerebrolysin on motor-neuron-like NSC-34 cells.
abstract: Although the peripheral nervous system is capable of regeneration, this capability is limited. As a potential means of augmenting nerve regeneration, the effects of cerebrolysin (CL)--a proteolytic peptide fraction--were tested in vitro on the motor-neuron-like NSC-34 cell line and organotypic spinal cord cultures. Therefore, NSC-34 cells were subjected to mechanical stress by changing media and metabolic stress by oxygen glucose deprivation. Afterwards, cell survival/proliferation using MTT and BrdU-labeling (FACS) and neurite sprouting using ImageJ analysis were evaluated. Calpain-1, Src and α-spectrin protein expression were analyzed by Western blot. In organotypic cultures, the effect of CL on motor neuron survival and neurite sprouting was tested by immunohistochemistry. CL had a temporary anti-proliferative but initially neuroprotective effect on OGD-stressed NSC-34 cells. High-dosed or repeatedly applied CL was deleterious for cell survival. CL amplified neurite reconstruction to limited extent, affected calpain-1 protein expression and influenced calpain-mediated spectrin cleavage as a function of Src expression. In organotypic spinal cord slice cultures, CL was not able to support motor neuron survival/neurite sprouting. Moreover, it hampered astroglia and microglia activities. The data suggest that CL may have only isolated positive effects on injured spinal motor neurons. High-dosed or accumulated CL seemed to have adverse effects in treatment of spinal cord injury. Further experiments are required to optimize the conditions for a safe clinical administration of CL in spinal cord injuries.
authors.affiliation: Institute of Biochemistry and Cell Biology, Otto-von-Guericke University Magdeburg, Leipziger Str. 44, D-39120 Magdeburg, Germany. Electronic address: Gerburg.keilhoff@med.ovgu.de.
authors.forename: Gerburg
authors.initials: G
authors.lastname: Keilhoff
authors.affiliation: Institute of Biochemistry and Cell Biology, Otto-von-Guericke University Magdeburg, Leipziger Str. 44, D-39120 Magdeburg, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Lucas
authors.affiliation: Institute of Biochemistry and Cell Biology, Otto-von-Guericke University Magdeburg, Leipziger Str. 44, D-39120 Magdeburg, Germany.
authors.forename: Josephine
authors.initials: J
authors.lastname: Pinkernelle
authors.affiliation: Institute of Biochemistry and Cell Biology, Otto-von-Guericke University Magdeburg, Leipziger Str. 44, D-39120 Magdeburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Steiner
authors.affiliation: Department of Plastic, Reconstructive and Aesthetic Surgery, Hand Surgery, Klinikum Bielefeld, Teutoburger Str. 50, D-33604 Bielefeld, Germany.
authors.forename: Hisham
authors.initials: H
authors.lastname: Fansa
mesh_terms: D000596:Amino Acids / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D002154:Calpain / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D007124:Immunoenzyme Techniques; D051379:Mice; D009046:Motor Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D009416:Nerve Regeneration / Q000187:drug effects; D016501:Neurites / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009924:Organ Culture Techniques; D013049:Spectrin / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D000596:Amino Acids; D018696:Neuroprotective Agents; D013049:Spectrin; C006952:cerebrolysin; D019061:src-Family Kinases; D002154:Calpain; C410233:Capns1 protein, mouse
keywords: BrdU; Calpain; Cerebrolysin; NSC-34 cells; Organotypic spinal cord culture; Spectrin; Src