Improvement in functional recovery with administration of Cerebrolysin after experimental closed head injury

Laboratory investigation

Yanlu Zhang M.S., M.D. 1 , Michael Chopp Ph.D. 2 , 3 , Yuling Meng Ph.D. 1 , Zheng Gang Zhang M.D., Ph.D. 2 , Edith Doppler Ph.D. 4 , Asim Mahmood M.D. 1 and Ye Xiong M.D., Ph.D. 1
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  • 1 Departments of Neurosurgery and
  • 2 Neurology, Henry Ford Hospital, Detroit;
  • 3 Department of Physics, Oakland University, Rochester, Michigan; and
  • 4 Clinical Research and Pharmacology, EVER Neuro Pharma GmbH, Unterach, Austria
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Object

Cerebrolysin is a unique peptide preparation that mimics the action of neurotrophic factors. This study was designed to investigate the effects of acute treatment of experimental closed head injury (CHI) in rats with Cerebrolysin on neurological function.

Methods

Adult male Wistar rats (n = 60) were subjected to impact acceleration–induced CHI. Closed head injured rats received intraperitoneal injection of saline (n = 30) or Cerebrolysin (2.5 ml/kg, n = 30) starting 1 hour postinjury and administered once daily until they were killed (2 or 14 days after CHI). To evaluate functional outcome, the modified neurological severity score (mNSS), foot fault, adhesive removal, and Morris water maze (MWM) tests were performed. Animals were killed on Day 14 (n = 20) after injury, and their brains were removed and processed for measurement of neuronal cells, axonal damage, apoptosis, and neuroblasts. The remaining rats (n = 40) were killed 2 days postinjury to evaluate cerebral microvascular patency by fluorescein isothiocyanate (FITC)–dextran perfusion (n = 16) and to measure the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase–9 (MMP-9) by using real-time reverse transcriptase-polymerase chain reaction (RT-PCR, n = 8) and by immunohistochemical analysis (n = 16).

Results

At 14 days post-CHI, the Cerebrolysin treatment group exhibited significant improvements in functional outcomes (the adhesive removal, mNSS, foot-fault, and MWM tests), and significantly more neurons and neuroblasts were present in the dentate gyrus (DG) (p < 0.05) compared with the saline-treated group (p < 0.05). At 2 days post-CHI, the Cerebrolysin group exhibited a significantly higher percentage of phosphorylated neurofilament H (pNF-H)–positive staining area in the striatum (p < 0.05), a significant increase in the percentage of FITC-dextran perfused vessels in the brain cortex (p < 0.05), a significant increase in the number of VEGF-positive cells (p < 0.05), and a significant reduction in the MMP-9 staining area (p < 0.05) compared with the saline-treated group. There was no significant difference in mRNA levels of MMP-9 and VEGF in the hippocampus and cortex 48 hours postinjury between Cerebrolysin- and saline-treated rats that sustained CHI.

Conclusions

Acute Cerebrolysin treatment improves functional recovery in rats after CHI. Cerebrolysin is neuroprotective for CHI (increased neurons in the dentate gyrus and the CA3 regions of the hippocampus and increased neuroblasts in the dentate gyrus) and may preserve axonal integrity in the striatum (significantly increased percentage of pNF-H–positive tissue in the striatum). Reduction of MMP-9 and elevation of VEGF likely contribute to enhancement of vascular patency and integrity as well as neuronal survival induced by Cerebrolysin. These promising results suggest that Cerebrolysin may be a useful treatment in improving the recovery of patients with CHI.

Abbreviations used in this paper:Ang1 = angiopoietin 1; BBB = blood-brain barrier; BDNF = brain-derived neurotrophic factor; CHI = closed head injury; CTNF = ciliary neurotrophic factor; DG = dentate gyrus; FITC = fluorescein isothiocyanate; GDNF = glial cell–derived neurotrophic factor; IGF = insulin-like growth factor; miRNA = microRNA; MMP-9 = matrix metalloproteinase–9; mNSS = modified Neurological Severity Score; MWM = Morris water maze; pNF-H = phosphorylated neurofilament H; RT-PCR = reverse transcriptase-polymerase chain reaction; SGZ = subgranular zone; TBI = traumatic brain injury; TUNEL = terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick-end labeling; VEGF = vascular endothelial growth factor.

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Contributor Notes

Address correspondence to: Ye Xiong, M.D., Ph.D., Department of Neurosurgery, Henry Ford Health System, E&R Building, Room #3096, 2799 West Grand Boulevard, Detroit, Michigan 48202. email: yxiong1@hfhs.org.

Please include this information when citing this paper: published online April 12, 2013; DOI: 10.3171/2013.3.JNS122061.

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