J Korean Neurosurg Soc.  2018 Sep;61(5):548-558. 10.3340/jkns.2017.0200.

S100ß, Matrix Metalloproteinase-9, D-dimer, and Heat Shock Protein 70 Are Serologic Biomarkers of Acute Cerebral Infarction in a Mouse Model of Transient MCA Occlusion

Affiliations
  • 1Department of Neurosurgery, Hallym University Kangnam Sacred Heart Hospital, Hallym University College of Medicine, Seoul, Korea.
  • 2Department of Neurosurgery, Korea University Medical Center, Seoul, Korea. sungkha@yahoo.com

Abstract


OBJECTIVE
Diagnosing acute cerebral infarction is crucial in determining prognosis of stroke patients. Although many serologic tests for prompt diagnosis are available, the clinical application of serologic tests is currently limited. We investigated whether S100β, matrix metalloproteinase-9 (MMP-9), D-dimer, and heat shock protein 70 (HSP70) can be used as biomarkers for acute cerebral infarction.
METHODS
Focal cerebral ischemia was induced using the modified intraluminal filament technique. Mice were randomly assigned to 30-minute occlusion (n=10), 60-minute occlusion (n=10), or sham (n=5) groups. Four hours later, neurological deficits were evaluated and blood samples were obtained. Infarction volumes were calculated and plasma S100β, MMP-9, D-dimer, and HSP70 levels were measured using enzyme-linked immunosorbent assay.
RESULTS
The average infarction volume was 12.32±2.31 mm³ and 46.9±7.43 mm³ in the 30- and 60-minute groups, respectively. The mean neurological score in the two ischemic groups was 1.6±0.55 and 3.2±0.70, respectively. S100β, MMP-9, and HSP70 expressions significantly increased after 4 hours of ischemia (p=0.001). Furthermore, S100β and MMP-9 expressions correlated with infarction volumes (p < 0.001) and neurological deficits (p < 0.001). There was no significant difference in D-dimer expression between groups (p=0.843). The area under the receiver operating characteristic curve (AUC) showed high sensitivity and specificity for MMP-9, HSP70 (AUC=1), and S100β (AUC=0.98).
CONCLUSION
S100β, MMP-9, and HSP70 can complement current diagnostic tools to assess cerebral infarction, suggesting their use as potential biomarkers for acute cerebral infarction.

Keyword

Acute ischemic stroke; Biomarkers; Cerebral infarction volume

MeSH Terms

Animals
Biomarkers*
Brain Ischemia
Cerebral Infarction*
Complement System Proteins
Diagnosis
Enzyme-Linked Immunosorbent Assay
Heat-Shock Proteins*
Hot Temperature*
HSP70 Heat-Shock Proteins*
Humans
Infarction
Ischemia
Matrix Metalloproteinase 9*
Mice*
Plasma
Prognosis
ROC Curve
Sensitivity and Specificity
Serologic Tests
Stroke
Biomarkers
Complement System Proteins
HSP70 Heat-Shock Proteins
Heat-Shock Proteins
Matrix Metalloproteinase 9

Figure

  • Fig. 1. A : coronal brain sections. Infarcts are visualized as unstained regions following 2, 3, 5-triphenyltetrazolium chloride staining. No infarct is seen in sham-operated mice. Infarcts are confined to the basal ganglia in 30-minute McAO mice, and extend from the basal ganglia to the cerebral cortex in the 60-minute McAO mice. b : comparison of the infarction volumes between the sham-operated, 30-minute McAO, and 60-minute McAO groups. The infarction volume is significantly larger in the 60-minute McAO group when compared with the 30-minute group (p=0.008). Data are expressed as mean±standard deviation. *p<0.05. McAO : middle cerebral artery occlusion.

  • Fig. 2. comparison of the neurological scores between the shamoperated, 30-minute McAO, and 60-minute McAO groups. Shamoperated mice do not show any neurological deficit. The neurological score is significantly higher in the 60-minute group when compared with the 30-minute group (p=0.032). Data are expressed as mean±standard deviation. *p<0.05. McAO : middle cerebral artery occlusion.

  • Fig. 3. Plasma S100β concentration in all groups. The concentration of S100β increases with occlusion time (p=0.001; A) and correlates significantly with infarct volume (r=0.867, p<0.001; b) and neurological deficit (r=0.802, p<0.001; c). *p<0.05.

  • Fig. 4. Plasma MMP-9 concentration in all groups. The concentration of MMP-9 increases with occlusion time (p=0.001; A) and correlates significantly with infarct volume (r=0.890, p<0.001; b) and neurological deficit (r=0.879, p<0.001; c). *p<0.05. MMP-9 : matrix metalloproteinase-9.

  • Fig. 5. Plasma D-dimer concentration in all groups. There is no significant difference in D-dimer concentration between groups (p=0.843; A), and no correlation with infarct volume (r=0.255, p=0.218; b) or neurological score (r=0.139, p=0.507; c). McAO : middle cerebral artery occlusion.

  • Fig. 6. Plasma HSP70 concentration in all groups. HSP70 concentration increases significantly in both infarction groups (p=0.001) when compared to that of the sham group (A); however, there is no correlation between HSP70 concentration and infarct area (r=0.174, p=0.406; b) or neurological score (r=0.138, p=0.510; c). *p<0.05. HSP70 : heat shock protein 70, McAO : middle cerebral artery occlusion.

  • Fig. 7. Sensitivity and specificity of the biomarkers. MMP-9 and HSP70 both have an AUc of 1. S100β has an AUc of 0.98. The AUc for D-dimer is 0.58. MMP-9 : matrix metalloproteinase-9, HSP70 : heat shock protein 70, AUc : the area under the receiver operating characteristic curve.


Reference

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