Identification of potential biomarkers for artificial cold exposure-induced hypertensive stroke by proteomic analysis

J Stroke Cerebrovasc Dis. 2014 Nov-Dec;23(10):2671-2680. doi: 10.1016/j.jstrokecerebrovasdis.2014.06.014. Epub 2014 Oct 11.


Background: The effect of changing temperature on an individual's cerebrovascular risk is both biologically plausible and supported by epidemiologic evidence. We used a global proteomic-based approach to analyze the expression alterations of proteins in artificial cold exposure (ACE)-induced hypertensive stroke in renovascular hypertensive rats (RHR) and to identify the biomarker of ACE-induced hypertensive stroke.

Methods: The RHR models were established by 2 kidney 2 clip methods. ACE treatment was achieved using an intelligent artificial climate cabinet. Blood pressure and neurologic symptoms were observed before and after ACE treatment. Hemorrhagic condition and infarction survey were examined using 2,3,5-triphenyltetrazolium chloride staining. The total number of proteins derived from the cerebral tissue of the RHR models were analyzed with 2-dimensional gel electrophoresis (2-DE), ImageMaster 2D Platinum software, and mass spectrometry. Significantly regulated proteins selected for further functional studies using the Search Tool for the Retrieval of Interacting Genes/Proteins system were verified by Western blot.

Results: ACE-induced stroke in the RHR group (31.25%, 25 of 80 vs. 16.25%, 13 of 80; P < .05) but not in the sham-operated group. Following ACE treatment, we identified 37 differentially expressed proteins and 28 were unique. Two of the upregulated proteins, Syt1 and Idh3a, were obtained by bioinformatics analysis and verified by Western blot.

Conclusions: The rate of morbidity as a result of stroke in RHR was obviously elevated after ACE treatment. ACE might affect protein expression profile in cerebral tissues of RHR. Syt1 and Idh3a may play a vital role in ACE-induced hypertensive stroke.

Keywords: Artificial cold exposure; Idh3a; Syt1; hypertensive stroke.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / metabolism*
  • Blood Pressure
  • Blotting, Western
  • Cold Temperature / adverse effects*
  • Disease Models, Animal
  • Hypertension, Renal / complications*
  • Isocitrate Dehydrogenase / metabolism
  • Kidney / blood supply*
  • Kidney / surgery
  • Male
  • Proteomics
  • Rats
  • Renal Circulation*
  • Stroke / etiology
  • Stroke / metabolism*
  • Synaptotagmin I / metabolism
  • Tetrazolium Salts / chemistry


  • Biomarkers
  • Synaptotagmin I
  • Syt1 protein, rat
  • Tetrazolium Salts
  • triphenyltetrazolium
  • Isocitrate Dehydrogenase