Gene expression and molecular changes in cerebral arteries following subarachnoid hemorrhage in the rat

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Object

The authors investigated early changes in the cerebral arteries of rats that occur after subarachnoid hemorrhage (SAH).

Methods

Messenger RNA was investigated by performing microarray and quantitative real-time polymerase chain reaction (PCR) analyses, and protein expression was shown by performing immunohistochemical studies. The array data indicated that the initial processes that occur after SAH involve activation of genes involved in angiogenesis, inflammation, and extracellular matrix (ECM) remodeling. The real-time PCR investigation confirmed upregulation of genes that were observed using the microarray to be regulated, including iNOS, MMP13, and cxcl2. The authors also verified the upregulation of previously implicated genes for G-protein–coupled receptors (endothelin B [ETB], angiotensin 1 [AT1], and AT2) and metalloproteinase 9. The results of an immunohistochemical study confirmed that receptor genes that were seen to be regulated produced an increase in protein expression. Double immunostaining of rat cerebral arteries with endothelial cell– or smooth-muscle cell–specific antibodies verified that an increase in ETB, 5-hydrotryptamine (5-HT1B), and 5-HT1D receptor expression occurs in smooth-muscle cells.

Conclusions

Processes occurring after SAH lead to enhanced arterial contractility and ECM remodeling either directly or through angiogenesis and inflammation. These processes are active via an increase in metalloproteinase expression, the presence of proangiogenic factors, and the expression of proinflammatory genes.

Abbreviations used in this paper: AT = angiotensin receptor; BA = basilar artery; CBF = cerebral blood flow; cDNA = complementary DNA; EASE = Expression Analysis Systematic Explorer; ECM = extracellular matrix; ET = endothelin; ICP = intracranial pressure; iNOS = inducible nitric oxide synthase; MCA = middle cerebral artery; MMP = matrix metalloproteinase; mRNA = messenger RNA; OD = optical density; OMIM = Online Mendelian Inheritance in Man; PCR = polymerase chain reaction; SAH = subarachnoid hemorrhage; TNF = tumor necrosis factor-α; VEGF = vascular endothelial growth factor; 5-HT = 5-hydrotryptamine.
Article Information

Contributor Notes

Address reprint requests to: Petter Vikman, Ph.D., Department of Clinical Sciences, Experimental Vascular Research, 3MC A13, S221 84, Lunds Universitet, Lund, Sweden. email: Petter.vikman@med.lu.se.

© AANS, except where prohibited by US copyright law.

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