MAPK signaling pathway regulates cerebrovascular receptor expression in human cerebral arteries.
BACKGROUND
Cerebral ischemia results in enhanced expression of contractile cerebrovascular receptors, such as endothelin type B (ET(B)), 5-hydroxytryptamine type 1B (5-HT(1B)), angiotensin II type 1 (AT(1)) and thromboxane (TP) receptors in the cerebral arteries within the ischemic area. The receptor upregulation occurs via activation of the mitogen-activated protein kinases (MAPK) pathway. Previous studies have shown that inhibitors of the MAPK pathway diminished the ischemic area and contractile cerebrovascular receptors after experimental cerebral ischemia. The aim of this study was to examine if the upregulation of contractile cerebrovascular receptors after 48 h of organ culture of human cerebral arteries involves MAPK pathways and if it can be prevented by a MEK1/2 inhibitor. Human cerebral arteries were obtained from patients undergoing intracranial tumor surgery. The vessels were divided into ring segments and incubated for 48 h in the presence or absence of the specific MEK1/2 inhibitor U0126. The vessels were then examined by using in vitro pharmacological methods and protein immunohistochemistry.


RESULTS
After organ culture of the cerebral arteries the contractile responses to endothelin (ET)-1, angiotensin (Ang) II and thromboxane (TP) were enhanced in comparison with fresh human arteries. However, 5-carboxamidotryptamine (5-CT) induced decreased contractile responses after organ culture as compared to fresh arteries. Incubation with U0126 diminished the maximum contraction elicited by application of ET-1, Ang II and U46619 in human cerebral arteries. In addition, the MEK1/2 inhibitor decreased the contractile response to 5-CT. Immunohistochemistry revealed that organ culture resulted in increased expression of endothelin ET(A), endothelin ET(B) angiotensin AT(2), 5-hydroxytryptamine 5-HT(1B) and thromboxane A2 receptors, and elevated levels of activated pERK1/2, all localized to the smooth muscle cells of the cerebral arteries. Co-incubation with U0126 normalized these proteins.


CONCLUSIONS
The study demonstrated that there is a clear association between human cerebrovascular receptor upregulation via transcription involving activation of the MAPK pathway after organ culture. Inhibition of the MAPK pathways attenuated the vasoconstriction mediated by ET, AT and TP receptors in human cerebral arteries and the enhanced expression of their receptors. The results indicate that MAPK inhibition might be a novel target for treatment of cerebrovascular disorders.
Department of Clinical Sciences, Division of Experimental Vascular Research, Lund University, Lund, Sweden. Saema.Ansar@med.lu.se
Saema
S
Ansar
Sajedeh
S
Eftekhari
Roya
R
Waldsee
Elisabeth
E
Nilsson
Ola
O
Nilsson
Hans
H
Säveland
Lars
L
Edvinsson
D000804:Angiotensin II / Q000378:metabolism; D002070:Butadienes / Q000494:pharmacology; D002536:Cerebral Arteries / Q000187:drug effects / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D019332:Endothelin-1 / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D009119:Muscle Contraction / Q000187:drug effects; D009132:Muscles / Q000187:drug effects; D009213:Myography; D009570:Nitriles / Q000494:pharmacology; D009924:Organ Culture Techniques; D044022:Receptor, Endothelin A / Q000378:metabolism; D044023:Receptor, Endothelin B / Q000378:metabolism; D044302:Receptor, Serotonin, 5-HT1B / Q000378:metabolism; D011945:Receptors, Angiotensin / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017482:Receptors, Thromboxane / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*
D002070:Butadienes; D019332:Endothelin-1; D004791:Enzyme Inhibitors; D009570:Nitriles; D044022:Receptor, Endothelin A; D044023:Receptor, Endothelin B; D044302:Receptor, Serotonin, 5-HT1B; D011945:Receptors, Angiotensin; D011956:Receptors, Cell Surface; D017482:Receptors, Thromboxane; C113580:U 0126; D000804:Angiotensin II; D020928:Mitogen-Activated Protein Kinases