CaMKII and MEK1/2 inhibition time-dependently modify inflammatory signaling in rat cerebral arteries during organ culture

Journal of Neuroinflammation

Lars Edvinsson¹  Leif E Johnson²  Hilda Ahnstedt¹  Sajedeh Eftekhari¹  Roya Waldsee¹ 

[1]Department of Clinical Sciences, Division of Experimental Vascular Research, Lund University, Sölvegatan 17, SE-221 84 Lund, Sweden

[2]Department of Ophthalmology, Division of Clinical Science, Lund University, Klinikgatan 26, SE- 221 85 Lund, Sweden

关键词: p38;    JNK;    Inflammation;    Organ culture;    CaMKII;    ERK1/2;   

Others  :  804168
DOI  :  10.1186/1742-2094-11-90

 received in 2013-10-23, accepted in 2014-05-01,  发布年份 2014

【 摘 要 】

Background

Cerebral ischemia induces transcriptional upregulation of inflammatory genes in the brain parenchyma and in cerebral arteries, thereby contributing to the infarct development. The present study was designed to evaluate the involvement of calcium-calmodulin-dependent protein kinase (CaMKII) II and extracellular signal-regulated kinase1/2 (ERK1/2) on inflammatory mediators in rat cerebral arteries using organ culture as a method for inducing ischemic-like vascular wall changes.

Methods

Rat basilar arteries were cultured in serum-free medium for 0, 3, 6 or 24 hours in the presence or absence of the CaMKII inhibitor KN93 or the MEK1/2 inhibitor U0126. Protein expression of activated CaMKII, ERK1/2, and inflammatory-associated protein kinases and mediators were examined with western blot and immunohistochemistry. Caspase-3 mRNA levels in basilar arteries were studied with real-time PCR.

Results

Western blot evaluation showed that organ culture induced a significant increase in phosphorylated ERK1/2 at 3, 6 and 24 hours, while CaMKII was found to be already activated in fresh non-incubated arteries and to decrease with incubation time. The addition of U0126 or KN93 decreased levels of phosphorylated c-Jun N-terminal kinase and p-p38, as evaluated by immunohistochemistry. KN93 affected the increase in caspase-3 mRNA expression only when given at the start of incubation, while U0126 had an inhibitory effect when given up to six hours later. Tumor necrosis factor receptor 1 was elevated after organ culture. This inflammatory marker was reduced by both of the two different protein kinase inhibitors.

Conclusions

The novel findings of the present study are that the cross-talk between the two protein kinases and the inhibition of CaMKII or MEK1/2 in a time-dependent manner attenuates inflammatory-associated protein kinases and mediators, suggesting that they play a role in cerebrovascular inflammation.

【 授权许可】

   
2014 Waldsee et al.; licensee BioMed Central Ltd.

【 图 表 】

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