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Effects of Erythropoietin on Blood–Brain Barrier Tight Junctions in Ischemia–Reperfusion Rats

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Abstract

Administration of recombinant human erythropoietin (rhEPO) protects neurons from injury after brain ischemia–reperfusion (I/R), which is in part mediated by ameliorating the blood–brain barrier (BBB) leakage. But the mechanism of rhEPO’s protective effects on BBB remains unclear. This study aims to investigate the effects of rhEPO on BBB integrity and the expressions of tight junctions (TJs) associated proteins of zonula occluden-1 (ZO-1), occludin, and claudin-5 in cerebral I/R rats. These rats underwent 2 h of ischemia and then were reperfused for up to 3 and 72 h. Animals were randomly divided into five groups: sham-operated group, I/R 3 h and I/R 72 h group (2 ml saline was injected intraperitoneally just before the onset of ischemia), rhEPO + I/R 3 h, and rhEPO + I/R 72 h group (5,000 U/kg rhEPO diluted in 2 ml saline solution was injected intraperitoneally just before the onset of ischemia). We verified that rhEPO could decrease the BBB leakage induced by I/R injury detected by Evans blue extravasation. 2, 3, 5-Triphenyltetrazolium chloride staining results showed that rhEPO decreased infarct volume after cerebral I/R. TJ integrity was partly restored by rhEPO observed by transmission electron microscopy. The mRNA and protein expression levels of ZO-1, occludin, and claudin-5 were significantly increased compared with I/R groups at the same reperfusion time point by reverse transcriptase-polymerase chain reaction and Western blot assays. The treatment of rhEPO induced the redistribution of ZO-1, occludin, and claudin-5 in cerebral microvessels observed by immunohistochemical staining. Compared with I/R groups, the mRNA level of tumor necrosis factor-alpha (TNF-α) in cerebral microvessels decreased markedly after rhEPO treatment, accompanied with reduced TNF-α protein level and nuclear factor-кB (NF–кB) p65 activation detected by enzyme-linked immunosorbent assay. These results suggested that the protective mechanism of rhEPO on BBB after cerebral I/R injury was associated with the upregulation of TJ-associated proteins. The downregulated TNF-α levels and NF–кB activation induced by rhEPO might be involved in this process.

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Acknowledgements

This work is supported by grants from the Natural Science Foundation of China (nos. 81171131, 81172197, 30973079, 81072056, and 81100893), the special fund for Scientific Research of Doctor-degree Subjects in Colleges and Universities (nos. 20092104110015 and 20102104110009), the Natural Science Foundation of Liaoning Province in China (no. 201102300), Liaoning Science and Technology Plan Projects (no. 2011225020), and Shenyang Science and Technology Plan Projects (nos. F-10-205-1-22 and F-10-205-1-37).

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Authors and Affiliations

  1. Department of Neurobiology, College of Basic Medicine, China Medical University, Shenyang, Liaoning province, China, 110001

    Kun Liu, Ping Wang & Yi-xue Xue

  2. Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China, 100050

    Tao Sun & Li-wei Zhang

  3. Institute of Pathology and Pathophysiology, China Medical University, Shenyang, China, 110001

    Ping Wang & Yi-xue Xue

  4. Department of Neurosurgery, Shengjing Hospital, China Medical University, Shenyang, China, 110004

    Yun-hui Liu

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  1. Kun Liu
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  4. Yun-hui Liu
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Correspondence to Yi-xue Xue.

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Kun Liu, Tao Sun, and Ping Wang contributed equally to this work.

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Liu, K., Sun, T., Wang, P. et al. Effects of Erythropoietin on Blood–Brain Barrier Tight Junctions in Ischemia–Reperfusion Rats. J Mol Neurosci 49, 369–379 (2013). https://doi.org/10.1007/s12031-012-9883-5

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  • DOI: https://doi.org/10.1007/s12031-012-9883-5

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