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Doxycycline-Mediated Protective Effect Against Focal Cerebral Ischemia–Reperfusion Injury Through the Modulation of Tight Junctions and PKCδ Signaling in Rats

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Abstract

The strategy for the development of effective and safe neuroprotective agents has great potential to reduce cerebral ischemia–reperfusion injury and improve the functional outcome in stroke patients. Recently, doxycycline, a tetracycline antibiotic, has been shown to have neuroprotective efficiency in reduction of a variety of ischemia–reperfusion injuries as well as ischemic brain damage. We used the rat models of middle cerebral artery occlusion (MCAO) and reperfusion to investigate the effects of treatments with doxycycline against the blood-brain barrier (BBB) leakage at 3, 12, 72, and 120 h of reperfusion. Male Sprague–Dawley rats were subjected to MCAO for 2 h followed by reperfusion for 3, 12, 72, and 120 h and received either doxycycline (45 mg/kg) or saline. The results showed that the treatment of doxycycline significantly reduced the BBB leakage and cerebral infarct volume, which were proved by Evans blue assay and TTC staining. Real-time PCR, immunohistochemistry, and western blot assay verified that the administration of doxycycline significantly up-regulated the expression of tight junction claudin-5, occludin, and ZO-1 from 3 to 120 h after reperfusion. The results of real-time PCR, western blot, and gelatin zymography analyses revealed that the gene and protein expression and activities of matrix metalloproteinases (MMPs) MMP-2 and MMP-9 were significantly elevated in a different time-dependent manner after ischemia–reperfusion but significantly inhibited by doxycycline treatment. Moreover, doxycycline could also significantly down-regulate the expression of PKCδ mRNA and protein after ischemia–reperfusion. These results suggested that the protective effects of doxycycline against BBB damage induced by reperfusion might be related to the up-regulation of tight junction proteins and inhibition of MMP-2, MMP-9, and PKCδ.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (Nos. 30872656, 30973079, 81172197, 81171131, and 81072056), the special fund for Scientific Research of Doctor-degree Subjects in Colleges and Universities (Nos. 20092104110015 and 20102104110009), and Shenyang Science and Technology Plan Projects (Nos. F10-205-1-22 and F10-205-1-37).

Conflict of Interest Statement

The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Department of Physiology, College of Basic Medical Sciences, China Medical University, Shenyang, Liaoning Province, 110001, People’s Republic of China

    Zhenhua Wang

  2. Department of Neurobiology, College of Basic Medical Sciences, China Medical University, Shenyang, Liaoning Province, 110001, People’s Republic of China

    Yixue Xue, Haixia Jiao & Ping Wang

  3. Institute of Pathology and Pathophysiology, China Medical University, Shenyang, Liaoning Province, 110001, People’s Republic of China

    Yixue Xue, Haixia Jiao & Ping Wang

  4. Department of Neurosurgery, Shengjing Hospital, China Medical University, Shenyang, Liaoning Province, 110004, People’s Republic of China

    Yunhui Liu

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  1. Zhenhua Wang
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  2. Yixue Xue
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  3. Haixia Jiao
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  4. Yunhui Liu
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Correspondence to Yixue Xue.

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Zhenhua Wang, Yixue Xue, and Haixia Jiao contributed equally to this work.

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Wang, Z., Xue, Y., Jiao, H. et al. Doxycycline-Mediated Protective Effect Against Focal Cerebral Ischemia–Reperfusion Injury Through the Modulation of Tight Junctions and PKCδ Signaling in Rats. J Mol Neurosci 47, 89–100 (2012). https://doi.org/10.1007/s12031-011-9689-x

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  • DOI: https://doi.org/10.1007/s12031-011-9689-x

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