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Hemoglobin-Induced Nitric Oxide Synthase Overexpression and Nitric Oxide Production Contribute to Blood–Brain Barrier Disruption in the Rat

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Abstract

Hemoglobin (Hb) released from extravasated erythrocytes may have a critical role in the process of blood–brain barrier (BBB) disruption and subsequent edema formation after intracerebral hemorrhage (ICH). Excessive nitric oxide (NO) production synthesized by nitric oxide synthase (NOS) has been well documented to contribute to BBB disruption. However, considerably less attention has been focused on the role of NO in Hb-induced BBB disruption. This study was designed to examine the hypothesis that Hb-induced NOS overexpression and excessive NO production may contribute to the changes of tight junction (TJ) proteins and subsequent BBB dysfunction. Hemoglobin was infused with stereotactic guidance into the right caudate nucleus of male Sprague Dawley rats. Then, we investigated the effect of Hb on the BBB permeability, changes of TJ proteins (claudin-5, occludin, zonula occludens-1 (ZO-1), and junctional adhesion molecule-1 (JAM-1)), iron deposition, expression of inducible NOS (iNOS) and endothelial NOS (eNOS), as well as NO production. Hb injection caused a significant increase in BBB permeability. Significant reduction of claudin-5, ZO-1, and JAM-1 was observed after Hb injection as evidenced by PCR and immunofluorescence. After a decrease at early stage, occludin showed a fivefold increase in mRNA level at 7 days. Significant iron deposition was detectable from 48 h to 7 days in a time-dependent manner. The iNOS and eNOS levels dramatically increased after Hb injection concomitantly with large quantities of NO released. Furthermore, enhanced iNOS or eNOS immunoreactivity was co-localized with diffused or diminished claudin-5 staining. We concluded that overexpressed NOS and excessive NO production induced by Hb may contribute to BBB disruption, which may provide an important potential therapeutic target in the treatment of ICH.

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (no. 81271314), Natural Science Foundation of Guangdong (no. 5300468), and Special Project on the Integration of Industry, Education and Research of Guangdong Province and Ministry of Education (no. 2012B091100154).

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Authors and Affiliations

  1. The National Key Clinic Specialty, The Neurosurgery Institute of Guangdong Province, Guangdong Provincial Key Laboratory on Brain Function Repair and Regeneration, Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou, 510282, China

    Shuo Yang, Yizhao Chen, Xinqing Deng, Zhenghao Fu, Mouxuan Du & Rui Ding

  2. Department of Neurosurgery, The First Affiliated Hospital, University of South China, Hengyang, 421001, China

    Weiping Jiang

  3. Department of Neurosurgery, The First Affiliated Hospital of He’nan University of Science and Technology, Luoyang, 471003, China

    Bing Li

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  1. Shuo Yang
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Correspondence to Yizhao Chen.

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Yang, S., Chen, Y., Deng, X. et al. Hemoglobin-Induced Nitric Oxide Synthase Overexpression and Nitric Oxide Production Contribute to Blood–Brain Barrier Disruption in the Rat. J Mol Neurosci 51, 352–363 (2013). https://doi.org/10.1007/s12031-013-9990-y

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  • DOI: https://doi.org/10.1007/s12031-013-9990-y

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