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Delayed Docosahexaenoic Acid Treatment Combined with Dietary Supplementation of Omega-3 Fatty Acids Promotes Long-Term Neurovascular Restoration After Ischemic Stroke

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Abstract

Prophylactic dietary intake of omega-3 polyunsaturated fatty acids (n-3 PUFAs) has been shown to remarkably ameliorate ischemic brain injury. However, the therapeutic efficacy of n-3 PUFA administration post-stroke, especially its impact on neurovascular remodeling and long-term neurological recovery, has not been fully characterized thus far. In this study, we investigated the effect of n-3 PUFA supplementation, as well as in combination with docosahexaenoic acid (DHA) injections, on long-term stroke outcomes. Mice were subjected to transient middle cerebral artery occlusion (MCAO) before randomly assigned to four groups to receive the following: (1) low dose of n-3 PUFAs as the vehicle control, (2) intraperitoneal DHA injections, (3) n-3 PUFA dietary supplement, or (4) combined treatment of (2) and (3). Neurological deficits and brain atrophy, neurogenesis, angiogenesis, and glial scar formation were assessed up to 28 days after MCAO. Results revealed that groups 2 and 3 showed only marginal reduction in post-stroke tissue loss and attenuation of cognitive deficits. Interestingly, group 4 exhibited significantly reduced tissue atrophy and improved cognitive functions compared to groups 2 and 3 with just a single treatment. Mechanistically, the combined treatment promoted post-stroke neurogenesis and angiogenesis, as well as reduced glial scar formation, all of which significantly correlated with the improved spatial memory in the Morris water maze. These results demonstrate an effective therapeutic regimen to enhance neurovascular restoration and long-term cognitive recovery in the mouse model of MCAO. Combined post-stroke DHA treatment and n-3 PUFA dietary supplementation thus may be a potential clinically translatable therapy for stroke or related brain disorders.

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Acknowledgments

*H.P. and *X.J. contributed equally to this research. This project was supported by the US Department of Veterans Affairs (VA) RR&D Merit Review RX000420; the US National Institutes of Health grants NS045048, NS091175, and NS095671; the American Heart Association grant 13SDG14570025; and the Chinese Natural Science Foundation grants 81529002, 81171149, 81371306, 81571285, and 81100978. J.C. is a recipient of the VA Senior Research Career Scientist Award. The authors are indebted to Pat Strickler for excellent administrative support. The present address of J.X. is Cerebrovascular Center, Henan Provincial People’s Hospital, Zhengzhou University, Zhengzhou 450003, China.

Authors’ Contributions

Y.S., Y.G., X.H., and J.C. designed the research. H.P., X.J., J.X., and W.Z. performed the research. X.J. and Y.S. analyzed the data. H.P., X.J., D.H., J.C., and Y.S. wrote the manuscript. All authors reviewed and edited the manuscript.

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Correspondence to Jun Chen or Yejie Shi.

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All animal procedures were approved by the University of Pittsburgh Institutional Animal Care and Use Committee and performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Hongjian Pu and Xiaoyan Jiang contributed equally to this work.

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Supplementary Fig. 1

Post-stroke DHA and FO treatments do not change hippocampal neurogenesis in the non-injured contralateral hemisphere. a Representative images of double-label immunostaining of BrdU (green) and NeuN (red) in the hippocampal CA1, CA3 and DG areas of the non-injured contralateral hemisphere at 28 days after MCAO. There was barely any BrdU immunosignal in all regions. Scale bar: 50 μm. b-d Quantification of total cells that are positive for NeuN immunosignal in CA1, CA3, and DG of the contralateral hippocampus. There was no statistical difference among all groups. e-g Quantification of BrdU+/NeuN+ cells in CA1, CA3, and DG of the contralateral hippocampus. Few BrdU+/NeuN+ cells were observed in all regions. There was no statistical difference among all groups. n = 7 mice per group. (DOCX 2039 kb)

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Pu, H., Jiang, X., Hu, X. et al. Delayed Docosahexaenoic Acid Treatment Combined with Dietary Supplementation of Omega-3 Fatty Acids Promotes Long-Term Neurovascular Restoration After Ischemic Stroke. Transl. Stroke Res. 7, 521–534 (2016). https://doi.org/10.1007/s12975-016-0498-y

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