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Ramelteon Ameliorates LPS-Induced Hyperpermeability of the Blood-Brain Barrier (BBB) by Activating Nrf2

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Abstract

The blood-brain barrier (BBB) is important for protecting the brain tissue by selectively exchanging substances between the blood and brain. The integrity of the BBB can be damaged by multiple factors, including oxidative stress and inflammation. Ramelteon is an oral hypnotic drug, and in the present study, we investigated its protective effect on BBB damage, as well as the underlying mechanism. LPS was used to induce BBB damage on mice and stimulate injury on endothelial cells. Evans blue staining assay was used to measure the brain permeability. The expressions of ZO-1 and Occludin were evaluated using immunostaining and Western blot in the brain tissue and endothelial cells, respectively. qRT-PCR and ELISA were used to detect the production of IL-1β and MCP-1 in the brain vessels. TBA assay was utilized to examine the concentration of MDA in the brain tissue and endothelial cells. The expression of Nrf2 in the nucleus and NQO1 were determined using Western blot assay. The endothelial permeability of the monolayer was examined using the FITC-dextran permeation assay. Firstly, the increased brain permeability and downregulated expression of tight junction proteins in the brain tissue induced by LPS were significantly reversed by treatment with Ramelteon, accompanied by the decrease in the production of IL-1β and MCP-1 in the vessels in mice. Also, the Nrf2 signaling was activated and oxidative stress in the brain vessels was alleviated by treatment with Ramelteon. Secondly, LPS-induced increase in endothelial monolayer permeability and decrease in tight junction protein expression in bEnd.3 brain endothelial cells were significantly reversed by Ramelteon, accompanied by activated Nrf2 signaling and alleviated oxidative stress. Lastly, the protective effects of Ramelteon against LPS-induced reduction of ZO-1 and Occludin, and the increase in endothelial monolayer permeability were dramatically abolished by silencing Nrf2. Ramelteon might ameliorate LPS-induced hyperpermeability of the BBB by activating the Nrf2 signaling pathway.

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Data availability

The data that support the findings of this study are available on request from the corresponding author.

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Funding

This study is funded by the Department of Health of Yunnan Province (YNH-20180056).

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Authors and Affiliations

  1. Department of NeurologyI, Clinical Medical College of DaLi University, No.32, Jiashibo Avenue, Dali Bai Autonomous Prefecture, 671000, Yunnan, China

    Yonglei Liu, Lixia Wang, Ning Du, Xiaoling Yin, Hongtao Shao & Lin Yang

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  1. Yonglei Liu
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  2. Lixia Wang
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  3. Ning Du
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  4. Xiaoling Yin
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  5. Hongtao Shao
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  6. Lin Yang
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Contributions

Yonglei Liu and Lin Yang designed the experiments and analyzed the data; Yonglei Liu, Lixia Wang, Ning Du, Xiaoling Yin, and Hongtao Shao performed the experiments and collected the results; Lin Yang prepared the manuscript. All authors have read and approved the final version.

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Correspondence to Lin Yang.

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Animal experiment protocols were approved by the Laboratory Animal Care Committee of DaLi University.

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Liu, Y., Wang, L., Du, N. et al. Ramelteon Ameliorates LPS-Induced Hyperpermeability of the Blood-Brain Barrier (BBB) by Activating Nrf2. Inflammation 44, 1750–1761 (2021). https://doi.org/10.1007/s10753-021-01451-w

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  • DOI: https://doi.org/10.1007/s10753-021-01451-w

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