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Effects of Muscone on the Expression of P-gp, MMP-9 on Blood–Brain Barrier Model In Vitro

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Abstract

Muscone is the main chemical ingredient in Musk which is main crude drug in Tongqiaohuoxue decoction (TQHXD), and TQHXD has a protective effect on damaged neurons, so we hypothesize that muscone can alter blood–brain barrier (BBB) permeability via the modulation of P-glycoprotein (P-gp) and matrix metalloproteinase-9 (MMP-9) expression. In this study, astrocytes (AC) and human umbilical vein endothelial cells (ECV304) were co-cultured to simulate the BBB model in vitro. Leak testing, transmembrane resistance experiments, and BBB-specific enzyme testing were used to test whether the model was successful. Different concentrations of muscone permeating the BBB were detected by gas chromatography (GC). The change of the transendothelial electrical resistance (TEER) on the BBB in vitro after treating with muscone was detected by Millicell-ERS. The protein expression of P-gp, MMP-9 in normal, and oxygen/glucose deprivation (OGD) BBB model was determined by western blotting to inquire that the mechanism of muscone penetrates the BBB model in vitro. The results show that muscone was detected in the lower medium of the BBB model by GC; the values of TEER were no significant difference before and after muscone (8 μM) was added to the BBB model; the expression of P-gp significantly decreased after the BBB model treatment with muscone (4, 8, and 16 μM) for 24 h; the expression of P-gp and MMP-9 in different concentrations of muscone groups had different degrees of reduction compared with the BBB in the state of OGD. In conclusion, muscone could permeate the BBB model, and it was associated with the inhibition of P-gp and MMP-9 expression. An understanding of the mechanisms of muscone across the BBB is crucial to the development of therapeutic modalities for cerebral vascular diseases.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant No. 81374005) and Outstanding Youth Science and Technology Fund of Anhui Province (Grant No. 10040606Y17).

Compliance with Ethical Standards

On behalf of, and having obtained permission from all the authors, I declare that (a) the material has not been published in whole or in part elsewhere; (b) the paper is not currently being considered for publication elsewhere; (c) all authors have been personally and actively involved in substantive work leading to the report, and will hold themselves jointly and individually responsible for its content; and (d) all protocols, including surgical procedures and animal use, were approved by the institutional Animal Care and Use Committee of the Anhui university of Chinese medicine and conformed to the Guide for the Care and Use of Laboratory Animals by the National Institutes of Health. I testify to the accuracy of the above on behalf of all the authors.

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Authors and Affiliations

  1. College of Pharmacy, Anhui University of Chinese Medicine, Qian Jiang Road, Hefei, 230012, China

    Guang-Yun Wang, Ning Wang & Hua-Ning Liao

  2. Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine, Anhui University of Chinese Medicine, Hefei, 230012, China

    Guang-Yun Wang, Ning Wang & Hua-Ning Liao

  3. Chinese Key Laboratory of Xin’an Medicine, Ministry of Education, Hefei, 230012, China

    Guang-Yun Wang, Ning Wang & Hua-Ning Liao

  4. Department of Pharmacy, Anhui University of Chinese Medicine, Xiang Shan Road, Hefei, China

    Ning Wang

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  1. Guang-Yun Wang
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Correspondence to Ning Wang.

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Wang, GY., Wang, N. & Liao, HN. Effects of Muscone on the Expression of P-gp, MMP-9 on Blood–Brain Barrier Model In Vitro. Cell Mol Neurobiol 35, 1105–1115 (2015). https://doi.org/10.1007/s10571-015-0204-8

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