Cellular and Molecular Neurobiology

, Volume 39, Issue 2, pp 301–319 | Cite as

Myosin IIA Regulated Tight Junction in Oxygen Glucose-Deprived Brain Endothelial Cells Via Activation of TLR4/PI3K/Akt/JNK1/2/14-3-3ε/NF-κB/MMP9 Signal Transduction Pathway

  • Yanni LvEmail author
  • Wen Liu
  • Zhaohui Ruan
  • Zixuan Xu
  • Longsheng Fu
Original Research


Non-muscle myosin heavy chain IIA (NMMHC IIA), a member of Myosin II family, plays a critical role in various cellular physiological processes. Our previous research had suggested that NMMHC IIA could participate in regulating tight junction morphological changes induced by ischemia stroke. Thus, in the current study, we attempted to uncover the regulation pattern of NMMHC IIA on tight junction dysfunction in oxygen glucose-deprived (OGD) mouse brain bEND.3 endothelial cells. The regulation of NMMHC IIA on tight junction in OGD-stimulated bEND.3 cells was evaluated by western blotting assay. Morphologic change of occludin, claudin-5, and ZO-1 tight junction proteins was compared with pretreatment with NMMHC II inhibitor blebbistatin via immunohistochemical staining. Detection of activation of NMMHC IIA on OGD-mediated tight junction transduction pathway was investigated via Koch’s postulate using corresponding protein inhibitor. Our results showed that NMMHC IIA was activated in OGD-stimulated bEND.3 endothelial cells. The inhibition of NMMHC IIA could attenuate the morphologic change of occludin, claudin-5, and ZO-1 tight junction proteins. NMMHC IIA participated in regulating downstream transduction pathway TLR4, phosphatidylinositol 3-kinase (PI3K), Akt, JNK1/2, 14-3-3ε, nuclear factor kappa B (NF-кB) and matrix metalloprotein 9 (MMP9). Blocking of these pathways using indicated inhibitors demonstrated that NMMHC IIA destroyed the connection of tight junction via the activation of TLR4/PI3K/Akt/JNK1/2/14-3-3ε/NF-κB/MMP9 pathway. Our study described the key role of NMMHC IIA in OGD-stimulated mouse brain bEND.3 endothelial cells, while also exhibited the molecule effect on tight junction dysfunction via TLR4/PI3K/Akt/JNK1/2/14-3-3ε/NF-κB/MMP9 signal transduction pathway.


Non-muscle myosin heavy chain IIA Mouse brain bEND.3 endothelial cells Tight junction dysfunction TLR4/PI3K/Akt/JNK1/2/14-3-3ε/NF-κ/BMMP9 



This work was supported by Foundation Project: National Natural Science Foundation of China (81760094, 31602111, 81860020), China; Jiangxi Provincial Education Department Science and Technology Research Youth Project (No. GJJ160238); The Foundation of Jiangxi Provincial Department of Science and Technology Youth Project (No. 20171BAB215021). The funder has no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript.

Author Contributions

All authors contributed to the research. YL conceived the experiments. YL, WL, ZR, ZX, LF conducted the experiments and analyzed the results. All authors contributed to the writing and editing of the manuscript.

Compliance with Ethical Standards

Conflict of interest

None of the authors have any conflicts of interest, including financial, personal or other relationships, with other individuals or organizations.

Ethical Approval

All animal care and experimental procedures were conducted according to standard ethical guidelines (National Institutes of Health Guide to the use of Laboratory Animals) and approved by Institutional Animal Care and use Committee of Nanchang University. All efforts were made to minimize the number of mice used and their suffering.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Pharmacy departmentThe First Affiliated Hospital of Nanchang UniversityNanchangChina
  2. 2.Jiangxi Provincial Institute of Traditional Chinese MedicineNanchangChina

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