Cellular and Molecular Neurobiology

, Volume 37, Issue 4, pp 619–633 | Cite as

Tetramethylpyrazine Protects Against Oxygen-Glucose Deprivation-Induced Brain Microvascular Endothelial Cells Injury via Rho/Rho-kinase Signaling Pathway

  • Guang Yang
  • Chen Qian
  • Ning Wang
  • Chenyu Lin
  • Yan Wang
  • Guangyun Wang
  • Xinxin Piao
Original Research


Tetramethylpyrazine (TMP, also known as Ligustrazine), which is isolated from Chinese Herb Medicine Ligustium wollichii Franchat (Chuan Xiong), has been widely used in China for the treatment of ischemic stroke by Chinese herbalists. Brain microvascular endothelial cells (BMECs) are the integral parts of the blood–brain barrier (BBB), protecting BMECs against oxygen-glucose deprivation (OGD) which is important for the treatment of ischemic stroke. Here, we investigated the protective mechanisms of TMP, focusing on OGD-injured BMECs and the Rho/Rho-kinase (Rho-associated kinases, ROCK) signaling pathway. The model of OGD-injured BMECs was established in this study. BMECs were identified by von Willebrand factor III staining and exposed to fasudil, or TMP at different concentrations (14.3, 28.6, 57.3 µM) for 2 h before 24 h of OGD injury. The effect of each treatment was examined by cell viability assays, measurement of intracellular reactive oxygen species (ROS), and transendothelial electric resistance and western blot analysis (caspase-3, endothelial nitric oxide synthase (eNOS), RhoA, Rac1). Our results show that TMP significantly attenuated apoptosis and the permeability of BMECs induced by OGD. In addition, TMP could notably down-regulate the characteristic proteins in Rho/ROCK signaling pathway such as RhoA and Rac1, which triggered abnormal changes of eNOS and ROS, respectively. Altogether, our results show that TMP has a strong protective effect against OGD-induced BMECs injury and suggest that the mechanism might be related to the inhibition of the Rho/ROCK signaling pathway.


Tetramethylpyrazine (TMP) Oxygen-glucose deprivation (OGD) Brain microvascular endothelial cells (BMECs) Rho/ROCK signaling pathway 



The study was supported by the National Natural Science Foundation of China (Nos. 30973979, 81374005) and “Twelfth Five Year” National Science and Technology Support Program of China (No. 2012BAI26B03).

Compliance with Ethical Standards

Conflict of Interest


Ethical statement

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; (d) whole research process was 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.

Supplementary material

10571_2016_398_MOESM1_ESM.docx (15 kb)
Supplementary material 1 (DOCX 15 kb)


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Guang Yang
    • 1
    • 2
    • 3
  • Chen Qian
    • 1
    • 2
    • 3
  • Ning Wang
    • 1
    • 2
    • 3
  • Chenyu Lin
    • 1
    • 2
    • 3
  • Yan Wang
    • 1
    • 2
    • 3
  • Guangyun Wang
    • 1
    • 2
    • 3
  • Xinxin Piao
    • 1
    • 2
    • 3
  1. 1.Key Laboratory of Xin’an MedicineMinistry of EducationHefeiChina
  2. 2.Anhui University of Chinese MedicineHefeiChina
  3. 3.Institute for Pharmacodynamics and Safety Evaluation of Chinese MedicineAnhui Academy of Chinese MedicineHefeiChina

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