title: Tetramethylpyrazine Protects Against Oxygen-Glucose Deprivation-Induced Brain Microvascular Endothelial Cells Injury via Rho/Rho-kinase Signaling Pathway.
abstract: 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.
authors.affiliation: Key Laboratory of Xin'an Medicine, Ministry of Education, Hefei, 230012, China.|Anhui University of Chinese Medicine, Qian Jiang Road 1, Hefei, 230012, China.|Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine, Anhui Academy of Chinese Medicine, Hefei, 230012, China.
authors.forename: Guang
authors.initials: G
authors.lastname: Yang
authors.affiliation: Key Laboratory of Xin'an Medicine, Ministry of Education, Hefei, 230012, China.|Anhui University of Chinese Medicine, Qian Jiang Road 1, Hefei, 230012, China.|Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine, Anhui Academy of Chinese Medicine, Hefei, 230012, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Qian
authors.affiliation: Key Laboratory of Xin'an Medicine, Ministry of Education, Hefei, 230012, China. wnhefei@163.com.|Anhui University of Chinese Medicine, Qian Jiang Road 1, Hefei, 230012, China. wnhefei@163.com.|Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine, Anhui Academy of Chinese Medicine, Hefei, 230012, China. wnhefei@163.com.
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.affiliation: Key Laboratory of Xin'an Medicine, Ministry of Education, Hefei, 230012, China.|Anhui University of Chinese Medicine, Qian Jiang Road 1, Hefei, 230012, China.|Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine, Anhui Academy of Chinese Medicine, Hefei, 230012, China.
authors.forename: Chenyu
authors.initials: C
authors.lastname: Lin
authors.affiliation: Key Laboratory of Xin'an Medicine, Ministry of Education, Hefei, 230012, China.|Anhui University of Chinese Medicine, Qian Jiang Road 1, Hefei, 230012, China.|Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine, Anhui Academy of Chinese Medicine, Hefei, 230012, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory of Xin'an Medicine, Ministry of Education, Hefei, 230012, China.|Anhui University of Chinese Medicine, Qian Jiang Road 1, Hefei, 230012, China.|Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine, Anhui Academy of Chinese Medicine, Hefei, 230012, China.
authors.forename: Guangyun
authors.initials: G
authors.lastname: Wang
authors.affiliation: Key Laboratory of Xin'an Medicine, Ministry of Education, Hefei, 230012, China.|Anhui University of Chinese Medicine, Qian Jiang Road 1, Hefei, 230012, China.|Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine, Anhui Academy of Chinese Medicine, Hefei, 230012, China.
authors.forename: Xinxin
authors.initials: X
authors.lastname: Piao
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000187:drug effects* / Q000378:metabolism; D001921:Brain / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D008297:Male; D055806:Microvessels / Q000187:drug effects* / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D020011:Protective Agents / Q000494:pharmacology; D011719:Pyrazines / Q000494:pharmacology*; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects*; D054460:rho-Associated Kinases / Q000378:metabolism*
chemical_list: D004365:Drugs, Chinese Herbal; D020011:Protective Agents; D011719:Pyrazines; D052250:Nitric Oxide Synthase Type III; D054460:rho-Associated Kinases; D005947:Glucose; D010100:Oxygen; C017953:tetramethylpyrazine
keywords: Brain microvascular endothelial cells (BMECs); Oxygen-glucose deprivation (OGD); Rho/ROCK signaling pathway; Tetramethylpyrazine (TMP)