title: YiQiFuMai Powder Injection ameliorates the oxygen-glucose deprivation-induced brain microvascular endothelial barrier dysfunction associated with the NF-κB and ROCK1/MLC signaling pathways.
abstract: BACKGROUND
  YiQiFuMai Powder Injection (YQFM) is a modern preparation derived from Sheng-mai San, a traditional Chinese prescription, widely used for the treatment of cardiovascular and cerebrovascular diseases. However, its potential molecular mechanism remains unclear.
  
  
  OBJECTIVE
  The present study was designed to observe the effect of YQFM on oxygen-glucose deprivation (OGD)-induced the brain microvascular endothelial barrier dysfunction and to explore the underlying pathways in vitro.
  
  
  METHODS
  A mouse brain microvascular endothelial cell line (bEnd.3) was subjected to OGD (2-9h) to examine the efficacy and molecular mechanisms in the presence or absence of YQFM (100, 200 and 400 μg/ml).
  
  
  RESULTS
  The results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Trans-endothelial electrical resistance (TEER) assays demonstrated that treatment with YQFM increased the cell viability and TEER value, decreased even blue (EB) albumin leakage after OGD in bEnd.3 cells. Western blotting and immunofluorescence staining showed that YQFM reduced the breakage and translocation of Zonula occludens-1 (ZO-1) and claudin-5 after 4h of OGD and decreased the expression of these proteins after 9h of OGD. Moreover, YQFM significantly inhibited the expression, phosphorylation and nuclear translocation of NF-κB/p65 and decreased the expression of intercellular adhesionmolecule-1 (ICAM-1) and cyclooxygenase (COX-2) as well as production of nitric oxide (NO). In addition, real time-PCR results revealed that YQFM suppressed the mRNA levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) after 4h of OGD. Furthermore, YQFM markedly inhibited both the phosphorylation of myosin light chain (MLC) and cytoskeletal reorganization and reduced the expression of cleaved-ROCK1 in bEnd.3 cells subjected to OGD.
  
  
  CONCLUSIONS
  These findings suggest that YQFM ameliorates the OGD-induced brain microvascular endothelial cell barrier disruption associated with the NF-κB/p65 and ROCK1/MLC signaling pathways. These data provide new insights into the use of this preparation for treating cerebrovascular diseases.
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China. Electronic address: caoguosheng2006@163.com.
authors.forename: Guo-Sheng
authors.initials: GS
authors.lastname: Cao
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China. Electronic address: chen073011@126.com.
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Chen
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China. Electronic address: cpuzhang27@163.com.
authors.forename: Yuan-Yuan
authors.initials: YY
authors.lastname: Zhang
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China. Electronic address: lifang19870801@126.com.
authors.forename: Fang
authors.initials: F
authors.lastname: Li
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China. Electronic address: 742794892@qq.com.
authors.forename: Chun-Hua
authors.initials: CH
authors.lastname: Liu
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China. Electronic address: 786439572@qq.com.
authors.forename: Xiang
authors.initials: X
authors.lastname: Xiang
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China. Electronic address: yaoyuelingxing@163.com.
authors.forename: Jin
authors.initials: J
authors.lastname: Qi
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China. Electronic address: chaichengzhi@126.com.
authors.forename: Cheng-Zhi
authors.initials: CZ
authors.lastname: Chai
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China. Electronic address: junpingkou@cpu.edu.cn.
authors.forename: Jun-Ping
authors.initials: JP
authors.lastname: Kou
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China. Electronic address: boyangyu59@163.com.
authors.forename: Bo-Yang
authors.initials: BY
authors.lastname: Yu
mesh_terms: D000818:Animals; D001921:Brain / Q000098:blood supply / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002561:Cerebrovascular Disorders / Q000188:drug therapy* / Q000378:metabolism; D051546:Cyclooxygenase 2 / Q000378:metabolism; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D008516:Medicine, Chinese Traditional / Q000379:methods; D051379:Mice; D055806:Microvessels / Q000187:drug effects* / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D011208:Powders / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D051996:Transcription Factor RelA / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D054460:rho-Associated Kinases / Q000378:metabolism*
chemical_list: D004365:Drugs, Chinese Herbal; D053583:Interleukin-1beta; D015850:Interleukin-6; D016328:NF-kappa B; D011208:Powders; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; C584693:yi-qi-fu-mai; D018799:Intercellular Adhesion Molecule-1; D009569:Nitric Oxide; D051546:Cyclooxygenase 2; C515854:Rock1 protein, mouse; D054460:rho-Associated Kinases; D005947:Glucose; D010100:Oxygen
keywords: Ginsenoside Rb1 (PubChem CID: 71317070); Ginsenoside Rc (PubChem CID: 100018); Ginsenoside Re (PubChem CID: 73149); Ginsenoside Rf (PubChem CID: 441922); Ginsenoside Rg 6 (PubChem CID: 91895489); Ginsenoside Rg3 (PubChem CID: 9918693); Microvascular endothelial barrier; NF-κB; Oxygen-glucose deprivation; ROCK1; Schizandrin B (PubChem CID: 108130).; Schizandrol A (PubChem CID: 23915); YiQiFuMai Powder Injection (YQFM)