title: Rho-Kinase inhibitor fasudil suppresses high glucose-induced H9c2 cell apoptosis through activation of autophagy.
abstract: BACKGROUND
  Cardiac cell apoptosis plays a crucial role in the progression of diabetic cardiomyopathy. Recent studies have shown that fasudil, a Rho-kinase (ROCK) inhibitor, inhibits cardiac cell apoptosis; however, the underlying mechanism remains unclear.
  
  
  OBJECTIVE
  This study aimed to investigate whether fasudil protects H9c2 cells from high glucose-induced apoptosis via activation of autophagy.
  
  
  METHODS
  Rat cardiomyocyte H9c2 cells were treated with high glucose and used as a diabetic cardiomyopathy model. Cell survival rate, apoptosis, and subcellular morphology were examined using the MTT assay, flow cytometry, and electron microscopy, respectively. ROCK1 and ROCK2 mRNA levels were determined using quantitative real-time PCR. Bcl-2 and Bax, myosin phosphatase target subunit-1 (MYPT-1), phosphorylated (p)-MYPT1, LC3-II/LC3-I, Beclin-1, soluble and insoluble P62 protein levels were determined by Western blot analysis.
  
  
  RESULTS
  Fasudil reversed the high glucose-induced inhibition of cell proliferation and suppressed high glucose-induced early apoptosis. Fasudil also reversed the high glucose-suppressed Bcl-2 levels and decreased the high glucose-induced Bax levels. Further, Fasudil suppressed ROCK levels, expression, promoted autophagy via increasing the LC3-II/LC3-I ratio, Beclin-1 expression, and the number of autophagosomes in H9c2 cells treated with high glucose. These effects of fasudil were abrogated by 3-methyladenine (3-MA), an autophagy inhibitor.
  
  
  CONCLUSIONS
  Fasudil inhibited high glucose-induced apoptosis in rat H9c2 cells through activating autophagy.
authors.affiliation: Department of Cardiology, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
authors.forename: Huikuan
authors.initials: H
authors.lastname: Gao
authors.affiliation: Department of Infection, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Hou
authors.affiliation: Department of Cardiology, Hangzhou First People's Hospital, Zhejiang, China.
authors.forename: Ruiqing
authors.initials: R
authors.lastname: Dong
authors.affiliation: Department of Cardiology, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
authors.forename: Zefeng
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Cardiology, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
authors.forename: Can
authors.initials: C
authors.lastname: Zhao
authors.affiliation: Department of Cardiology, Baotou Central Hospital, Inner Mongolia, China.
authors.forename: Wurina
authors.initials: W
authors.lastname: Tang
authors.affiliation: Department of Cardiology, Beijing Friendship Hospital, Capital Medical University, Beijing, China. wuyongquan67@163.com.
authors.forename: Yongquan
authors.initials: Y
authors.lastname: Wu
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D058065:Diabetic Cardiomyopathies / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D005947:Glucose / Q000633:toxicity*; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000201:enzymology / Q000648:ultrastructure; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; D047428:Protein Kinase Inhibitors; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C578418:ROCK1 protein, rat; C580818:ROCK2 protein, rat; D054460:rho-Associated Kinases; D005947:Glucose; C049347:fasudil
keywords: Autophagy; Cardiac apoptosis; Diabetic cardiovascular disease; Fasudil; Rho kinase