title: Amlodipine suppresses Ang-II-induced endothelium dysfunction by diminishing ROCK1 expression.
abstract: OBJECTIVE
  To investigate the effects and mechanisms of amlodipine therapy on endothelium dysfunction induced by angiotensin-II (Ang-II) stimulation.
  
  
  METHODS
  Human umbilical vein endothelial cells (HUVECs) were used and divided into five groups: Blank control, Ang-II (10(-6 )mol/L), levorotatory amlodipine (5 × 10(-6 )mol/L) + Ang-II (10(-6 )mol/L), dextrorotatory amlodipine (5 × 10(-6 )mol/L) + Ang-II (10(-6 )mol/L) and racemic amlodipine (5 × 10(-6 )mol/L) + Ang-II (10(-6 )mol/L) groups. Twenty-four hours later, HUVECs were collected for evaluating endothelial nitric oxide synthase (eNOS), p-eNOS, rho-associated kinase 1 (ROCK1), Bcl-2 and Bax expressions. Nitric oxide (NO) concentration within endothelium was also detected. Flow cytometry was conducted to assess HUVECs apoptosis.
  
  
  RESULTS
  With 24 hours of Ang-II stimulation, compared to blank control group, expressions of eNOS and p-eNOS and NO production were significantly reduced in Ang-II group (p < 0.05), while adding amlodipine-protected HUVECs from dysfunction induced by Ang-II. In contrast, ROCK1 expression was promoted in Ang-II group (p < 0.05). However, the expression of ROCK1 in each enantiomer of amlodipine group was significantly decreased (p < 0.05). Compared to levorotatory amlodipine group, the magnitude of ROCK1 diminishment in dextrorotatory amlodipine group was more profound (p < 0.05). The pro-survival protein (Bcl-2) was significantly upregulated, while the pro-apoptotic protein (Bax) was significantly downregulated in three amlodipine groups compared to Ang-II group. Flow cytometry revealed that amlodipine therapy could protect HUVECs from apoptosis, and no significant difference between three amlodipine groups was observed.
  
  
  CONCLUSIONS
  Amlodipine could suppress Ang-II-induced endothelial dysfunction and apoptosis through diminishing ROCK1 expression.
authors.affiliation: a Department of Cardiology , The First Affiliated Hospital of Sun Yat-sen University , Guangzhou , China and.
authors.forename: Rulin
authors.initials: R
authors.lastname: Xu
authors.affiliation: b Department of Cardiology , Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of Coronary Heart Disease Prevention, Guangdong General Hospital, Guangdong Academy of Medical Sciences , Guangzhou , China.
authors.forename: Anping
authors.initials: A
authors.lastname: Cai
authors.affiliation: a Department of Cardiology , The First Affiliated Hospital of Sun Yat-sen University , Guangzhou , China and.
authors.forename: Dongdan
authors.initials: D
authors.lastname: Zheng
authors.affiliation: a Department of Cardiology , The First Affiliated Hospital of Sun Yat-sen University , Guangzhou , China and.
authors.forename: Ruofeng
authors.initials: R
authors.lastname: Qiu
authors.affiliation: b Department of Cardiology , Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of Coronary Heart Disease Prevention, Guangdong General Hospital, Guangdong Academy of Medical Sciences , Guangzhou , China.
authors.forename: Liwen
authors.initials: L
authors.lastname: Li
authors.affiliation: b Department of Cardiology , Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of Coronary Heart Disease Prevention, Guangdong General Hospital, Guangdong Academy of Medical Sciences , Guangzhou , China.
authors.forename: Yingling
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: b Department of Cardiology , Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of Coronary Heart Disease Prevention, Guangdong General Hospital, Guangdong Academy of Medical Sciences , Guangzhou , China.
authors.forename: Yingqing
authors.initials: Y
authors.lastname: Feng
authors.affiliation: a Department of Cardiology , The First Affiliated Hospital of Sun Yat-sen University , Guangzhou , China and.
authors.forename: Weiyi
authors.initials: W
authors.lastname: Mai
mesh_terms: D017311:Amlodipine / Q000494:pharmacology*; D000804:Angiotensin II / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D006973:Hypertension / Q000378:metabolism / Q000503:physiopathology*; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000187:drug effects* / Q000378:metabolism; D010750:Phosphoproteins / Q000187:drug effects / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000187:drug effects / Q000378:metabolism; D014662:Vasoconstrictor Agents / Q000494:pharmacology; D051028:bcl-2-Associated X Protein / Q000187:drug effects / Q000378:metabolism; D054460:rho-Associated Kinases / Q000187:drug effects* / Q000378:metabolism
chemical_list: C494826:BAX protein, human; C000595707:BCL2 protein, human; D002121:Calcium Channel Blockers; D010750:Phosphoproteins; D019253:Proto-Oncogene Proteins c-bcl-2; D014662:Vasoconstrictor Agents; D051028:bcl-2-Associated X Protein; D000804:Angiotensin II; D017311:Amlodipine; D009569:Nitric Oxide; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; C515851:ROCK1 protein, human; D054460:rho-Associated Kinases
keywords: Rho-associated kinase; amlodipine; endothelial function