PMID- 28002844
OWN - NLM
STAT- MEDLINE
DCOM- 20170615
LR  - 20171220
IS  - 1552-5783 (Electronic)
IS  - 0146-0404 (Linking)
VI  - 57
IP  - 15
DP  - 2016 Dec 1
TI  - Activation of the Rho/Rho Kinase Signaling Pathway Is Involved in Cell Death of 
      Corneal Endothelium.
PG  - 6843-6851
LID - 10.1167/iovs.16-20123 [doi]
AB  - PURPOSE: Rho kinase (ROCK) pathways control fundamental cell functions, making 
      ROCK an important therapeutic target in several pathophysiologic conditions. The 
      purpose of this study was to investigate whether inhibition of ROCK can suppress 
      apoptosis of the corneal endothelium and to determine the role of ROCK signaling 
      in regulating apoptosis. METHODS: The effects of inhibitors of ROCK or myosin 
      light chain (MLC) were evaluated in cultured monkey corneal endothelial cells 
      (MCECs) irradiated with ultraviolet (UV) (100 J/m2) to induce apoptosis. Annexin 
      V and TUNEL staining and Western blot for apoptosis-related proteins and focal 
      adhesion complexes were then performed. RhoA activation was further evaluated by 
      pull-down assays. ROCK inhibitor and caspase inhibitor effects on apoptosis were 
      also evaluated in MCECs treated with ethylene glycol tetraacetic acid (EGTA) to 
      induce MLC phosphorylation. RESULTS: ROCK or MLC inhibition suppressed the 
      caspase-3 cleavage and Annexin V and TUNEL expression typically seen during 
      UV-mediated apoptosis of MCECs. The apoptotic stimulus activated RhoA and then 
      induced phosphorylation of MLC via ROCK activation. EGTA-mediated phosphorylation 
      of MLC was sufficient to induce the loss of cell contact with the substrate and 
      subsequent apoptosis. Western blot showed that ROCK inhibition upregulated the 
      expression of the focal adhesion complex in adhered cells, following UV stress. 
      CONCLUSIONS: Apoptotic stimuli activated Rho/ROCK/MLC phosphorylation in the 
      corneal endothelium, and subsequent actomyosin contraction induced apoptosis by 
      loss of cell adhesion. ROCK inhibition suppressed MLC phosphorylation and 
      subsequent cell death, and it counteracted the loss of cell adhesion by 
      activating the focal adhesion complex.
FAU - Okumura, Naoki
AU  - Okumura N
AD  - Department of Biomedical Engineering, Faculty of Life and Medical Sciences, 
      Doshisha University, Kyotanabe, Japan.
FAU - Fujii, Keita
AU  - Fujii K
AD  - Department of Biomedical Engineering, Faculty of Life and Medical Sciences, 
      Doshisha University, Kyotanabe, Japan.
FAU - Kagami, Takato
AU  - Kagami T
AD  - Department of Biomedical Engineering, Faculty of Life and Medical Sciences, 
      Doshisha University, Kyotanabe, Japan.
FAU - Makiko, Nakahara
AU  - Makiko N
AD  - Department of Biomedical Engineering, Faculty of Life and Medical Sciences, 
      Doshisha University, Kyotanabe, Japan.
FAU - Kitahara, Miu
AU  - Kitahara M
AD  - Department of Biomedical Engineering, Faculty of Life and Medical Sciences, 
      Doshisha University, Kyotanabe, Japan.
FAU - Kinoshita, Shigeru
AU  - Kinoshita S
AD  - Department of Frontier Medical Science and Technology for Ophthalmology, Kyoto 
      Prefectural University of Medicine, Kyoto, Japan.
FAU - Koizumi, Noriko
AU  - Koizumi N
AD  - Department of Biomedical Engineering, Faculty of Life and Medical Sciences, 
      Doshisha University, Kyotanabe, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - EC 2.7.11.1 (rho-Associated Kinases)
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - Cell Death/*physiology
MH  - Cells, Cultured
MH  - Disease Models, Animal
MH  - Endothelium, Corneal/*metabolism/pathology
MH  - Flow Cytometry
MH  - Immunohistochemistry
MH  - In Situ Nick-End Labeling
MH  - Macaca fascicularis
MH  - Phosphorylation
MH  - Rabbits
MH  - Signal Transduction
MH  - rho-Associated Kinases/*metabolism
EDAT- 2016/12/22 06:00
MHDA- 2017/06/16 06:00
CRDT- 2016/12/22 06:00
PHST- 2016/12/22 06:00 [entrez]
PHST- 2016/12/22 06:00 [pubmed]
PHST- 2017/06/16 06:00 [medline]
AID - 2594998 [pii]
AID - 10.1167/iovs.16-20123 [doi]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 2016 Dec 1;57(15):6843-6851. doi: 
      10.1167/iovs.16-20123.