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PMID- 28418499
OWN - NLM
STAT- In-Process
DA  - 20170418
LR  - 20170418
IS  - 1552-5783 (Electronic)
IS  - 0146-0404 (Linking)
VI  - 58
IP  - 4
DP  - 2017 Apr 01
TI  - Sphingosine-1-Phosphate (S1P)-Related Response of Human Conjunctival Fibroblasts 
      After Filtration Surgery for Glaucoma.
PG  - 2258-2265
LID - 10.1167/iovs.16-21288 [doi]
AB  - Purpose: To investigate levels of sphingosine-1-phosphate (S1P) in aqueous fluid 
      samples taken before and after filtration surgery and S1P-induced human
      conjunctival fibroblast (HCF) responses. Methods: Levels of S1P and its related
      sphingophospholipids in aqueous fluid obtained immediately before and after
      filtration surgery were determined by liquid chromatography-tandem mass
      spectrometry. HCFs were used for all in vitro experiments. The expression of five
      S1P receptor subtypes in HCFs was examined by quantitative real-time PCR. The
      effect of S1P and receptor-specific antagonists on HCF viability and cell
      migration was assessed by WST-1 assay and scratch migration assay, respectively. 
      Differentiation to myofibroblasts and extracellular matrix production was
      evaluated by examining changes in F-actin, alpha-smooth muscle actin (alphaSMA), 
      and collagen expression with immunocytochemistry, Western blotting, and collagen 
      accumulation assay, respectively. Results: No significant S1P levels in the
      aqueous fluid samples were detectable immediately before surgery, but
      postoperative levels of several lysophospholipids, including S1P, dehydro-S1P,
      and sphingosine, were significantly increased to bioactive concentrations in
      aqueous fluid in the blebs (P < 0.0001). mRNA expression of the three main S1P
      receptor subtypes was detected in HCFs. Although S1P levels did not influence HCF
      proliferation, S1P enhanced cell migration, which could be inhibited by the S1P2 
      antagonist JTE 013. F-actin, alphaSMA, and collagen expression was significantly 
      increased by S1P stimulation and was reduced by JTE 013. Conclusions: Bioactive
      S1P concentrations were present in the aqueous fluid at the end of filtration
      surgery. S1P activated HCFs via S1P2 receptors. These results revealed the
      potential of S1P2 antagonists in preventing scarring after glaucoma filtration
      surgery.
FAU - Aoyama-Araki, Yuka
AU  - Aoyama-Araki Y
AD  - Department of Ophthalmology, The University of Tokyo, Tokyo, Japan.
FAU - Honjo, Megumi
AU  - Honjo M
AD  - Department of Ophthalmology, The University of Tokyo, Tokyo, Japan.
FAU - Uchida, Takatoshi
AU  - Uchida T
AD  - Department of Ophthalmology, The University of Tokyo, Tokyo, Japan 2Senju
      Pharmaceutical Co., Ltd., Kobe, Japan.
FAU - Yamagishi, Reiko
AU  - Yamagishi R
AD  - Department of Ophthalmology, The University of Tokyo, Tokyo, Japan.
FAU - Kano, Kuniyuki
AU  - Kano K
AD  - Department of Molecular and Cellular Biochemistry, Tohoku University, Sendai,
      Japan.
FAU - Aoki, Junken
AU  - Aoki J
AD  - Department of Molecular and Cellular Biochemistry, Tohoku University, Sendai,
      Japan.
FAU - Aihara, Makoto
AU  - Aihara M
AD  - Department of Ophthalmology, The University of Tokyo, Tokyo, Japan.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
EDAT- 2017/04/19 06:00
MHDA- 2017/04/19 06:00
CRDT- 2017/04/19 06:00
AID - 2621648 [pii]
AID - 10.1167/iovs.16-21288 [doi]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 2017 Apr 1;58(4):2258-2265. doi:
      10.1167/iovs.16-21288.

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