PMID- 27162720
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20160510
LR  - 20200930
IS  - 2222-3959 (Print)
IS  - 2227-4898 (Electronic)
IS  - 2222-3959 (Linking)
VI  - 9
IP  - 4
DP  - 2016
TI  - Cytotoxicity of pilocarpine to human corneal stromal cells and its underlying 
      cytotoxic mechanisms.
PG  - 505-11
LID - 10.18240/ijo.2016.04.05 [doi]
AB  - AIM: To examine the cytotoxic effect of pilocarpine, an anti-glaucoma drug, on 
      human corneal stromal (HCS) cells and its underlying cytotoxic mechanisms using 
      an in vitro model of non-transfected HCS cells. METHODS: After HCS cells were 
      treated with pilocarpine at a concentration from 0.15625 g/L to 20.0 g/L, their 
      morphology and viability were detected by light microscopy and MTT assay. The 
      membrane permeability, DNA fragmentation and ultrastructure were examined by 
      acridine orange (AO)/ethidium bromide (EB) double-staining. DNA electrophoresis 
      and transmission electron microscopy (TEM), cell cycle, phosphatidylserine (PS) 
      orientation and mitochondrial transmembrane potential (MTP) were assayed by flow 
      cytometry (FCM). And the activation of caspases was checked by ELISA. RESULTS: 
      Morphology observations and viability assay showed that pilocarpine at 
      concentrations above 0.625 g/L induced dose- and time-dependent morphological 
      abnormality and viability decline of HCS cells. AO/EB double-staining, DNA 
      electrophoresis and TEM noted that pilocarpine at concentrations above 0.625 g/L 
      induced dose- and/or time-dependent membrane permeability elevation, DNA 
      fragmentation, and apoptotic body formation of the cells. Moreover, FCM and ELISA 
      assays revealed that 2.5 g/L pilocarpine also induced S phase arrest, PS 
      externalization, MTP disruption, and caspase-8, -9 and -3 activation of the 
      cells. CONCLUSION: Pilocarpine at concentrations above 0.625 g/L (1/32 of its 
      clinical therapeutic dosage) has a dose- and time-dependent cytotoxicity to HCS 
      cells by inducing apoptosis in these cells, which is most probably regulated by a 
      death receptor-mediated mitochondrion-dependent signaling pathway.
FAU - Yuan, Xiao-Long
AU  - Yuan XL
AD  - Laboratory for Corneal Tissue Engineering, College of Marine Life Sciences, Ocean 
      University of China, Qingdao 266003, Shandong Province, China.
FAU - Wen, Qian
AU  - Wen Q
AD  - Laboratory for Corneal Tissue Engineering, College of Marine Life Sciences, Ocean 
      University of China, Qingdao 266003, Shandong Province, China.
FAU - Zhang, Meng-Yu
AU  - Zhang MY
AD  - Laboratory for Corneal Tissue Engineering, College of Marine Life Sciences, Ocean 
      University of China, Qingdao 266003, Shandong Province, China.
FAU - Fan, Ting-Jun
AU  - Fan TJ
AD  - Laboratory for Corneal Tissue Engineering, College of Marine Life Sciences, Ocean 
      University of China, Qingdao 266003, Shandong Province, China.
LA  - eng
PT  - Journal Article
DEP - 20160418
PL  - China
TA  - Int J Ophthalmol
JT  - International journal of ophthalmology
JID - 101553860
PMC - PMC4853343
OTO - NOTNLM
OT  - apoptosis
OT  - cytotoxicity
OT  - human corneal stromal cells
OT  - mitochondrion
OT  - pilocarpine
EDAT- 2016/05/11 06:00
MHDA- 2016/05/11 06:01
PMCR- 2016/04/18
CRDT- 2016/05/11 06:00
PHST- 2015/03/18 00:00 [received]
PHST- 2015/09/29 00:00 [accepted]
PHST- 2016/05/11 06:00 [entrez]
PHST- 2016/05/11 06:00 [pubmed]
PHST- 2016/05/11 06:01 [medline]
PHST- 2016/04/18 00:00 [pmc-release]
AID - ijo-09-04-505 [pii]
AID - 10.18240/ijo.2016.04.05 [doi]
PST - epublish
SO  - Int J Ophthalmol. 2016 Apr 18;9(4):505-11. doi: 10.18240/ijo.2016.04.05. 
      eCollection 2016.