Synthetic bile acid derivatives induce nonapoptotic death of human retinal pigment epithelial cells.
OBJECTIVE
To study whether the synthetic ursodeoxycholic acid (UDCA) and chenodeoxycholic acid (CDCA) derivatives, which we have synthesized and have reported their apoptosis-inducing effect, have the effect on the proliferation of retinal pigment epithelial cells.


METHODS
UDCA, CDCA, and their synthetic derivatives were administered in culture to the human retinal pigment cell line, ARPE-19. The effect on cell viability and growth was assessed by trypan blue dye exclusion. In order to evaluate the type of cell death, mitochondrial membrane potential assay, DNA electrophoresis, TUNEL assay, nuclear staining and Western blotting for caspase-3 and poly(ADP-ribose) polymerase (PARP) activities were conducted.


RESULTS
Unlike UDCA and CDCA, which did not exhibit a significant effect on viability, their synthetic derivatives decreased the viability of ARPE-19 cells in a concentration-dependent manner. The cells treated with the synthetic derivatives did not demonstrate the characteristic findings of apoptosis, such as DNA ladder, DNA fragmentation, nuclear condensation or fragmentation, and caspase-3 and PARP activation. The reduction of mitochondrial membrane potential was shown. In electron microscopical study nuclear condensation was not shown.


CONCLUSIONS
The synthetic UDCA and CDCA derivatives induced nonapoptotic death of ARPE-19 cells.
Yoon
H S
HS
Department of Ophthalmology, Dong-A University College of Medicine and Institute of Medical Science, Pusan, South Korea.
Rho
J H
JH
Yoo
K W
KW
Park
W C
WC
Rho
S H
SH
Choi
Y H
YH
Suh
H
H
Kim
N D
ND
Yoo
K S
KS
Yoo
Y H
YH
D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects*; D002478:Cells, Cultured; D002635:Chenodeoxycholic Acid / Q000031:analogs & derivatives / Q000494:pharmacology*; D004247:DNA / Q000032:analysis; D053938:DNA Fragmentation; D004305:Dose-Response Relationship, Drug; D006801:Humans; D020287:In Situ Nick-End Labeling; D008564:Membrane Potentials / Q000502:physiology; D008928:Mitochondria / Q000187:drug effects; D010857:Pigment Epithelium of Eye / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D014343:Trypan Blue / Q000378:metabolism; D014580:Ursodeoxycholic Acid / Q000031:analogs & derivatives / Q000494:pharmacology*
D002635:Chenodeoxycholic Acid; D014580:Ursodeoxycholic Acid; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D014343:Trypan Blue