TY - JOUR
T1 - Interleukin-1β and barrier function of retinal pigment epithelial cells (ARPE-19)
T2 - Aberrant expression of junctional complex molecules
AU - Abe, Toshiaki
AU - Sugano, Eriko
AU - Saigo, Yoko
AU - Tamai, Makoto
PY - 2003/9/1
Y1 - 2003/9/1
N2 - PURPOSE. To examine the effects of interleukin (IL)-1β on the resistance and permeability of the tight junctions of cultured retinal pigment epithelial cells. METHODS. A human RPE cell line (ARPE-19) cultured on microporous filter-supports was used. IL-1β and monoclonal anti-IL-1β antibody-treated IL-1β (mAbIL-1β) were added to the standard culture medium. Transepithelial resistance (TER) of confluent RPE cells was measured by epithelial voltmeter. The permeability of the RPE cells to sodium fluorescein, horseradish peroxidase, and inulin was measured. The expression of the occludin and claudin was determined by real-time polymerase chain reaction (PCR), immunohistochemistry, and Western blot analysis. RESULTS. A significantly greater decrease of TER occurred in IL-1β-supplemented medium than in standard medium plus mAbIL-1β after several days of stimulation. A significantly greater increase of sodium fluorescein, horseradish peroxidase, and inulin permeability occurred in IL-1β-supplemented medium than in standard medium. The expression of the occludin gene and some types of claudin genes was observed. The expression of occludin was downregulated and that of claudin-1 upregulated more in IL-1β-supplemented medium than in standard medium by real-time PCR, immunohistochemistry, and Western blot analysis. CONCLUSIONS. The tight junctions of ARPE-19 cells are altered by IL-1β supplementation either directly or through other factors activated by IL-1β. The downregulation of occludin and upregulation of claudin-1 may have participated in the dysfunction of the RPE tight junctions in these in vitro experiments.
AB - PURPOSE. To examine the effects of interleukin (IL)-1β on the resistance and permeability of the tight junctions of cultured retinal pigment epithelial cells. METHODS. A human RPE cell line (ARPE-19) cultured on microporous filter-supports was used. IL-1β and monoclonal anti-IL-1β antibody-treated IL-1β (mAbIL-1β) were added to the standard culture medium. Transepithelial resistance (TER) of confluent RPE cells was measured by epithelial voltmeter. The permeability of the RPE cells to sodium fluorescein, horseradish peroxidase, and inulin was measured. The expression of the occludin and claudin was determined by real-time polymerase chain reaction (PCR), immunohistochemistry, and Western blot analysis. RESULTS. A significantly greater decrease of TER occurred in IL-1β-supplemented medium than in standard medium plus mAbIL-1β after several days of stimulation. A significantly greater increase of sodium fluorescein, horseradish peroxidase, and inulin permeability occurred in IL-1β-supplemented medium than in standard medium. The expression of the occludin gene and some types of claudin genes was observed. The expression of occludin was downregulated and that of claudin-1 upregulated more in IL-1β-supplemented medium than in standard medium by real-time PCR, immunohistochemistry, and Western blot analysis. CONCLUSIONS. The tight junctions of ARPE-19 cells are altered by IL-1β supplementation either directly or through other factors activated by IL-1β. The downregulation of occludin and upregulation of claudin-1 may have participated in the dysfunction of the RPE tight junctions in these in vitro experiments.
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U2 - 10.1167/iovs.02-0867
DO - 10.1167/iovs.02-0867
M3 - Article
C2 - 12939333
AN - SCOPUS:0042283930
VL - 44
SP - 4097
EP - 4104
JO - Investigative Ophthalmology
JF - Investigative Ophthalmology
SN - 0146-0404
IS - 9
ER -