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Junko Hirata, Ji-Ae Ko, Hideki Mochizuki, Kunihiko Funaishi, Ken Yamane, Yoshiaki Kiuchi; Oxidative stress affects tight junction expression in Human retinal pigment epithelium. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1801.
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In age-related macular degeneration (AMD), it has been previously shown that cell adhesion molecules play a critical role in the retinal pigment epithelium. We have now examined the effects of oxidative stress on the expression of junctional proteins in RPE cells.
A concentration of 5×105 Human retinal pigment epithelium (ARPE-19) cells were cultured and serum starved for 24 hours. And, the cells were treated with 500μM hydrogen peroxide (H2O2) for indicated times. The expression of junctional proteins (claudin-1, ZO-1 and N-cadherin) were examined by RT-PCR, immunoblot, and immunofluorescence analyses.
RT-PCR analysis revealed that the abundance of claudin-1 was increased in 8, 10 hours after H2O2 treatment. Immunoblot analysis also showed that the amounts of claudin-1 in ARPE-19 were increased in 8, 10 hours after H2O2 treatment. But, The increases in the amounts of these mRNAs and proteins were maximal at 10 and were decreased gradually, according to time course. But other junctional proteins, such as ZO-1 and N-cadherin, didn’t any changed.
These results suggest that claudin-1 may play an important role in intercellular communication between RPE cells. In brief, it is possible that clauidn-1 is important key factor in early steps of AMD.
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