ObjectiveTo observe the effect of triamcinolone acetonide(TA) on activation and barrier function of human retinal pigment epithelium （RPE）.MethodsARPE-19 cells were cultured in 96well tissue culture plate. Four weeks later, TA with different concentration (0.02 and 0.05 mg/ml)was added to the cells and culture for 3 or 7 days. The activation of ARPE-19 cells was assessed by methyl thiazolyl tetrazolium (MTT). ARPE-19 cells were cultured on polyester microporous filters for 4 weeks, and the transepithelial resistance (TER) was recorded. TA (0.02 and 0.05 mg/ml) was added to the culture fluid respectively, and after cultured for 1 week TER was measured again. The RPE permeability was detected by enzymelinked immunosorbent assay (ELISA) with horse radish peroxidase as the tracer. ResultsIn the culture fluid with 002 mg/ml TA cultured for 3 or 7 days, the average survival rate of ARPE-19 cells was 93.70% and 90.63% respectively, without statistic difference compared with the control (P=0.147, 0.091). While in the 0.05 mg/ml TA group after cultured for the same duration, the activation of ARPE-19 cells decreased significantly compared with the control (with the average survival rate of 87.75% and 88.98%; P=0.025, 0.043). One week after cultured with TA, TER decreased significantly while permeability improved obviously in the 2 TA groups compared to the control (Plt;0.001; 0.001lt;Plt;0.05).ConclusionTA may decrease the activation of and destroy the barrier function of ARPE-19 cells. (Chin J Ocul Fundus Dis, 2005,21:237-239)
Objective To investigate the effects of QUE on proliferation and DNA synthesis of cultured retinal pigment epithelium(RPE) cells with or without EGF. Methods With or without EGF, cultured RPE cells were treated with QUE by various concentrations(200,100,50,1mu;mol/L) and with QUE 200mu;mol/L at different times(24-168 hr), cells proliferation and DNA synthesis were evaluated by cell count method and the uptake of thymidine. The viability of cells was determined by trypanblue exclusion. Results The best concentration of QUE which inhibits proliferation and DNA synthesis of PRE cells was 200mu;mol/L. The significant inhibition effect of QUE occurred at 48hr, and the best inhibition of QUE occurred at 96hr. QUE had more powerful effect of antiproliferation on RPE cells, and the viability of RPE cells was over85%. Conclusion The results suggested that QUE could inhibit the proliferation of RPE cells in a dose-dependent and time-dependent manner, especially inhibit the proliferation induced by EGF stimulating. QUE had no cyto-toxic effect on RPE cells cultured in vitro. (Chin J Ocul Fundus Dis,1999,15:27-29)
OBJECTIVE:To observe the effect of dexamethasone to intracellular free Ca2+ of frozen RPE cells. METHODS:The cultured human RPE cells were frozen for 30s at --70deg;C. The RPE cells were loaded with Fura-2/AM and analyzed using a digital imaging microscopy system,the effect of dexamethasone to intracellular free Ca2+ was measured at a serial concentration of 40, 60,100,150,200mu;g/ml. RESULTS:The concentration of intracellular free Ca in frozen human RPE cells was increased to 18.6%~29.8% by dexamethasone at concenlration of 40mu;g/ml~60mu;g/ml,while was decreased to 28.4%~35.2% at 150mu;g/ml~200mu;g/ml. CONCLUSIONS:Effect of dexamethasone showed two aspects of effect to frozen cultured human RPE ceils,that it was inhibitor at high concentration and stimulator at low concentration （Chin J Ocul Fundus Dis,1997,13: 86-88）