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| The effects of indomethacin on proliferation and DNA synthesis and apoptosis in cultured human fetal retinal pigment epithelium cells in vitro |
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| 作者：LI-Wensh… 文章来源：Eye Hospital of Wenzhou Medical College, Wenzhou 325027, China 点击数1250 更新时间：2006/6/23 15:28:38 文章录入：李文生 责任编辑：毛进 |
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| [Absract] Objective: To investigate the inhibitive effects indomethacin (IN) on proliferation and DNA synthesis and apoptosis of cultured human fetal retinal pigment epithelium(hRPE) cells in vitro. Methods: Cultured hRPE cells were treated with IN by various concentrations 100, 200, 400, 600, 800, 1000μmol•L-1for 12h. After 12h, cells proliferation were measured with methyl thiazolyl tetrazolium (MTT) assay method and 50, 100, 200, 400，600μmol•L-1 IN for 24h. After 24h, the amount of DNA was determined by the absorbance at 280 nm of Nucleic Acid δProtein Analysis. Cultured RPE cells were treated with IN by various concentrations 100, 200, 400, 600μmol•L-1for 24h and 600μmol/L IN for 6h, 12h, 24h, the quantitative and the qualitative analysis of apoptotic RPE cells were assessed using an acridine orange (AO) staining method and transmission electron microscopy (TEM) respectively. Results: The A values of 100, 200, 400, 600,800,1000μmol•L-1 IN were (0.2367±0.0546), (0.1687±0.0695), (0.0819±0.03461), (0.0656±0.01759), (0.0554±0.02865), (0.0508±0.02775) compared to (0.2674±0.04302) of A value of 0μg•L-1 IN respectively, t value was 1.44，4.621，8.682，9.447，9.925，10.140 and P values were 0.158,0.000，0.000，0.000，0.000，0.000 (t-test). The DNA concentrations of 50, 100, 200, 400，600μmol•L-1 IN were (101.1712±15.5124), (88.6400±13.5845), (72.3651±7.7969), (59.9089±10.7229), (51.2236±8.7757)μg/ml compared to (213.7351±83.1572)μg• ml-1 of DNA concentration of 0μg•L-1 IN respectively, t values were 5.481，6.091，6.883，7.490，7.912， P values were 0.000，0.000，0.000，0.000，0.000 (t-test). AO staininng in apoptotic RPE cells showed typical apoptosis with chromation condensation and reduction of nuclear size and cell volume and orange-stained condensed nuclei.The RPE apoptotic numbers of 200μmol/L,400μmol/L, 600μmol/L IN was (2.7000±1.3375), (5.1000±1.3703), (6.8000±1.6865) compared to (0.2000±0.4216) of number of 0μmol/L IN respectively, t value was 4.624，9.062，12.206 and P value was 0.000，0.000， 0.000 (t-test) after 72h; The RPE apoptotic numbers of 12h, 24h was (4.4000±0.8433), (6.4000±1.2649) compared to (0.2000±0.4216) of number of 12h respectively, t value was 8.617, 13.274 and P value was 0.000，0.000 (t-test) by 600μmol•L-1IN , the more concentration of IN, the more apoptosis of RPE cells. Conclusion: The data suggested that IN could inhibit the proliferation and DNA synthesis and apoptosis in cultured of hRPE cells in vitro in a dose and course-dependent manner. |
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