全反式视黄酸对体外培养大鼠Müller细胞生长的影响

目的:观察全反式视黄酸(All-trans retinoic acid,RA)对体外培养大鼠Müller细胞生长的影响。方法:用不同浓度的RA作用于培养的Müller细胞,利用相差显微镜、MTT法、细胞计数法、流式细胞仪检测RA对Müller细胞形态、生长状况及其凋亡的影响。结果:RA在低浓度(<0.1μM)时对Müller细胞的抑制作用不明显。当RA在高浓度(1μM、10μM、100μM)时产生明显抑制作用,细胞形态出现显著变化,48h较24h更明显。细胞计数显示20μM的RA作用48h后引起细胞数目明显减少。流式细胞仪检测结果显示不同浓度的RA(5μM、10μM、20μM)均可诱导Müller细胞凋亡,20μM的RA作用48h时细胞凋亡率最高(P<0.01),凋亡指数为(35.87±7.40)%。结论:RA能抑制Müller细胞的增殖、促进其凋亡,并具有浓度和时间依赖性。

Effects of All-trans Retinoic Acid on the Rat Müller Cell in Vitro

PURPOSE: To investigate the effects of various concentrations of all-trans retinoic acid (RA) on the rat Müller cells in vitro. METHODS: A rat Müller cell line was used in this study. Rat Müller cells were cultured with varying levels of RA for 24 h to 48 h. We examined cellular morphology under phase contrast microscope, cell proliferation using MTT assay, viable cell numbers by hemocytometer counting and cell apoptosis with flow cytometry. RESULTS: At lower concentration (< 0.1 microM), RA didn't change the cell's appearance and didn't inhibit the cell proliferation (P > 0.05). However, at higher concentration (1 miroM, 10 microM, 100 microM), RA could inhibit the proliferation of Müller cells accompanied with morphological changes in a time- and dose-dependent manner. Furthermore, different concentration of RA (5 microM, 10 microM, 20 microM) could induce cell apoptosis. When using 20 microM RA to treat cells for 48 h, a significant decrease in cell numbers and an obvious increment of apoptotic cells were observed. The percentage of apoptotic cell was (35.87 +/- 7.40)% (P < 0.01, vs. control). CONCLUSIONS: RA can inhibit the proliferation and induce the apoptosis of müller cells in a time- and dose-dependent manner.