1,25-二羟维生素D_3对体外人晶状体上皮细胞增殖的影响

目的：研究1,25二羟维生素D3[1,25-（OH）2D3,简称D3]对体外培养的人晶状体上皮细胞（human lens epithelial cells,hLEC）增殖的影响。方法：人晶状体上皮细胞系SRA01/04常规培养,取对数期细胞接种于96孔培养板,将含有细胞和培养液不含D3的空白对照组,只含有培养液不含有细胞及D3的调零组,含有无水乙醇（10-4mol/L）、细胞和培养液的阴性对照组,含有不同浓度（10-10,10-9,10-8,10-7mol/L）的D3组,分别处理24,48,72h后,采用MTT法检测各组细胞在490nm波峰处吸光度A值,计算细胞生长抑制率;FCM检测10-7mol/LD3处理细胞48h后细胞周期的改变。结果：24和48h10-8和10-7mol/LD3组A值均较对照组下降（P〈0.01）,72h各处理组和对照组差异无统计学意义。10-7mol/LD348h后,G1和S期细胞数占总细胞数百分比分别为（65.3±2.4）%和（30.0±1.4）%,与对照组[（57.4±1.5）%和（39.0＋1.4）%]相比差异有统计学意义（P〈0.01）。结论：1,25-（OH）2D3在10-7和10-8mol/L对细胞增殖具有抑制作用,使hLEC部分阻滞于细胞周期的G1期。

Effect of 1,25-dihydroxyvitamin D3 on proliferation of human lens epithelial cells in vitro

AIM：To study the effect of 1,25-dihydroxyvitamin D3 ／[1,25-（OH）2D3,D3／] on proliferation of human lens epithelial cells（hLEC） in vitro.·METHODS：hLEC line SRA01/04 in logarithmic phase were inoculated to 96-well plate,and divided into blank group containing cells and culture medium without D3,zero group containing culture medium without cells and D3,negative control group containing absolute alcohol（10-4mol/L）,cells and culture medium,D3 groups containting different concentrations（10-10,10-9,10-8,10-7mol/L）,after treating for 24,48 and 72 hours,MTT assay was used for detecting the values of absorbance A at 490nm in each groups,and inhibiting rate of cell growth was calculated.Cell cycle changes with D3-treated group（10-7mol/L D3） for 48 hours was detected by flow cytometry assay.·RESULTS：The values of absorbance A in D3-treated groups（10-7,10-8mol/L） decreased compared with control group（P〈0.01） at 24,48 time points.There was no significant difference between each group at 72 time point and control group.The cells were treated with D3（10-7mol/L） for 48 hours,and percentage of cells in G1 and S phase was（65.3±2.4）% and（30.0±1.4）%,and there was statistically difference compared to the control group ／[（57.4± 1.5）% and（39.0 ＋ 1.4）%／],（P〈0.01）.·CONCLUSION：The proliferation of hLEC was inhibited by 1,25-dihydroxyvitamin D3（10-7and 10-8mol/L）,and the cells were partially blocked at G1 phase of cell cycle.