二甲基亚砜对培养人视网膜色素上皮细胞增殖的影响

Effect of Dimethyl sulfoxide on proliferation of human cultured retinal pigment epithelial cells

目的 探讨细胞分化诱导剂及端粒酶抑制剂二甲基亚砜 (dimethylsulfoxide ,DMSO)对人视网膜色素上皮细胞增殖的抑制作用。方法 本研究分实验组和对照组 ,对照组弃原培养液 ,加无血清培养液继续培养 ,实验组用不同浓度的DMSO 5、10、2 0、4 0mL·L-1分别作用于人RPE细胞 2 4、4 8、72、96h ,用MTT法检测DMSO对hRPE细胞生长的影响 ,流式细胞仪检测DMSO对hRPE细胞周期的影响。结果 10mL·L-1DMSO处理的hRPE细胞从作用 4 8h开始 ,OD值 ( 0 .5 37± 0 .0 10 )小于对照组 ( 0 .6 4 7± 0 .0 2 2 ) ,有显著性差异 (P <0 .0 0 1) ,细胞生长受到抑制 ,且随浓度增加及作用时间延长 ,抑制越显著。流式细胞仪检测 2 0mL·L-1DMSO作用 72h细胞周期发生明显变化 ,G1期细胞由4 1 31%增加至 5 9.6 5 % ,S期由 32 .0 9%减至 2 1.2 1% ,G2 期由 2 6 .6 5 %减至 19 14 % ,细胞生长被阻滞在G1期 ,生长受到抑制。结论 DMSO能阻滞hRPE细胞于G1期 ,抑制hRPE细胞增殖且呈剂量依赖性。

Objective To investigate the inhibiting effect of Dimethyl sulfoxide(DMSO) on the proliferation of cultured human retinal pigment epithelial (hRPE) cells.Methods It was divided into experimental group and control group in this test.The control group,deserted DMEM and added DMEM without containing fetal bovine serum,was continued researching.The cultured cells of the experimental group were treated with DMSO at varying concentrations of 5,10,20,40mL·L -1 , for 24,48,72,96 hours respectively. MTT assay was used to detect inhibiting rates of cultured hRPE cells.Flow cytometry was used to analyze cell cycles.Results Treated with 10 mL·L -1 DMSO for 48 hours,the value of OD(0.537±0.010) began to reduce compared with that of the control group(0.647±0.022),the difference was significant( P <0.001),and the proliferation of cultured cells was inhibited in a dose and time dependent manner .Flow cytometry detected that cell cycles varied significantly treated with 20 mL·L -1 DMSO for 72 hours. The percent of the cultured cells on G 1 phase was increased from 41.31% to 59.65%,the percent on S and G 2 phase were decreased from 32.09% to 21.21% and from 26.65% to 19.14% respectively( P <0 001),hRPE cells were arrested on G 1 phase.Conclusion This study demonstrated that DMSO can arrest RPE cells on G 1 phase and inhibit proliferation of them in a dose dependent manner.