He Hong, Ding Hui, Liao Aiping, Liu Qiong, Yang Jun, Zhong Xingwu
Zhongshan Ophthalmic Center and State Key Laboratory of Ophthalmology, Sun Yat-sen University, Guangzhou, China.
Mol Vis. 2010 Oct 1;16:1913-9.
To investigate the effects of mycophenolate mofetil (MMF) on proliferation and mucin-5AC (MUC5AC) mRNA expression of normal human conjunctival goblet cells (CGCs) in vitro and to understand mechanisms of MMF in treatment of dry eye syndrome at molecular level.
Purified human CGCs were treated with a series of graded concentrations of MMF after being confirmed by immunocytochemistry and flow cytometry. Proliferation and MUC5AC mRNA expression of CGCs were measured by Cell Count Kit-8 (CCK-8) and quantitative nested real-time reverse transcription polymerase chain reaction (QNRT-PCR at 24 h after treatment. The cell proliferation and MUC5AC mRNA expressiion were compared among different doses of MMF.
MMF induced a dose-dependent upregulation of MUC5AC mRNA expression (F=238.851, p<0.01) but a biphase effect on proliferation of the CGCs over 24 h of co-incubation. This biphase effect manifested as a dose-dependent increase in cell numbers with MMF from 0.25 to 2.5 ng/ml, an unchanged population of the cells from 2.5 to 10 ng/ml and a reduced population of the cells from 25 to 100 ng/ml.
MMF exerts biphase effects on cell regeneration and upregulates MUC5AC mRNA expression in CGCs in vitro. It appears that the use of MMF at low concentrations is attractive in dry eye (DE) treatment.
研究霉酚酸酯（MMF）对正常人结膜杯状细胞（CGCs）增殖及黏蛋白-5AC（MUC5AC）mRNA表达的影响，从分子水平了解MMF治疗干眼综合征的机制。
经免疫细胞化学和流式细胞术鉴定后的纯化人CGCs，用一系列梯度浓度的MMF进行处理。采用细胞计数试剂盒-8（CCK-8）和定量巢式实时逆转录聚合酶链反应（QNRT-PCR）在处理24小时后检测CGCs的增殖及MUC5AC mRNA表达。比较不同剂量MMF组的细胞增殖及MUC5AC mRNA表达情况。
MMF诱导MUC5AC mRNA表达呈剂量依赖性上调（F=238.851，p<0.01），但在共孵育24小时期间对CGCs增殖有双相效应。这种双相效应表现为：MMF浓度从0.25至2.5 ng/ml时，细胞数量呈剂量依赖性增加；MMF浓度从2.5至10 ng/ml时，细胞数量不变；MMF浓度从25至100 ng/ml时，细胞数量减少。
MMF对体外CGCs的细胞再生具有双相效应，并上调其MUC5AC mRNA表达。低浓度MMF用于干眼（DE）治疗似乎具有吸引力。
