体外培养人角膜上皮细胞中CD44的表达及IFN-γ和雷公藤多甙对其的影响

Expression of CD44 in cultured human corneal epithelium and the influence of IFN-γ and Glucosida Tripterygii TOTA to it

目的探讨黏附分子CD44在角膜上皮细胞中的表达及干扰素-γ(IFN-γ)和雷公藤多甙对角膜上皮细胞中CD44表达的影响。方法采用细胞培养和流式细胞技术,分别观察同一培养时间(24h)不同IFN-γ含量和同一IFN-γ含量不同培养时间对人角膜上皮细胞CD44表达的影响及0.03%雷公藤多甙对角膜上皮细胞中CD44表达的影响。结果体外培养的人角膜上皮细胞可基础表达一定量的CD44(1.944±0.237),IFN-γ可上调CD44的表达,其上调作用呈量和时间依赖性(P<0.05)。雷公藤多甙可以使角膜上皮中CD44的表达降低(P<0.01)。结论IFN-γ可以使角膜上皮细胞中CD44的表达升高,使排斥反应加剧;雷公藤多甙可以抑制角膜上皮细胞中CD44的表达,从而抑制角膜移植排斥反应。

Objective Along with the unceasing investigation of immunological rejection molecular mechanism,researchers found that adhesion molecule has a close relation to organ-graft refection.This experiment observed the expression of CD44 in normal human corneal epithelial cells and evaluated the influence of interferon-γ（IFN-γ） and Glucosida Tripterygii TOTA on expression of CD44.MethodsHuman corneal epithelial cells were isolated from 11 normal corneas of donors and cultured in SHEM culture media.The cultured cells were randomly assigned to 0,100,200,500,1000U/mL IFN-γ for culture for 24 hours respectively（group A,B,C,D,E） and 500U/mL of IFN-γ was used to treat cultured cells for 48 and 72 hours（ group F,G）.Corneal epithelial cells were treated by 500U/mL IFN-γ for 24 hours then followed free-drug culture for 24 hours in group H.0.03%Glucosida Tripterygii TOTA was added to the media for 24 hours after 500U/mL IFN-γ cultured 24 hours in group I.Flow cytometry was used to analyze the fluorescence index（FI） of CD44 in cultured human corneal epithelium cells.ResultsCultured human corneal epithelial cells constitutively expressed certain amount of CD44（1.944±0.237）（group A）,and the FI of CD44 was significantly elevated in different concentrations of IFN-γ groups and different time（P〈0.05） at a concentration-and time-indipendent manner.The FI of CD44 was 3.213±0.073 in Glucosida Tripterygii TOTA group（group I）,and that without Glucosida Tripterygii TOTA group（group H） was 3.618±0.061,showing a significant difference between them（P〈0.01）.ConclusionIFN-γ can enhance the expression of CD44 in human corneal epithelial cells and thereby aggravate the corneal graft rejection.Glucosida Tripterygii TOTA can depress the CD44 expression.