双氢青蒿素对体外培养兔角膜上皮细胞毒性作用的实验研究

目的 评价双氢青蒿素对体外培养的兔角膜上皮细胞的毒性作用。设计 实验研究。研究对象 体外培养兔角膜上皮细胞。方法 体外培养兔角膜上皮细胞，加入不同稀释浓度（1.6%、0.8%、0.4%、0.2%、0.1%、0.05%和0.025%）的双氢青蒿素共同孵育，同时以洗必泰和聚六亚甲基双胍（PHMB）做对照药物。在培养后2、6、24小时，光学显微镜下观察细胞形态，LIVE/DEAD染色观察药物作用后兔角膜上皮细胞的活性改变。主要指标 细胞形态改变。结果 培养后2、6小时，0.4%双氢青蒿素作用后的细胞出现圆化，细胞间隙加宽；0.0125%洗必泰作用后的细胞开始出现细胞间隙加宽，边界不清；0.003%PHMB作用后的细胞开始出现细胞边界不清，细胞内颗粒样改变。培养后24小时，0.1%的双氢青蒿素作用的角膜上皮细胞尚存在活性，0.025%的双氢青蒿素对角膜上皮细胞无明显影响。0.006%洗必泰和0.003%PHMB可导致角膜上皮细胞的死亡。结论 双氢青蒿素对体外培养的兔角膜上皮细胞的毒性较低，其对活体角膜细胞的作用尚待进一步研究。

Experimental study on the toxic effect of dihydroartemisinine on rabbit corneal epithelial cells cultured in vitro

Objective To evaluate the toxic effect of dihydroartemisinine on rabbit corneal epithelial cells in vitro. Design Experimental study. Participants Rabbit corneal epithelial cells in vitro.  Methods Different concentrations (1.6%, 0.8%, 0.4%, 0.2%, 0.1%, 0.05%, 0.025%) of dihydroartemisinin was added to the cultured rabbit corneal epithelial cells in vitro. Chlorhexidine and PHMB were  used as controls. The morphology of cells were observed in 2, 6 and 24 hr under optical microscope. LIVE/DEAD staining was used to observe the changes of the rabbit corneal epithelial cells after drug action under fluorescence microscope. Main Outcome Measures Morphological changes of the epithelial cells under optical microscope and fluorescence microscope. Results At the point of 2 and 6 hr after culturing, the rims of cells were rounded and the gaps were widened under the action of 0.4% dihydroartemisinine. The gaps of cells began to appear widening and unclear under the action of 0.0125% chlorhexidine. The boundary of cells began to appear unclear and there were intracellular particle changes under the action of 0.003% PHMB. At the point of 24 hr after culturing, corneal epithelial cells were still alive under the action of 0.1% dihydroartemisinine. No effect on corneal epithelial cells under 0.025% dihydroartemisinine. The cells were dead under 0.006% chlorhexidine and 0.003% PHMB. Conclusion Dihydroartemisinine is hypotoxic for rabbit corneal epithelial cells in vitro and its effect on corneal cells in vivo remains to be further studied.