来氟米特对角质形成细胞增殖及凋亡的影响

Effects of Leflunomide on the Proliferation and Apoptosis of Human Keratinocytes

目的 研究来氟米特对角质形成细胞增殖及凋亡的影响。方法 以良性角质形成细胞株HaCaT为研究对象,用结晶紫染色及增殖细胞核抗原(PCNA)免疫组化法反映细胞增殖变化;HE染色观察药物处理前后细胞形态变化;流式细胞仪测定细胞周期及凋亡率变化。结果 5μmol/L来氟米特处理HaCaT细胞,24 h后表现出对上皮细胞增殖的抑制作用,PCNA阳性细胞开始减少。随着时间延长和药物剂量的加大,药物的抗增殖作用愈加明显,PCNA阳性细胞进一步减少,当药物浓度达到200μmol/L时,视野中未见PCNA阳性细胞。即药物对HaCaT细胞的抑制作用呈时间、剂量依赖关系。药物处理后,细胞变小或变圆,膜皱缩,核浓缩、深染,核分裂像减少。细胞S期比例增加,G2M期比例减少,G1期之前未见与药物剂量相关的凋亡峰。结论 来氟米特可抑制HaCaT细胞增殖,对细胞凋亡影响不大。

Objective To investigate the effects of leflunomide(A77 1726) on the proliferation and apoptosis of human keratinocytes. Methods Proliferating cell nuclear antigen (PCNA) of HaCaT cells was analyzed with crystal violet staining and immunohistochemistry. The moiphological change was assessed with hematoxylin and eosin staining. The changes of cell cycle and apoptosis rate were measured by flow cytome-try. Result Epidermal proliferation was inhibited by leflunomide, at concentration 5 μmol/L or more, which was begun after 24 hrs and manifested by PCNA positive cell decreasing. With the increasing of time or dosage, the anti- proliferation effect of leflunomide was significant. Meanwhile, the PCNA positive cell de-creased respectively. There was no PCNA positive cell while the drug concentration achieved 200 μmol/L. Therefore, leflunomide significantly inhibited the proliferation of HaCaT cells in a dose-dependent and time-dependent manner. The morphological change and cell cycle change was found but no change of apoptosis rate was measured. Conclusion Leflunmide can inhibit the proliferation of HaCaT cell, without the effect on its apoptosis.