青蒿琥酯对成纤维细胞和内皮细胞的选择性研究

目的: 探讨青蒿琥酯(Art) 是否对成纤维细胞增殖有特异的细胞选择作用。方法: 选用人胚肺成纤维细胞株(HLF) 、人皮肤瘢痕成纤维细胞(HSF) 和人脐静脉内皮细胞(HUVEC), 用免疫组化法鉴别细胞;MTT 法检测青蒿琥酯对上述3 种细胞存活与增殖的影响;光镜观察青蒿琥酯作用下细胞形态学的改变;琼脂糖凝胶电泳观察细胞DNA 的变化。结果: 青蒿琥酯在30 ~ 240 mg·L^-1浓度范围内作用细胞24 h 对两种来源的成纤维细胞的增殖都具有明显的抑制作用(P <0.05), 对血管内皮细胞增殖抑制作用不明显(P >0.05) 。药物作用24 h 后,HLF 、HSF 、HUVEC 细胞的IC₅₀ 分别为:64、60、600 mg·L^-1 。光镜下见两种成纤维细胞用药组细胞变圆, 突起变短变少, 胞浆颗粒增多;而内皮细胞在形态上无明显的变化。琼脂糖凝胶电泳结果显示在240 mg·L^-1浓度作用24 h, 两种成纤维细胞的DNA 出现梯带状的电泳图谱, 而内皮细胞的DNA 无梯状条带。结论: 青蒿琥酯对来源于肺和皮肤的成纤维细胞的增殖具有抑制作用, 而对血管内皮细胞生长增殖无明显的影响。该药还能诱导两种成纤维细胞凋亡, 而对脐静脉内皮细胞无明显诱导凋亡的作用。

Study of fibroblast apoptosis selectively induced by artesunate

AIM: To study the specificity of the antiproliferative effects of artesunate on fibroblast. METHODS: Human embryo lung fibroblasts cell line (HLF), human skin scar fibroblasts cell (HSF) and human umbilical vein endothelial cells (HUVEC) were selected in this trial. Cells were verified by immune his to-chemical method. A quantitative cytotoxicity was measured by MTT assay. Cell morphology change was identified by light microscopy. The apoptosis was evaluated by flow cytometry analysis and agarose gel electrophoresis. RESULTS: Artesunate inhibited the proliferation of HLF and HSF cells after cells were treated with artesunate for 24 hours in the dose range of 30-240mg·L^-1 (P <0.05). It had little inhibitory effects on HUVEC(P >0.05). Their IC₅₀ were 64, 60 and 600 mg·L^-1, respectively. Under light microscope, fibroblast treated with artemisinin assumed smaller, ellipse rather than fusiform in shape with increase of cytoplasmic particles, even with cell apoptosis. Ladder pattern of DNA were observed in artesumate-treated cells in the dose of 240 mg·L^-1.However, DNA of HUVEC cells was not affected in expropriate condition. CONCLUSION: The inhibitory effect of artesunate is specific on fibroblast coming from skin and lung. Artesunate can induce the apoptosis of the two fibroblasts, but it has no effect on HUVEC cells.