雷公藤内酯醇的细胞内定位研究

In-cell localization of triptolide

目的检测雷公藤内酯醇的细胞内定位,探讨其在细胞内的作用位点。方法采用4-溴甲基-7-甲氧基香豆素标记雷公藤内酯醇,与人肝癌细胞孵育;并与细胞核染料PI或细胞质膜染料DiI行双色荧光染色,用荧光显微镜观察。结果与对照相比,香豆素标记在紫外激发下呈特异性的淡蓝色荧光;与DiI共定位显示,香豆素标记在细胞质(膜)存在;与PI共定位显示,香豆素标记在细胞核存在,且荧光强于细胞质(膜)。结论雷公藤内酯醇在细胞内主要位于细胞核,其次是细胞质(膜),提示雷公藤内酯醇可能主要在细胞核内起作用,其次是在细胞质(膜)上起作用。

Objective To clarify the cellular localization of triptolide and to explore its in-cell action sites. Methods 4-（/Sro- momethyl）-7-methoxycoumarin was employed to label triptolide,then labelled triptolide was incubated with human hepatoma carcinoma cells. Subsequently,incubated cells were subjected to stain with fluorescent dye DiI or PI,which were specific to cytoplasmic membrane system and nucleus,respectivel.y. Results Compared with the non-triptolide control,coumarin labelled triptolide shown a light blue fluorescence under UV excitation;Co-localization with DiI showed that triptolide exist in cytoplasm and（or） on cell membrane;Co-localization with PI showed that triptolide located in cell nucleus. Moreover, microscopic observation indicated that the fluorescence intensity in nucleus was denser than that in cytoplasm. Conclusion The presnt study demonstrate that triptolide main- lv act in nucleus,followed by acting in cytoplasm and（or） on cell membrane.