基于qRT-RNA技术筛选藏药湿生扁蕾抗溃疡性结肠炎肠纤维化活性成分的研究

摘要

Objective To study the bioactive constituents of Tibetan medicine Gentianopsis paludosa against UC fibro-sis.Methods Active fragment were obtained by silica gel and TLC from ethyl acetate active part which was extracted from Tibetan Gentianopsis paludosa.By method of qRT-PCR,collagen Ⅰ,collagen Ⅲ,α-SMA,and E-cad were detected to screen the active components after model rats being treated with active isolates.Results After purified and identified from ethyl acetate part using spectroscopy,four monomer compounds(1-hydroxy-3,7,8-Trimethoxyxanthones;1,8-Dihydroxy-3,7-Dimethoxyxanthones;1-hydroxy-3,7-Dimethoxyxanthones and 1,7-Dihydroxy-3,8-Dimethoxyxanthones)had sig-nificant bioactive on UC fibrosis.Conclusion The material basis of Gentianopsis paludosa against fibrosis concentrated in parts of ethyl acetate,and the active ingredient mainly owed to four xanthones.%目的：研究藏药湿生扁蕾抗溃疡性结肠炎肠纤维化的药效物质基础。方法采用2，4，6-三硝基苯磺酸（TNBS）诱导的大鼠溃疡性结肠炎肠纤维化模型，灌胃给予藏药湿生扁蕾用乙酸乙酯萃取的活性提取物，以硅胶色谱分离、薄层定性得到的活性部位，采用荧光定量 RT-PCR 法检测大鼠结肠组织胶原ⅠmRNA、胶原Ⅲ mRNA、α-SMA mRNA、E-cad mRNA 的表达，筛选其活性成分。结果湿生扁蕾乙酸乙酯萃取物分离得到的活性部位经纯化得到4个单体化合物，用光谱分析等鉴定为1-hydroxy-3，7，8-Trimethoxyxanthones、1，8-Dihydroxy-3，7-Dimethoxyx-anthones、1-hydroxy-3，7-Dimethoxyxanthones 和1，7-Dihydroxy-3，8-Dimethoxyxanthones，4种单体化合物对溃疡性结肠炎肠纤维化大鼠模型均有明显的活性。结论湿生扁蕾抗溃疡性结肠炎肠纤维化的物质基础集中在乙酸乙酯部位，活性成分主要体现为4种酮类化合物。