地黄多糖、车前子多糖、淫羊藿多糖及菟丝子多糖对成体大鼠肝脏干细胞生长影响的研究-中医骨伤科学专业论文.docxVIP

成都中医药大学硕 成都中医药大学硕士学位论文 PAGE PAGE 3 中文摘要 目的 ：通过观察同归肝肾经的四种中药的水溶性成分：地黄多糖、车前 子多糖、淫羊藿多糖及菟丝子多糖对成体大鼠肝脏干细胞生长、分化情 况的影响，从而更好地理解传统中药学归经理论，并验证“肝肾同源” 学说是否以药理学为基础。 方法： 将喂养含3’-甲基二甲胺基偶氮苯（3′- Me- DAB）饲料4周的SD大 鼠肝脏，通过灌注消化、梯度离心等方法制备成体大鼠肝脏干细胞。通 过分离纯化、传代增殖、倒置显微镜下观察细胞形态和生长情况。研究 不同浓度的地黄多糖、车前子多糖、淫羊藿多糖及菟丝子多糖对肝脏干 细胞的生长，增殖及其分化情况的影响。用四氮唑盐法(MTT比色法)监 测肝脏干细胞的生长活性，RT-PCR法检测经中药有效成分诱导后成体大 鼠肝脏干细胞的HNF4α mRNA的表达情况。 结果： MTT 比色法结果显示：淫羊藿多糖、菟丝子多糖在特定的浓度 下能提高肝脏干细胞活性，促进细胞增殖；地黄多糖、车前子多糖在特 定浓度区间内不能促进细胞生长及增殖。RT-PCR 结果显示：经特定浓 度下的淫羊藿多糖、菟丝子多糖诱导后，细胞的 HNF4α mRNA 表达明 显增强。 结论：（1）特定浓度下淫羊藿多糖、菟丝子多糖可促进成体大鼠肝脏干 细胞的增殖。在一定浓度区间内地黄多糖、车前子多糖对肝脏干细胞的 增殖无影响。提示性寒凉的“中药多糖”对细胞的生长和代谢无明显的 促进作用。（2）经淫羊藿多糖、菟丝子多糖环境诱导后，细胞的 HNF4α mRNA 表达明显增加，提示在性温热的“中药多糖”的诱导下，肝脏干 细胞生长活性增强的同时，其向肝脏细胞的分化趋势也十分明显。 关键词 ：地黄多糖 车前子多糖 淫羊藿多糖 菟丝子多糖 干细胞 肝肾 同源 Abstract Objective: Rehmannia polysaccharide, Plantago polysaccharide, Epimed -ium polysaccharide, dodder seed polysaccharide are all water-soluble components attributed to the liver and kidney meridian, by observing their effects on the growth and differentiation of Adult rat liver stem cell. we can understand the meridian theory of traditional Chinese medicine and verify the theory of the liver and kidney having a common source whether take the Pharmacology as the rationale. Methods: SD rats fed with 3-methyl-dimethylamino azobenzene (3-Me- DAB) for 4 weeks, we prepare SD rat liver stem cell from the liver by perfusion digestion, gradient centrifugation method. Through removing non-adherent cell via renewing culture fluid, we can separate and purify the stem cells and then, made them passage and proliferate. Cell morph- ology was observed under inverted microscope so as to acquire the rule of cell metabolism. Then we observed the effects of different concentrations of Rehmannia polysaccharide, Plantago polysaccharide, dodder seed polysaccharide on liver stem cell growth, proliferation and apoptosis, applying tetrazolium salt (MTT Cell Proliferation As-say, MTT assay) to monitor activity of liver stem cells; an