【作者】 王鹏；

【导师】 郭庆梅； 周凤琴；

【作者基本信息】 山东中医药大学 ， 中药学， 2011， 硕士

【摘要】 目的:运用溶剂法和多种色谱方法对白花丹参的化学成分进行提取分离和鉴定,探讨其化学成分对血管内皮细胞的保护作用,为白花丹参治疗血栓闭塞性脉管炎提供科学依据。方法:利用大孔树脂、硅胶和凝胶柱色谱等现代分离手段,对白花丹参脂溶性和水溶性化学成分进行分离和纯化,采用IR、MS、1H-NMR、13C-NMR等谱学方法对所得化合物结构进行鉴定。将体外培养的人脐静脉血管内皮细胞分为正常组、LPS凋亡损伤诱导组、白花丹参保护组,吖啶橙染色,共聚焦显微镜观测凋亡细胞核片段化状况;PI染核,流式细胞仪检测细胞周期分布。生化试剂盒检测细胞内脂质过氧化物(MDA)含量,超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)活性。结果:从白花丹参氯仿提取物中分离得到11个化合物,各种谱学技术鉴定了7个化合物,包括丹参酮ⅡA、隐丹参酮、丹参酮Ⅰ、二氢丹参酮Ⅰ、β-谷甾醇、丹参新醌乙、丹参素。白花丹参脂溶性部位及成分对LPS诱导的血管内皮细胞凋亡没有保护作用,水溶性部位及成分可能通过调节细胞氧化应激状态抑制脂多糖导致的血管内皮细胞凋亡。结论:本研究对白花丹参脂溶性和水溶性成分进行了提取分离,从中得到11个单体化合物。白花丹参脂溶性成分对血管内皮细胞有毒性作用,而其水溶性部位及成分对脂多糖处理的血管内皮细胞凋亡有一定的保护作用,其作用机制可能与调节细胞氧化应激状态有关。更多还原

【Abstract】 obliterans of Salvia miltiorrhiza f.alba(SAM), carry on the solvent extraction and various chromatographic methods to separate and identify its chemical composition, whose protective function to the blood vessel endothelial cell was discussed.Methods:Using the modern separation methods (pocket resin, silica gel and gelatin column chromatograph), separate and purify the fat-soluble and the water-soluble chemical composition of SAM, and identify their structure by IR, MS, 1H-NMR and 13C-NMR. carries on the appraisal to the obtained compound structure.Divided human umbilical vascular endothelial cell(HUVEC) in vitro into the normal control group, the LPS damage group, the SAM protection group; stained cells with acridinorange, and then observed cells under laser scanning confocal microcopy; stained cells with PI, and ananlyzed the cell cycle distribution with flow-cytometric The activities of glutathione peroxidase (GPH-PX) and Superoxide Dismutase (SOD) as well as maleic dialdehyde (MDA) level in cells were measured by biochemistry reagent box.Results:11 compounds were obtained from chloroform extraction of SMA, and 7 compounds were identified by various genealogy technology, including tanshinoneⅡA,cryptotanshinone,15,16-dihydrotanshinone,tanshinone,β-sitostenrol, danshixinkun B, danshensu. The fat-soluble ingredient and component of SMA had no protective function for the apoptosis of HUVEC induced by LPS, but the water-soluble ingredient and component was capable of protecting cells from apoptosis possibly through suppress the cell oxidation stress conditionion .Conclusion:This research extracted and separated the fat-soluble and the water-soluble chemical composition of SAM, and obtainsed 11 compounds.The fat-soluble chemical composition of SAM perfermed toxic effect to HUVEC, but its water-soluble chemical composition perfermed certain protective function, whose function mechanism is possibly concerned with regulating cell oxidation stress condition.更多还原