四物汤补血调血作用机理的研究

通过测定血清红细胞生成素（ＥＰＯ）和脾条件培养液（ＳＣＭ）中红细胞生成素样生成因子的水平,研究四物汤对红系细胞补血调血的作用机理。实验结果显示四物汤能显著提升正常小鼠、正常和血虚大鼠血清ＥＰＯ水平,及能显著提长正常和血虚大鼠ＳＣＭ中ＥＰＯ样生成因子水平,提示四物汤对红系细胞的补血调血作用机理及其刺激ＥＰＯ分泌和促进ＥＰＯ样生长因子产生有关。     

四物汤中糖类物质对辐射小鼠造血功能的影响

目的初步探讨四物汤中糖类物质与四物汤补血作用的关系。方法四物汤部位分离采用醇沉法、萃取法和大孔吸附树脂法。用辐射小鼠血虚证模型考察活性。结果将前期发现的具有提高辐射血虚模型小鼠外周血白细胞数和促进骨髓造血祖细胞增殖作用的四物汤部位C 2(正丁醇部位),用吸附树脂法分离为C 2-1(糖部分)和C 2-2(非糖部分)两个部位。活性研究表明部位C 2-1具有提高模型小鼠外周血白细胞数和促进骨髓造血祖细胞增殖的作用,部位C 2-2仅显示促进骨髓造血祖细胞增殖的作用。按照部位C 2中糖类的组成和质量分数,将D-果糖、D-葡萄糖和蔗糖混合,活性研究表明混合糖具有促进模型小鼠骨髓造血祖细胞增殖的作用,但对外周血白细胞的作用不显著。结论单、双糖类物质对四物汤促进辐射小鼠造血功能的作用有一定贡献。     

四物汤的方证结合研究

方证结合研究是中医理法与方药之间的桥梁,以经典补血方剂四物汤与血虚证的方证关系研究为例探讨方证结合研究的新模式。运用射线照射、环磷酰胺腹腔注射和综合放血法复制动物模型,研究发现血虚证的发生发展与外周血各血液成份改变、骨髓造血干/祖细胞的动态变化、骨髓细胞凋亡、CD34+细胞的改变、细胞周期、造血相关基因和蛋白的改变等密切相关。进而从全方、有效部分、有效成分3个化学层次和整体、器官、细胞及分子4个药理水平,从方证到理法的逆向辨证探求方剂与证候间的内在联系,基本阐明了补血名方四物汤的药效物质基础,并重构了基于有效成分的新组方,首次发现了果糖和芍药苷的补血活性,研究体现了中医药方证结合研究的特色和优势,为方剂和证候学的结合研究提供了参考模式。     

四物汤补血有效部位及对红系特异性转录因子GATA-1 mRNA表达的影响

目的探讨四物汤补血有效部位及对红系特异性转录因子GATA-1 mRNA表达的影响。方法采用极性由小到大的三种溶剂石油醚、醋酸乙酯及水分别提取四物汤组分，用半定量RT—PCR分析红系转录因子GATA-1 mRNA表达水平。结果与血虚小鼠模型组及石油醚提取物给药组比较，“四物汤”水提物能明显增强血虚小鼠模型红系造血，亦能显著上调红系转录因子GATA-1 mRNA的表达（P〈0．01）。结论“四物汤”水提物系补血有效部位，GATA一1mR_NA过表达可能在介导“四物汤”促红系造血中起重要作用。     

Hemopoietic effect of extracts from constituent herbal medicines of Samul-tang on phenylhydrazine-induced hemolytic anemia in rats

Samul-tang (Si-Wu-Tang, SMT), a kind of herbal medicines, has been used for the hemato-deficient disease for hundreds of years. In this work, investigate the anti-anemia activity of the H₂O extracts from constituent herbal medicines of Samul-tang in an anemia model induced by intravenous infection of phenylhydrazine-HCL (PHZ) at 10 mg/kg for 4 days. After PHZ injection, female Sparague-Dawley rats were administrated extracts from constituent herbal medicines of SMT (300 mg/kg/day, p.o.) daily for 1 week. Results showed that sever hemolysis was induced by PHZ. For Paeonia lactiflora (PL2) H₂O extract treated groups, the concentration of hemoglobin, hematocrit and red blood cells number increased much more significantly than PHZ-treated group. Moreover, Angelica gigas (AG), Angelica. acutiloba (AA), Paeonia lactiflora (PL2) and Rehmannia glutinosa (RG) extract administration significantly improved serum erythropoietin concentration. The activity of aminolevulinic acid dehydrates (ALDL) in liver homegenate was increased in Angelica gigas(AA), Paeonia lactiflora (PL2) and Rehmannia glutinosa (RG) treated group.     

基于网络药理学整合体内实验探究四物汤单味药组分治疗胆汁淤积性肝损伤的作用与机制

目的 基于网络药理学整合体内实验对比四物汤中4种单味药（川芎Chuanxiong Rhizoma、白芍Paeoniae Radix Alba、当归Angelicae Sinensis Radix、熟地黄Rehmanniae Radix Praeparata）治疗胆汁淤积性肝损伤（cholestatic liver injury,CLI）的药效差异,并探讨各单味药的作用机制。方法 利用TCMSP数据库获取川芎、白芍、当归、熟地黄的主要活性成分及作用靶点,通过Gene Cards数据库获取CLI靶点,使用Cyctoscape 3.10.1软件分析并构建“单味药-活性成分-作用靶点”网络,借助在线网站获取各单味药与CLI之间的交集靶点,并绘制韦恩图。将靶点基因导入DAVID数据库进行基因富集分析。构建胆总管结扎（bile duct ligation,BDL）动物模型,给予各单味药水提物后,检测血清肝功能相关生化指标,并对肝组织进行病理染色对比药效作用。同时结合qRT-PCR检测CLI相关靶点的基因表达。结果 网络药理学结果显示,四物汤中川芎、白芍、当归、熟地黄与CLI之间均存在3个交集靶点[...     

Simultaneous quantification of active components in the herbs and products of Si-Wu-Tang by high performance liquid chromatography–mass spectrometry

Si-Wu-Tang (SWT), comprising Paeoniae, Angelicae, Chuanxiong and Rehmanniae, is one of the most popular Traditional Chinese Medicine (TCM) formulae for woman's health. Data mining from the available Chinese and English literatures indicated that the major bioactive components of SWT consist of paeoniflorin, paeonol, gallic acid, ferulic acid, Z-ligustilide, ligustrazine, butylphthalide, senkyunolide A and catalpol. Since content determination of the marker compounds is generally considered as an initial step for quality control of TCM product, a high performance liquid chromatography–mass spectrometric method employing both positive and negative electrospray ionization was developed for the simultaneous determination of the nine identified compounds in the raw herbs and products of SWT. The LOQ of the developed assay method for the tested components was 10 ng/ml for ligustrazine, 200 ng/ml for catalpol, and 100 ng/ml for the other seven compounds. The intra-day and inter-day variations of the current assay were within 17.5%. Paeoniflorin, ferulic acid, gallic acid, Z-ligustilide and senkyunolide A were found in all SWT products investigated. Variations in the contents of the studied compounds were observed among batches of raw herbs and SWT products. The currently developed method provides a sensitive and rapid quantification approach that can be useful in the quality control of raw herbs and products of SWT.     

四物汤对辐射致血虚证小鼠血清蛋白质表达的影响

目的 :用蛋白质组技术考察四物汤对血虚证小鼠血清蛋白的影响 ,在分子水平上探讨四物汤补血的作用机理。方法 :⁶⁰Coγ射线全身照射小鼠造成血虚证模型 ;二向电泳、图象分析、质谱鉴定等蛋白质组学技术测定四物汤对血虚证小鼠血清中有影响的蛋白。结果 :四物汤可使血虚证小鼠血清中 12个下调和 4个上调的蛋白质有所恢复 ,进一步质谱鉴定其中 4个蛋白质可能是DNA依赖蛋白激酶 (DNA -dependent protein kinase catalytic sub unit,P973 13 ) ,肌细胞增强蛋白 (dystrophin ,P11531) ,马达蛋白 (KIF13A ,Q9EQW7) ,肌动蛋白结合蛋白 (dys-tonin ,Q9QX20 )。它们的功能是参与DNA双链损伤的修复 ,与AP-1结合将 6-磷酸甘露糖受体从高尔基体转运到胞浆膜上 ,将细胞骨架铆定在胞浆膜上等。结论 :四物汤可能通过对血虚证小鼠血清中蛋白的影响而促进骨髓造血、减轻辐射引起的损伤。     

四物汤中糖类物质对辐射小鼠造血功能的影响

目的初步探讨四物汤中糖类物质与四物汤补血作用的关系。方法四物汤部位分离采用醇沉法、萃取法和大孔吸附树脂法。用辐射小鼠血虚证模型考察活性。结果将前期发现的具有提高辐射血虚模型小鼠外周血白细胞数和促进骨髓造血祖细胞增殖作用的四物汤部位C 2(正丁醇部位),用吸附树脂法分离为C 2-1(糖部分)和C 2-2(非糖部分)两个部位。活性研究表明部位C 2-1具有提高模型小鼠外周血白细胞数和促进骨髓造血祖细胞增殖的作用,部位C 2-2仅显示促进骨髓造血祖细胞增殖的作用。按照部位C 2中糖类的组成和质量分数,将D-果糖、D-葡萄糖和蔗糖混合,活性研究表明混合糖具有促进模型小鼠骨髓造血祖细胞增殖的作用,但对外周血白细胞的作用不显著。结论单、双糖类物质对四物汤促进辐射小鼠造血功能的作用有一定贡献。