松果菊苷对大鼠成骨细胞胃桥素基因和蛋白质的影响

为了探讨松果菊苷对体外培养大鼠成骨细胞骨桥素(OPN)mRNA和蛋白质表达的影响,选取出生24 h内的SD大鼠颅盖骨,依次用胰酶和胶原酶消化分离培养成骨细胞,用不同浓度的松果菊苷作用于第3代细胞,选取雌二醇为阳性对照,分别在药物作用24、48和72 h后提取细胞总RNA的表达量,用荧光定量PCR(FQ-PCR)法检测细胞中OPN mRNA的表达量;用Western blotting检测细胞中OPN蛋白的表达量。结果显示,各浓度松果菊苷在各个时间段均增加OPN基因的表达,尤其是5×10^-5和5×10^-6 mg/mL松果菊苷在各时间段均具有显著或极显著地促进作用(P＜0.05或P＜0.01);各浓度组在作用48 h时蛋白质的表达量均高于对照组。结果表明,松果菊苷对成骨细胞OPN基因表达有显著的促进作用,同时对OPN蛋白的分泌有促进的趋势。     

Growth and accumulation of caffeic acid derivatives in Echinacea angustifolia DC. var. angustifolia grown in hydroponic culture

In separate experiments conducted in 2007 and 2008, growth and accumulation of selected caffeic acid derivatives (CADs; i.e., caftaric acid, chlorogenic acid, echinacoside, caffeic acid, cynarin, p-coumaric acid, ferulic acid and cichoric acid) were determined in Echinacea angustifolia DC. var. angustifolia seedlings grown in hydroponic culture (floating raft system) at a density of 122 plant m⁻² (at planting). Plants were harvested 11 (2007) or 16 (2008) weeks after transplanting (i.e., 15 or 20 weeks after sowing). In both years, plants grew vigorously and at harvest approximately half of the plants under observation had developed one to three inflorescences. In 2008, the root yield (2940 kg ha⁻¹) harvested in nearly eight months from two consecutive hydroponic cultures was within the yield reported in the literature for field cultivations lasting two to four years. None of the selected CADs was found in the leaves, while the inflorescences (stem and capitulum) contained only caftaric acid and echinacoside at concentrations higher than the detection limits (0.05 mg g⁻¹ dry weight). Echinacoside, cynarin and chlorogenic acid were found in root tissues at concentrations ranging from 0.36 to 5.25 mg g⁻¹ dry weight. In all plant samples, echinacoside, which is the marker compound for E. angustifolia material, did not reach the minimum quality standard (10 mg g⁻¹ dry weight) for the production of standardized extract. We concluded that short-cycle, high-density greenhouse hydroponic culture stimulates plant growth and root production in E. angustifolia, but it does not ensure sufficient CADs accumulation in dried roots.     

大孔树脂对管花肉苁蓉2种苷类静态吸附分离特性研究

松果菊苷和毛蕊花糖苷为管花肉苁蓉主要活性成分。以松果菊苷和毛蕊花糖苷为标准品,通过高效液相色谱仪确定样品质量浓度,从而比较8种供试树脂的吸附性能,以筛选出适宜分离此2种成分的大孔树脂。结果表明,X-5和HP-20吸附性能较好,进一步研究表明,X-5对松果菊苷吸附量最大,为50.03 mg/g,其次是HP-20,为46.05 mg/g;HP-20对毛蕊花糖苷的吸附量最大,为32.65 mg/g,其次是X-5,为27.56 mg/g。X-5对松果菊苷的解析能力最强,为77.35%,HP-20次之,为60.36%;X-5对毛蕊花糖苷的解析效果最好,为72.57%,其次是HP-20,为63.40%。2种树脂吸附量在前1 h变化最快,随时间的变化吸附能力逐渐减弱。X-5对试样原液(pH 3.43)中松果菊苷和毛蕊花糖苷的吸附性能优于HP-20,且当pH=6.02时对松果菊苷和毛蕊花糖苷的吸附量最大。确定X-5与HP-20分别为松果菊苷和毛蕊花糖苷的理想吸附树脂。综合比较吸附量和解析率等因素,确定X-5为分离纯化松果菊苷和毛蕊花糖苷的首选树脂。     

HPLC测定肉苁蓉微粉中松果菊苷和毛蕊花糖苷的含量

[目的]建立高效液相色谱法同时测定肉苁蓉微粉中松果菊苷和毛蕊花糖苷的含量.[方法]采用安捷伦1260高效液相色谱仪和Waters C18柱(4.6 mm×250 mm,5μm),以甲醇-0.1％甲酸水溶液为流动相梯度洗脱,流速1ml/min,检测波长330 nm,柱温30℃.[结果]松果菊苷和毛蕊花糖苷进样量分别在0.40～4.00、0.44～44.00μg范围内均与峰面积呈良好线性关系,平均回收率分别为97.4％和101.3％,RSD分别为1.70％、1.52％.[结论]该方法简单可行、重现性好、精密度高、结果可靠,可用于同时测定肉苁蓉微粉中松果菊苷和毛蕊花糖苷的含量.     

肉苁蓉中苯乙醇苷化合物的抗氧化活性研究

[目的]对内苁蓉药材中苯乙醇苷化合物的抗氧化活性进行系统分析研究。[方法]采用Photochemiluminescence（PCL）和2,2-diphenyl-1-picrylhydrazyl（DPPH·）2种不同的体系,测量肉苁蓉80％乙醇提取物以及麦角甾苷、异麦角甾苷、松果菊苷的抗氧化活性。[结果]松果菊苷总多酚的含量最高,其次为麦角甾苷、异麦角甾苷和肉苁蓉80％乙醇提取物,分别为753．95、659．94、356．14、14．73mg没食子酸／g。在PCL试验中,肉苁蓉80％乙醇提取物表现出比较强的抗氧化活性,其次为松果菊苷、麦角甾苷和异麦角甾苷;而在DPPH试验中,麦角甾苷表现出较强的抗氧化活性,其次为松果菊苷、异麦角甾苷和肉苁蓉80％乙醇提取物。[结论]该研究可为肉苁蓉药材中苯乙醇苷化合物的药理和生物活性研究提供参考。