正交试验法优选更年安舒颗粒提取工艺

摘 要 目的： 优选更年安舒颗粒的提取工艺。方法: 采用正交试验设计法,以松果菊苷及特女贞苷含量为指标综合评价,考察加水量、提取时间、提取次数对提取工艺的影响。结果: 最佳提取工艺为加水煎煮2次,每次2 h,每次加8倍量的水。结论: 优选的工艺合理可行、重复性良好、提取效率高,适合于工业化生产。     

不同初加工温度对肉苁蓉有效成分含量的影响

目的:考察不同初加工温度对内苁蓉中主要有效成分含量的影响。方法:以333nm为测定波长,以松果菊苷为对照品,采用大孔吸附树脂-UV法测定不同初加工温度下肉苁蓉中苯乙醇苷的含量;采用2005年版《中国药典》"肉苁蓉"项下HPLC法,测定不同初加工温度下肉苁蓉苯乙醇苷中主要成分松果菊苷和毛蕊花糖苷的含量;采用硫酸-苯酚法,测定不同初加工温度下肉苁蓉中多糖的含量。结果:苯乙醇苷的含量以自然晾干和干燥箱内40℃干燥的结果为高;随着初加工温度的升高,苯乙醇苷的含量大部分有下降的趋势。松果菊苷和毛蕊花糖苷的总量以自然晾干为最高,但不呈规律性的变化。多糖的含量以干燥箱内40℃和60℃干燥的结果为高,其中以60℃干燥的结果为最高;其后,随着初加工温度的升高,多糖的含量下降。春季药材上部(有少许中空)苯乙醇苷的含量低于药材下部的含量;且上部松果菊苷和毛蕊花糖苷的总量大大低于药材下部的含量,最大相差约27倍;多糖的含量也以药材下部为高。秋季药材中的多糖含量比春季药材(上部有少许中空)的含量高。结论:本研究结果为肉苁蓉的初加工方法提供了科学依据。     

松果菊苷通过激活ERK/BMP-2信号通路促进大鼠的成骨细胞增殖观察

目的：考察松果菊苷对大鼠成骨细胞增殖的影响及作用机制。方法将不同浓度的松果菊苷（0.01,0.1,1.0,10.0,100 nmol/L）及MAPK/ERK信号通路抑制剂PD98059（20μmol/L）与成骨细胞共培养12,24,36,48,60 h后,采用MTT法检测细胞增殖情况。酶联免疫吸附法（ELISA）和qRT-PCR法检测细胞上清中和成骨细胞中骨钙素（BGP）和骨形态发生蛋白（BMP-2）的表达水平。Western blotting检测成骨细胞中Smad4、ERK1/2和p-ERK1/2的蛋白表达水平。结果松果菊苷能够促进大鼠成骨细胞的增殖,而且能诱导ERK 的磷酸化从而激活 MAPK/ERK 信号通路,并提高成骨细胞中 BMP-2和 Smad4的表达而激活BMP/Smad信号通路。加入PD98059后,松果菊苷诱导的BMP-2和Smad4表达及成骨细胞增殖均受到抑制。结论松果菊苷可通过激活ERK/BMP-2信号通路促进大鼠的成骨细胞增殖。     

Application of 1H NMR-based metabolomics for discrimination of different parts and development of a new processing workflow for Cistanche deserticola

Cistanche deserticola (CD) is one of the two authoritative source plants of Cistanches Herba, a well-known medicinal plant. Herein, ¹H NMR spectroscopy was employed to characterize the chemical profile and to distinguish the different parts, as well as to propose a new processing workflow for CD. Signal assignment was achieved by multiple one and two dimensional NMR spectroscopic techniques in combination with available databases and authentic compounds. The upper parts of the plant were distinguished from the lower parts by combining ¹H NMR spectroscopic dataset with multivariate statistical analysis. A new processing method that hyphenated steaming with freeze-drying, was demonstrated to be superior to either steaming coupled with oven-drying or direct freeze-drying via holistic ¹H NMR-based metabolomic characterization. Phenylethanoid glycosides, mainly echinacoside and acteoside, were screened out and confirmed as the chemical markers responsible for exhibiting the superiority of the new processing workflow, whereas serial primary metabolites, especially carbohydrates and tricarboxylic acid cycle metabolites, were found as the primary molecules governing the discrimination between the upper and lower parts of the plant. Collectively, ¹H NMR spectroscopy was demonstrated as a versatile analytical tool to characterize the chemical profile and to guide the in-depth exploitation of CD by providing comprehensive qualitative and quantitative information.     

高效液相色谱法测定鱼鳔补肾丸中松果菊苷的含量

目的:建立高效液相色谱法测定鱼鳔补肾丸中松果菊苷的含量.方法:采用shim-pack CLC-ODS(6 mm × 150 mm,5μm)色谱柱,流动相:甲醇-0.1%甲酸溶液(27:73),流速1.0 mL/min,柱温35℃,进样量10μL,检测波长334nm.结果:松果菊苷在进样量0.01017～0.05085 mg/mL范围内线性关系良好,回归方程为Y=976220X-2600.8(r=0.9998,n=5),平均加样回收率为101.09%,RSD为1.75%.结论:本方法快速、简便、结果准确,可用于控制该制剂的质量.     

HPLC法测定管花肉苁蓉中松果菊苷和麦角甾苷的含量

目的：建立同时测定管花肉苁蓉药材及提取物中松果菊苷和麦角甾苷含量的HPLC法。方法：采用Hypersil ODS C18（200mm×4．6mm,5μm）色谱柱,甲醇-乙腈-1％醋酸溶液（15：10：75）为流动相,流速为0．8ml·min^-1,检测波长为334nm,柱温为30℃。结果：松果菊苷和麦角甾苷线性范围分别在3．65—234．00μg·ml^-1（r=0．9999）和2．74～175．20μg·ml^-1（r=0．9999）浓度范围内呈良好的线性关系,平均回收率分别为95．7％,95．8％,RSD分别为1．8％,2．7％（n=9）。结论：方法简便,精密度、重复性良好,结果准确可靠,可用于工业生产肉苁蓉药材、饮片及其制剂的含量测定。     

松果菊苷通过下调p53的表达抑制人成纤维细胞的衰老(英文)

松果菊苷是一种从肉苁蓉中分离得到的苯乙醇苷类化合物,前期研究结果表明其有延缓人成纤维细胞衰老的作用。为了阐明松果菊苷抗衰老的机制,我们对相关基因p53,p16,p21及Rb的mRNA及蛋白水平的表达进行了检测,结果表明用松果菊苷处理后,衰老的人成纤维细胞(MRC-5)p53的表达被下调,且呈剂量依赖方式。进一步的研究显示可能和SIRT1白的上调有关。分子对接模拟结果显示松果菊苷的作用可能优于另一公认的抗衰老剂白藜芦醇。松果菊苷可能是一种潜在的可以调控细胞衰老的化合物。     

肉苁蓉类生药中苯乙醇甙类成分的 RP-HPLC 分析

采用反相高效液相色谱法,对4种及1变种肉苁蓉类生药和25份商品药材所含的苯乙醇甙类成分进行了定性和定量分析,结果表明,荒漠肉苁蓉 Cistanche deserticola Ma,盐生肉苁蓉 C.salsa(C.A.Mey) G.Beck,白花盐苁蓉C.salsa var.albiflora P.F.Tu et Z.C.Lou和管花肉苁蓉C.tubulosa (Schenk) Wight 所含成分相似,沙苁蓉 C.sinensis G.Beck区别较大;松果菊甙(echinacoside)和类叶升麻甙(acteoside)的含量以盐生肉苁蓉最高,分别为2.13%和1.51%。采用Altima C₁₈,5μm,250×4.6mm色谱柱;乙腈─1.5%乙酸水溶液,其中乙腈浓度从8%→20%,0～60min线性梯度洗脱,作为定性分析流动相,乙腈浓度从11.5%→20%,0～35min线性梯度洗脱,作为定量分析流动相;流速1.2ml·min^-1,UV335 nm检测。     

松果菊苷对局灶性脑缺血大鼠脂质过氧化损伤的影响

目的探究松果菊苷（ECH）对局灶性脑缺血大鼠脑组织匀浆中谷胱甘肽（GSH）、超氧化物歧化酶（SOD）和丙二醛（MDA）的影响。方法将SD大鼠随机分为假手术组、模型组、ECH高剂量组、ECH低剂量组、阳性药物组,并给予相应药物,7d后采用改良线栓法制作左侧大脑局灶性脑缺血模型（MCaO模型）,术后24h断头取脑,制备脑匀浆,用生化方法检测GSH、MDA及SOD的含量。结果与模型组比较,ECH高剂量组、ECH低剂量组和阳性药物组脑组织中SOD和GSH活性明显升高,MDA含量明显降低,差异有统计学意义（P〈0．05或P〈0．01）。结论ECH具有一定的抗氧化作用。     

Echinacoside prevents the striatal extracellular levels of monoamine neurotransmitters from diminution in 6-hydroxydopamine lesion rats

We investigated the effects of echinacoside, a phenylethanoid glycoside isolated and purified from the stems of Cistanche salsa, a Chinese herbal medicine, on the striatal extracellular levels of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in 6-hydroxydopamine (6-OHDA) lesion rats. Seven days after 6-OHDA was injected into the right striatum of rats, the striatal extracellular levels of DA, DOPAC and HVA fell significantly (P<0.01 vs. vehicle), as demonstrated by the method of cerebral microdialysis and high performance liquid chromatography with electrochemical detection. However, simultaneous treatment with echinacoside (7.0, 3.5mg/kg) attenuated the diminution of them (P<0.01 vs. model). The results implied that echinacoside could protect the striatal dopaminergic neurons from injury induced by 6-OHDA and may be useful in the prevention and treatment of Parkinson's disease (PD).