LC-MS测定两种来源续断中的川续断皂苷Ⅵ

目的 测定两种不同来源续断药材中不同部位的川续断皂苷Ⅵ.方法 采用LC-MS法和质谱法定性定量测定川续断皂苷Ⅵ的含量,选用Eclipse XDB C18柱(150 mm ×4.6 mm,5 μm),流动相为乙腈-水溶液(30:70),流速1.0 mL·min-1,检测波长210 nm;并用ESI-MS定性分析川续断皂苷Ⅵ.结果 川续断皂苷Ⅵ进样量0.204～2.04μg与峰面积的线性关系良好(r=0.9992);平均回收率为99.92%,RSD=1.31%.川续断药材中根、茎、叶中均含有川续断皂苷Ⅵ,含量分别为2.48%、0.60%、0.50%;而鹤峰续断只有根中含有川续断皂苷Ⅵ,含量为2.42%.结论 所建方法简便可行、专属性强,可控制续断药材及其制剂的质量.两种来源的续断植物在茎、叶中的含量差异可为川续断属植物提供化学分类学依据.     

茉莉酸甲酯对川续断根中川续断皂苷Ⅵ合成积累的影响

通过分析茉莉酸甲酯(MeJA)对川续断根中川续断皂苷Ⅵ积累及相关基因表达的影响,为深入研究MeJA调控川续断皂苷Ⅵ的合成奠定基础.本研究以川续断幼苗为实验材料,分别添加浓度为50、100、150、200、300 μmol·L-1的MeJA进行处理,在处理后1、3、5天不同时间点取样.分析根中川续断皂苷Ⅵ和超氧化物阴离子...     

川续断DaNAC基因家族成员的鉴定及表达分析

NAC基因在植物的逆境胁迫及次生代谢等过程具有重要的调控作用.为挖掘参与川续断皂苷Ⅵ合成的NAC基因家族成员,本研究利用川续断根、茎、叶、花、种子等不同组织构建的全长转录组数据库筛选包含完整开放阅读框的NAC家族成员,对其进行保守结构域及motif分析、进化树比对分析、组织表达差异分析和逆境胁迫诱导表达分析.本文共筛选...     

川续断皂苷Ⅵ药理作用研究进展

川续断皂苷VI又名木通皂苷D,是中国传统中草药川续断的主要活性成分。川续断皂苷VI药理作用广泛,其在神经保护、心肌保护、预防骨质疏松、抗细胞凋亡、镇痛等方面的药理活性引起了广大学者的关注,具有较高的研究和开发价值。本文就近年来有关川续断皂苷VI的药理作用研究进展作一综述。     

用大孔树脂纯化川续断中川续断皂苷Ⅵ的工艺研究

目的：研究用AB-8型大孔树脂富集纯化川续断中川续断皂苷Ⅵ的最佳工艺.方法：以川续断皂苷Ⅵ的动、静态吸附量和解吸附率为指标,考察了AB-8型大孔树脂对川续断中川续断皂苷Ⅵ的吸附和解吸附性能.结果：AB-8型大孔树脂纯化川续断皂苷Ⅵ的最佳工艺为：上样溶液的浓度为固含量0.08 g/ml,1g树脂上样量相当于1.2g生药材,先用30％乙醇除杂,再用70％乙醇解吸附,用量皆为3个柱体积,流速200 ml/h.得到富集浸膏的得率为8.93％,浸膏中川续断皂苷Ⅵ的含量为（65.32±1.73）％,转移率为95％.结论：该工艺能有效纯化、富集川续断药材中的川续断皂苷Ⅵ.     

不同产地续断的质量比较

目的 比较不同产地续断的质量.方法 采用HPLC法测定续断中川续断皂苷Ⅵ的含量,比色法测定续断总皂苷的含量.结果 中国14个不同产地续断的总皂苷含量为2.20%～19.91%,川续断皂苷Ⅵ含量为0.51%～10.14%.结论 各产地续断中川续断皂苷Ⅵ和总皂苷含量差异较大,以川鄂道地产区含量较高,可为药材的质量评价及发展道地药材的生产提供依据.     

续断壮骨胶囊中总皂苷和川续断皂苷VI含量测定

目的：建立续断壮骨胶囊中总皂苷和川续断皂苷遇的含量测定方法。方法：采用高氯酸显色,紫外-可见分光光度法测定总皂苷含量。采用高效液相色谱法测定川续断皂苷遇含量,Agilent C18柱（250 mm 215;4.6 mm,5μm）,乙腈-0.15%三氟乙酸水溶液（29∶71）为流动相,检测波长215 nm ,流量1.0 mL/min。结果：总皂苷平均含量为86.3%,平均加样回收率为99.88%, RSD =1.52%;川续断皂苷遇在64～320μg/mL 范围内呈良好线性关系（r =0.9995）,平均加样回收率为100.4%, RSD =0.77%。结论：所建方法准确,能快速测定续断壮骨胶囊总皂苷和川续断皂苷遇的含量,可用于该制剂的质量控制。     

续断配方颗粒中川续断皂苷Ⅵ含量比较

目的:测定不同厂家续断配方颗粒中川续断皂苷Ⅵ含量,为制定最佳提取工艺和统一的质量控制方法提供依据。方法:采用HPLC法测定,色谱柱为Diamonsil C18(2)(250 mm×4.6 mm,5μm),流动相乙腈-0.2%磷酸溶液梯度洗脱,检测波长212 nm,流速1 ml.min-1,柱温30℃。结果:川续断皂苷Ⅵ在0.603～3.016μg范围内呈良好的线性关系;供试品溶液在24 h内稳定,RSD为4.78%;加样回收率为99.49%(RSD=1.86%)。结论:所建立的方法适用于续断配方颗粒中川续断皂苷Ⅵ的含量测定。     

ＨＰＬＣ法测定续断接骨胶囊中川续断皂苷Ⅵ含量

目的　采用高效液相色谱法测定续断接骨胶囊中川续断皂苷Ⅵ的含量。方法　色谱柱：ＡｍｅｔｈｙｓｔＣ１８柱（４６ｍｍ×     

川续断皂苷Ⅵ不同给药方式对肺血栓小鼠的保护作用比较

目的 评估川续断皂苷Ⅵ不同的给药方式对治疗肺血栓的影响.方法 100只昆明小鼠分为尾静脉注射给药组和灌胃给药组,以川续断皂苷Ⅵ为治疗药物,随机将每组又分为空白组、模型组、尿激酶注射液/硫酸氢氯吡格雷组、川续断皂苷Ⅵ低剂量和高剂量组.以血栓诱导剂血栓形成法进行造模.以造模后15 min内的存活率、肺系数和6-keto-F...     

高效液相色谱法测定续断饮片中马钱苷的含量

目的建立中药饮片续断中马钱苷的含量测定方法。方法液相色谱法。色谱柱:InertsilRODS(4.6 mm×150 mm,5μm);流动相:乙腈-0.1%磷酸溶液(10∶90);流速1.0 mL·min-1;柱温:35℃;检测波长:240 nm。结果马钱苷在0.421 2～4.212μg范围内呈线性关系(r=1.000 0,n=6),平均加样回收率为98.9%,RSD为1.6%(n=6)。结论该方法操作简便,结果可靠,可用于中药饮片续断中马钱苷的含量测定。     

Treatment of Radix Dipsaci extract prevents long bone loss induced by modeled microgravity in hindlimb unloading rats

Abstract

Context: Radix Dipsaci is a kidney tonifying herbal medicine with a long history of safe use for treatment of bone fractures and joint diseases in China. Previous studies have shown that Radix Dipsaci extract (RDE) could prevent bone loss in ovariectomized rats.

Objective: This study investigates the effect of RDE against bone loss induced by simulated microgravity.

Materials and methods: A hindlimb unloading rat model was established to determine the effect of RDE on bone mineral density and bone microarchitecture. Twenty-four male Sprague–Dawley rats were divided into four groups (n?=?6 per group): control (CON), hindlimb unloading with vehicle (HLU), hindlimb unloading treated with alendronate (HLU-ALN, 2.0?mg/kg/d), and hindlimb unloading treated with RDE (HLU-RDE, 500?mg/kg/d). RDE or ALN was administrated orally for 4 weeks.

Results: Treatment with RDE had a positive effect on mechanical strength, BMD, BMC, bone turnover markers, and the changes in urinary calcium and phosphorus excretion. MicroCT analysis showed that RDE significantly prevented the reduction of the bone volume fraction, connectivity density, trabecular number, thickness, tissue mineral density, and tissue mineral content as well as improved the trabecular separation and structure model index.

Discussion and conclusion: RDE was demonstrated to prevent the loss of bone mass induced by HLU treatment, which suggests the potential application of RDE in the treatment of microgravity-induced bone loss.     

A subchronic toxicity study of Radix Dipsaci water extract by oral administration in F344 rats

Radix Dipsaci, the dried root of Dipsacus asperoides C.Y. Cheng & T.M.Ai, has therapeutic effects on various disorders, and in particular, bone and joint disease. Despite such ethnomedicinal benefits, there is very little information regarding its in vivo toxicity or adverse effects. This study was conducted to evaluate the potential toxicity of the Radix Dipsaci water Extract (RD-wE) by using F344 rats. The RD-wE was administered orally to rats at doses of 0, 125, 250, 500, 1000, and 2000 mg/kg body weight (bw)/day for 13 weeks. During the treatment period there were no mortalities attributed to RD-wE. Moreover, no toxic effects were observed with regard to body weight, clinical pathology (hematology, clinical biochemistry, and urinalysis), and anatomic pathology (gross findings, organ weight, and microscopic examination). The changes related to the treatment were excessive salivation at the mouth and soft feces, observed in male and female rats at 1000 or 2000 mg/kg bw/day, but these were not accompanied by any microscopic correlate or other pathophysiological changes. Based on these results, the oral no-observed-adverse-effect level of the RD-wE was considered to be 2000 mg/kg bw/day in both genders, although the target organs were not determined under the current experimental conditions.     

道地药材川续断指纹图谱模糊模式识别研究

目的建立道地药材川续断的指纹图谱测定方法。方法采用反相高效液相色谱法建立川续断指纹图谱，色谱柱为Shimadzu C18（150mm×4．6mm，5μm），流动相为甲醇一水梯度洗脱，流速为1．0mL／min，检测波长为210nm,柱温为25℃。结果建立了川续断指纹图谱标准。结论指纹图谱方法简便、快速、重现性好，可作为川续断的一项质量控制指标。     

续断高效毛细管电泳指纹图谱研究

目的:利用高效毛细管电泳(HPCE)方法建立中药续断的指纹图谱。方法:采用50μm×65 cm(有效长度57 cm)未涂层石英毛细管柱,以pH 9.20,50 mmol.L-1的硼砂-硼酸为缓冲溶液体系,运行电压25 kV,温度25℃,压力进样5 kPa×30 s,检测波长212 nm。采用"中药色谱指纹图谱相似度评价系统2004A版"软件和聚类分析方法,对样品图谱进行分析。结果:12批续断HPCE指纹图谱共确认了14个共有指纹峰,其中产自湖北的2批药材相似度介于0.8至0.9之间,产自四川的10批药材相似度大于0.9。聚类与相似度结果一致。结论:此法准确、可靠,可为续断药材的质量评价与控制提供科学依据。     

Quantitative determination and pattern recognition analyses of bioactive marker compounds from Dipsaci Radix by HPLC

In this study, quantitative and pattern recognition analyses were developed using HPLC/UV for the quality evaluation of Dipsaci Radix. For quantitative analysis, five major bioactive compounds were assessed. The separation conditions employed for HPLC/UV were optimized using ODS C₁₈ column (250 × 4.6 mm, 5 μm) with a gradient of acetonitrile and water as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 212 nm. These methods were fully validated with respect to linearity, accuracy, precision, recovery, and robustness. The HPLC/UV method was applied successfully to the quantification of five major compounds in the extract of Dipsaci Radix. The HPLC analytical method for pattern recognition analysis was validated by repeated analysis of 17 Dipsaci Radix and four Phlomidis Radix samples. The results indicate that the established HPLC/UV method is suitable for quantitative analysis.     

续断炮制历史沿革与现代炮制研究进展

目的介绍续断的炮制历史沿革与现代炮制的研究概况。方法查阅古籍和现代文献,总结历代炮制方法、炮制目的和现代炮制研究。结果与结论归纳总结出续断炮制的历史沿革和研究进展,为今后的研究方向提供了理论依据,为续断药材和饮片的质量与临床用药提供了安全和有效的保障。     

Advances in biotechnology for tissue engineering of bone

Tissue engineering is a rapidly developing field applying the disciplines of cell biology, developmental biology, molecular biology and biomimetic engineering to regenerate new tissues for replacement therapies in clinical contexts. To aid in the elicitation and reiteration of the processes of morphogenesis of tissues, the cascade of chemotaxis of progenitor cells, their differentiation and pattern formation is redeployed in postnatal tissues, effected by a variety of ever-increasing morphogens and biomaterials. The extensive recent progress in elucidating the molecular biology of BMPs and their receptors shall aid in promoting and extending the great operational future of this field. Although the BMP family of proteins and osteogenesis have been the subject of several recent reviews, we focus here on their activity in primates and on the novel localization of BMPs in the cerebellum and other areas of the nervous system, and the "mosaicism" of their localisation in the periodontal tissues followed by a discussion on the use of BMPs in periodontal regeneration. Lastly, we report on the unique osteoinductive activity of TGF-beta proteins in heterotopic sites of primates and their synergistic interaction with a recombinant human BMP, and finally we present unique data on novel biomaterials endowed with intrinsic osteoinductive activity, capable of initiating de novo bone formation in heterotopic sites even in the absence of exogenously applied BMPs, and the results of a clinical trial in humans using naturally-derived BMPs.     

续断炮制时间、饮片色度值及指纹图谱的相关性研究

目的 研究续断炮制时间、饮片色度值与UPLC指纹图谱的相关性。方法 建立续断饮片UPLC指纹图谱,监测酒续断、盐续断炮制过程中化学成分的变化,利用分光测色仪对续断不同炮制品炮制过程中饮片色度进行客观量化,运用SPSS 20.0、SIMCA 14.0软件分析炮制时间与饮片色度值和指纹图谱之间的相关程度。结果 酒续断、盐续断炮制过程中,饮片粉末颜色加深,粉末色度值(L^*、b^*、E^*)值均降低;通过相关性分析得出饮片炮制时间与饮片色度值和指纹图谱之间呈显著性相关。结论 建立的UPLC指纹图谱方法稳定、可靠,结合续断炮制品色度作客观判别分析,为更好地规范酒续断、盐续断炮制工艺及全面评价饮片质量奠定基础。     

Quantitative determination,principal component analysis and discriminant analysis of eight marker compounds in crude and sweated Dipsaci Radix by HPLC-DAD

Context: Dipsaci Radix is derived from the dry root of Dipsacus asper Wall.ex Henry (Dipsacaceae). It has attracted increasing attention as one of the most popular and precious herbal medicines in clinical use.

Objective: To develop a HPLC-DAD method for quantitative analysis and quality control of eight active components in crude and sweated Dipsaci Radix.

Materials and methods: The eight components in Dipsaci Radix were analyzed by HPLC-DAD on an Agilent Eclipse XDB-C18 column within a gradient elution of acetonitrile and 0.05% formic acid aqueous solution. ESI-MS spectra were acquired on a triple quadrupole mass spectrometer. Validation was performed in order to demonstrate linearity, precision, repeatability, stability, and accuracy of the method. The results were processed with principal component analysis (PCA) and discriminant analysis (DA).

Results: The eight components showed good linearity (R²?>?0.9991) in the ranges of 60.40–1208.00, 151.00–3020.00, 3.06–61.20, 30.76–615.20, 5.13–102.60, 10.17–203.40, 10.20–204.00, and 151.60–3032.00?mg/mL, respectively. The overall recoveries were in the range of 99.03–102.38%, with RSDs ranging from 1.89% to 4.05%. Through PCA, the degree of importance of the eight components in sequence was CA?>?AVI?>?IA?>?LA?>?LN?>?IC?>?IB?>?CaA. The crude and sweated Dipsaci Radix were distinguished obviously by DA.

Discussion and conclusion: The method, using HPLC-DAD analysis in combination with PCA and DA, could provide a more comprehensive and quantitative chemical pattern recognition and quality evaluation to crude and sweated Dipsaci Radix.