制何首乌多糖的超声提取工艺

目的 :优选超声提取制何首乌中多糖的最佳工艺。 方法 :采用正交设计的方法,考察提取时间、提取温度、料液比、提取次数对制何首乌多糖得率的影响,优选出最佳的超声提取工艺,并与热水浸提法进行比较。 结果 :超声提取的最佳条件为用25倍的水,70 ℃超声提取,提取时间50 min,提取2次,多糖的得率20.04%,与热水浸提法相比提取时间明显减少。 结论 :该方法快速、高效、可行。     

《中华人民共和国药典》2020年版收载何首乌和制何首乌质量标准【鉴别】项的商榷

目的 讨论完善《中华人民共和国药典》（以下简称《中国药典》）2020年版何首乌及制何首乌的薄层色谱鉴别项。方法 考察超声及加热回流2种不同的样品制备方式,以及用二氯甲烷-甲醇-甲酸、乙酸乙酯-石油醚-甲酸展开系统代替原标准中三氯甲烷-甲醇展开系统的可行性,同时比较了不同显色条件。结果 2种制备方式制得的样品在相同的展开条件下展开结果并没有明显差异,超声提取操作更加简便;以二氯甲烷-甲醇-甲酸（5∶1∶0.3）和石油醚-乙酸乙酯-甲酸（5.0∶1.0∶0.3）代替三氯甲烷-甲醇展开系统后,喷10%硫酸乙醇显色剂并在紫外灯365 nm下检视,薄层色谱板上斑点明显增多且分离效果好,较《中国药典》2020年版何首乌和制何首乌薄层色谱鉴别条件更理想。结论 改进后的方法毒性低、简便易行、可行性良好,为进一步完善《中国药典》2020年版何首乌和制何首乌质量标准提供参考。     

制何首乌多糖的超声提取工艺

目的:优选超声提取制何首乌中多糖的最佳工艺。方法:采用正交设计的方法,考察提取时间、提取温度、料液比、提取次数对制何首乌多糖得率的影响,优选出最佳的超声提取工艺,并与热水浸提法进行比较。结果:超声提取的最佳条件为用25倍的水,70℃超声提取,提取时间50 min,提取2次,多糖的得率20.04%,与热水浸提法相比提取时间明显减少。结论:该方法快速、高效、可行。     

液相微萃取 - 高效液相色谱法测定制首乌中蒽醌类化合物

 目的 中空纤维液相微萃取结合高效液相色谱法同时对制首乌中蒽醌类化合物作定量分析。 方法 利用自制的液相微萃取装置,以聚偏氟乙烯纤维（ MOF503 ）为溶剂载体,正辛醇为萃取溶剂,供相中 HCl 浓度为 2 mmol·L^-1 ,供相中甲醇比例为 50% ,搅拌速度为 1 800 r·min^-1 ,萃取时间为 60 min 。萃取结束,萃取液经 80 ℃ 水浴蒸干后用 60 μL 甲醇溶解,在 434 nm 处进行 HPLC 分析 。 结果 在优化的液相微萃取条件下, 芦荟大黄素 、大黄酚、大黄酸、大黄素甲醚和大黄素检测限分别为： 0.30 , 0.25 , 0.25 , 0.35 和 0.30 μg·L^-1 ,精密度在 8.1% ～ 14.1% 之间,在制首乌中的回收率在 75.4% ～ 111.5% 之间。 5 种化合物液相微萃取富集倍数分别为 21 , 21 , 36 , 44 , 47 倍。测定了制首乌中大黄素、大黄素甲醚和痕量大黄酸、大黄酚的含量 。 结论 液相微萃取是一种简单、快速、选择性高、富集倍数高、有机溶剂消耗少的样品前处理技术,可用于同时分析制首乌药材中蒽醌类化合物 。     

结合传统性状客观化分析何首乌不同炮制方式与炮制程度的色彩色差

目的:结合传统性状观察,对不同炮制方式下不同炮制程度的制何首乌进行色彩色差分析,为何首乌饮片颜色的客观数据化提供研究基础。方法:观察何首乌炮制品性状,采用CM-5型测色仪测定样品的L,a,b,计算色差值ΔE与总色值E,运用SAS 9.2统计软件进行聚类分析,分析炮制程度与色差检测结果的相关性。结果:2个产地的何首乌生品及其对照饮片粉末均为黄白色,E为70.31±3;制何首乌对照饮片粉末为棕褐色,E为52.24,与生何首乌对照饮片比较,ΔE为18.63;何首乌炮制后颜色加深,E降低,4种炮制方式显示相同的变化规律;且随炮制时间延长,颜色逐渐由棕黄色、浅棕色、深棕色、棕褐色直至变为棕黑色,其E也由61.11逐渐降至30.45,说明总色值能够表明何首乌的炮制程度。通过聚类分析可区分生品与炮制品,并将不同炮制程度的炮制品聚为一类。结论:色差仪的检测数据可以准确区分生何首乌与制何首乌,且能够客观准确地表达不同炮制程度的制何首乌粉末颜色变化。色彩色差检测可用于何首乌饮片的质量评价。     

何首乌炮制后化学成分及药理作用的研究进展

何首乌作为我国传统中医常用的滋补药,在临床上应用很广泛,尤其是补肝方面,但是最近几年临床上一直有关于何首乌生品及炮制品引起肝损伤的不良反应产生,引起了研究人员的关注。本文就其总结了近年来关于何首乌生品及炮制品的相关报道,从何首乌的炮制历史沿革、炮制工艺、炮制对药物化学成分变化和药理作用的影响等方面进行了总结和归纳。何首乌生品及炮制品的化学成分、药理作用方面均存在一定的差异,考虑到临床用药的安全性、有效性及中成药的质量稳定性,在使用过程中应区分使用并注意合理用药,为正确应用何首乌提供参考依据。     

制首乌4种炮制过程中物质基础共性变化规律分析

为了规范制首乌的炮制工艺,稳定制首乌饮片的质量,该研究基于物质基础内涵对不同炮制方法及不同炮制时间制首乌进行了共性变化规律分析,考察了黑豆汁炖、清蒸、黑豆汁蒸和黑豆汁黄酒蒸4种炮制方法制首乌,在炮制24,32,40,48 h时,二苯乙烯苷类、蒽醌类、多糖、寡糖类等9种主要化学成分的含量变化。采用HPLC-DAD测定二苯乙烯苷类及蒽醌类成分的含量;紫外-可见分光光度法测定多糖类成分含量;HPLC-ELSD测定D-果糖、D-无水葡萄糖与蔗糖含量,绘制含量测定结果比较图,结合聚类分析和熵权TOPSIS模型,寻找不同炮制方法制首乌炮制过程中物质基础共性变化规律时间区间。结果显示,炮制32 h左右,4种不同炮制方法制首乌各主要成分含量具有一定共性特点且具有较高的质量。因此,将不同炮制方法制首乌炮制过程中物质基础变化规律相似时区设置为32 h左右。该研究表明4种炮制方法具有"统一"的可能性,研究结果为不同炮制方法制首乌作为同名饮片使用的科学性提供了参考。     

基于过程控制的何首乌产地加工与炮制一体化方法分析

目的:比较产地加工与炮制一体化方法和传统炮制方法对何首乌中二苯乙烯苷、游离蒽醌类成分和总多糖含量的影响,为建立何首乌产地加工与炮制一体化方法提供依据。方法:采用一体化方法和传统炮制方法对何首乌饮片清蒸和黑豆汁拌蒸,在不同时间点取样,分别测定二苯乙烯苷、游离蒽醌类(大黄素、大黄素甲醚)和总多糖的含量。采用HPLC测定二苯乙烯苷和游离蒽醌类成分的含量,二苯乙烯苷含量测定的流动相为乙腈-水(25∶75),检测波长320 nm;游离蒽醌类成分含量测定的流动相为甲醇-0.1%磷酸水溶液(85∶15),检测波长254 nm;利用苯酚-硫酸法显色,运用UV测定总多糖含量,检测波长489 nm。结果:在同一时间取样,一体化方法与传统炮制方法对二苯乙烯苷含量均无显著性影响,随着蒸制时间的延长,何首乌传统炮制方法样品和一体化方法样品中二苯乙烯苷的质量分数呈逐渐降低的趋势;一体化方法与传统炮制方法对游离蒽醌类成分的含量具有显著性影响,且一体化方法中游离蒽醌类成分的含量明显低于传统炮制方法;2种炮制方法对总多糖含量均无显著性影响。结论:何首乌产地加工与炮制一体化方法制备的制何首乌质量较好、省时省力、可操作性强,具有一定的可行性。     

Src/NF-κB-targeted inhibition of LPS-induced macrophage activation and dextran sodium sulphate-induced colitis by Archidendron clypearia methanol extract

Ethnopharmacological relevance

Raw and processed Polygoni Multiflori Radix (PMR) are used in the prevention and treatment of non-alcoholic fatty liver disease (NAFLD), hyperlipidemia or related diseases. However, few researches compared the activities of raw and processed PMR on lipid metabolism regulation. Moreover, the active substances of Polygonum multiflorum are still not clearly elucidated.

Materials and methods

In this research, a sensitive, accurate and rapid in vitro model, steatosis hepatic L02 cell, was applied to compare the relative activities of raw and processed PMR on lipid metabolism regulation. Furthermore, the lipid regulation activities of emodin, physcion and 2,3,5,4′-tetrahydroxy-stilbene-2-O-β-d-glucoside (TSG) were evaluated. The steatosis L02 cells were obtained after cultured with 1% fat emulsion-10% fetal bovine serum (FBS)-RPMI 1640 medium for 48 h. Contents of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in L02 cells are evaluated after exposure.

Results

The intracellular TG contents were increased from 16.50 ± 1.29 mmol/L to 34.40 ± 1.36 mmol/L in steatosis L02 cells, while the intracellular contents of TC were increased from 5.07 ± 1.80 mmol/L to 11.79 ± 0.54 mmol/L. Water extract of raw PMR showed much remarkable TG-regulation and TC-regulation effects than its processed products. Emodin displayed the best TG regulation activity while TSG showed the best TC regulation activity. At the same time, the exposure of emodin and physcion could reduce the LDL-C contents in steatosis L02 cells.

Conclusions

On account of these in vitro results, raw PMR might have more satisfactory effects in clinic treatment of NAFLD or hyperlipidemia characterized by the elevation of cholesterol than processed PMR.     

In vitro effects of active components of Polygonum Multiflorum Radix on enzymes involved in the lipid metabolism

Ethnopharmacology relevance

Raw and processed Polygoni Multiflori Radix (PMR and PMRP) are used in the prevention and treatment of non-alcoholic fatty liver disease (NAFLD), hyperlipidemia or related diseases. In our previous research, 2, 3, 5, 4′-tetrahydroxy-stilbene-2-O-β-d-glucoside (TSG) displayed the most important role in the total cholesterol (TC) lowering effect among all the chemical constituents of Polygonum multiflorum. Emodin and physcion displayed more favorable triglyceride (TG) reducing effects than TSG. However, there are few researches focus on the approach and mechanism of how do Polygonum multiflorum exhibit good lipid regulation activity. The targeted sites of active substances of Polygonum multiflorum are still not clearly elucidated. This research pays close attention to how major chemical components of Polygonum multiflorum affect the TC and TG contents in liver cells.

Materials and methods

In this research, a sensitive, accurate and rapid in vitro model, steatosis hepatic L02 cell, was used to explore target sites of active chemical substances of Polygonum multiflorum for 48 h. Steatosis hepatic L02 cell was exposed to emodin, physcion and TSG, respectively. The contents of four key enzymes in the pathway of synthesis and decomposition of TC and TG were investigated after exposure. Meanwhile, the contents of lipid transfer protein were also tested. The diacylgycerol acyltransferase 1 (DGAT1) controlled the biosynthesis of TG from free fatty acids while 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMG-CoA reductase) limited the biosynthesis of TC. Hepatic triglyceride lipase (HTGL) and cholesterol 7α-hydroxylase (CYP7A) played the key role in the lipolysis procedure of TG and TC.

Results

The synthesis of TC and TG in steatosis L02 cells were apparently increased in the model group compared to the control group. Intracellular contents of HMG-CoA reductase and DGAT1 increased 32.33% and 56.52%, while contents of CYP7A and HTGL decreased 21.61% and 47.37%. Emodin, physcion and TSG all showed down-regulation effects on HMG-CoA reductase, while up-regulation effects on CYP7A. The most remarkable effect on HMG-CoA reductase was found on emodin. Emodin could reduce the DGAT1 content from 438.44±4.51 pg/mL in model group to 192.55±9.85 pg/mL (100 μm). The content of HTGL in 300 μm physcion group was 3.15±0.15 U/mL, which was more significantly effective than the control, lovastatin and fenofibrate group.

Conclusions

TSG could raise the content of CYP7A and then promote the lipolysis of cholesterol. Moreover, TSG also showed the best LDL-reducing effect. Emodin could inhibit HMG-CoA reductase and DGAT1, which were key enzymes in the synthesis of TC and TG. Physcion increased the content of HTGL, and then could boost the lipolysis of triglyceride. At the same time, physcion showed the best VLDL-reducing effect. In view of the above conclusions, we contributed the lipid regulation activity to an overall synergy of TSG, emodin and physcion.     

Orthogonal array design for optimizing extraction efficiency of active constituents from Jakyak-Gamcho Decoction, the complex formula of herbal medicines, Paeoniae Radix and Glycyrrhizae Radix

A complex formula composed of Paeonia lactiflora PALL. and Glycyrrhiza uralensis Fisch., which is called as Jakyak-Gamcho Decoction (JGD), has been used for a pain-relieving function and muscle spasms due to blood deficiency in the traditional medicine. In this study, the anti-inflammatory activity of JGD was evaluated based on the quantitative determinations and the relative proportions of six major constituents in the decoction mixture extracted by orthogonal array methods. Our results suggest that the three parameters are all crucial factors. The optimized conditions for extraction were therefore established [solvent (water); pH value (4); extraction number (4)]. We also optimized the extraction conditions related to anti-inflammatory activity [solvent (70% EtOH); pH value (6); extraction number (4)]. So, we found that the bioactivity was responsible for mixed components but not individual one. It was proportionally associated with the amounts of some components in the extracts of herbal medicines. When the proportion of the active components was similar to each other, they had the similar functions. Furthermore, the results could establish a model system for the quality assurance of herbal preparations, and provided a new paradigm of active components-pharmacodynamics, which is used for illustrating the connections between the bioactivities and the proportion of active constituents in the extracts of herbal medicines.