In vivo recognition of bioactive substances of Polygonum multiflorum for protecting mitochondria against metabolic dysfunction-associated fatty liver disease

BACKGROUND Metabolic dysfunction-associated fatty liver disease(MAFLD)is a severe threat to human health.Polygonum multiflorum(PM)has been proven to remedy mitochondria and relieve MAFLD,but the main pharmacodynamic ingredients for mitigating MAFLD remain unclear.AIM To research the active ingredients of PM adjusting mitochondria to relieve highfat diet(HFD)-induced MAFLD in rats.METHODS Fat emulsion-induced L02 adipocyte model and HFD-induced MAFLD rat model were used to investigate the anti-MAFLD ability of PM and explore their action mechanisms.The adipocyte model was also applied to evaluate the activities of PM-derived constituents in liver mitochondria from HFD-fed rats(mitochondrial pharmacology).PM-derived constituents in liver mitochondria were confirmed by ultra-high-performance liquid chromatography/mass spectrometry(mitochondrial pharmacochemistry).The abilities of PM-derived monomer and monomer groups were evaluated by the adipocyte model and MAFLD mouse model,respectively.RESULTS PM repaired mitochondrial ultrastructure and prevented oxidative stress and energy production disorder of liver mitochondria to mitigate fat emulsion-induced cellular steatosis and HFD-induced MAFLD.PM-derived constituents that entered the liver mitochondria inhibited oxidative stress damage and improved energy production against cellular steatosis.Eight chemicals were found in the liver mitochondria of PM-administrated rats.The anti-steatosis ability of one monomer and the anti-MAFLD activity of the monomer group were validated.CONCLUSION PM restored mitochondrial structure and function and alleviated MAFLD,which may be associated with the remedy of oxidative stress and energy production.The identified eight chemicals may be the main bioactive ingredients in PM that adjusted mitochondria to prevent MAFLD.Thus,PM provides a new approach to prevent MAFLD-related mitochondrial dysfunction.Mitochondrial pharmacology and pharmacochemistry further showed efficient strategies for determining the bioactive ingredients of Chinese medicines that adjust mitochondria to prevent diseases.

Two new coumarin glucosides biosynthesized by transgenic hairy roots of Polygonum multiflorum

The glycosylation of hydroxylcoumarin was investigated by using suspension cultures of hairy roots of Polygonum multiflorum. Two new coumarin glucosides （3 and 4） were biosysthesized by regioselectively glycosylation of corresponding substrates （1 and 2） in the system. The structures of two products were identified as 7-hydroxy-4-methylcoumarin 5-O-β-D-glucopyranoside and 7- hydroxy-3,4-dimethylcoumarin 5-O-β-D-glucopyranoside on the ground of chemical and spectroscopic methods, respectively. 2007 Rong Min Yu. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Concurrent severe hepatotoxicity and agranulocytosis induced by Polygonum multiflorum:A case report

BACKGROUND Various types of drug-induced liver injury are induced by Polygonum multiflorum(PM);however,it rarely causes neutropenia.Herein,we report the case of a 65-year-old woman with concurrent severe hepatotoxicity and agranulocytosis induced by PM.CASE SUMMARY A 65-year-old woman reported with severe hepatotoxicity and agranulocytosis 17 d after ingestion of PM.The results of the Roussel Uclaf Causality Assessment Method demonstrated a highly probable relationship between hepatotoxicity and PM,with a total score of 10.The Naranjo algorithm results indicated that agranulocytosis had a probable relationship with PM,with an overall score of 6.Granulocyte colony-stimulating factor(for once),a steroid,compound glycyrrhizin,and polyene phosphatidylcholine therapy were initiated.After 15 d of treatment,there was a gradual improvement in liver biochemistry,leukocytes,and neutrophils levels.CONCLUSION Concurrent hepatotoxicity and agranulocytosis are rare and critical adverse drug reactions of PM,which should be highly valued.

Hypoglycemic effects of naturally processed Polygonum multiflorum extract in KK CgAy/J mice and its mechanism of action

In 2021,there are approximately 537 million adults ageing 20-79 years affected by diabetes worldwide and the number is rising rapidly,hence it is important to manage and control diabetes mellitus and its associated complications.Food is one of the key factors in preventing and combating diseases such as diabetes.Both as a food and an herbal medicine,Polygonum multiflorum(PM)has been used as an anti-aging tonic,for hair darkening in traditional Chinese medicine for several centuries.The recent research effort of PM has been focused on antioxidant,anti-ageing and anti-tumor properties.In the present study,we utilized the traditional processing of harvested raw PM,and identified several stilbene components and then evaluated the potential anti-diabetic effects of the processed PM extract(PME).PME(0.075%)was given to diabetic mice(KK CgAy/J)in drinking water and after 7 weeks,PME-treated mice had significantly lower glucose levels than mice in the diabetic control group(P<0.01).The mechanism was explored with ELISA and Western blotting and results suggested that the effect was through maintainingβ-cell function.

N-Acetyl Cysteine to Treat <i>Polygonum multiflorum</i>Hepatotoxicity: A Case Report

Background: Polygonum multiflorum (brand name Fo-ti) is a Chinese herb used as an antioxidant, antiaging agent, and liver and kidney health. It can cause acute hepatotoxicity and even death. There is no published data on the effectiveness of N-Acetyl cysteine (NAC) as a sole effective therapy. Case: A 23-year-old, healthy male, presented with jaundice and malaise of one-month duration. He had been taking a Chinese herb named Fo-Ti for 2 months. Physical examination revealed jaundice, with no stigmata of chronic liver disease. His laboratory results showed transaminitis with markedly elevated transaminases. Testings for infectious and autoimmune causes were unremarkable. The patient was admitted to the hospital. Fo-Ti was stopped and he was started on oral NAC as peroral protocol. He was discharged home on esomeprazole and oral NAC. He returned on the second day with worsening jaundice. He was readmitted and esomeprazole was discontinued. NAC was continued and his liver enzymes eventually improved. The supplement was tested by gas-chromatography mass spectrometry and measured the presence of emodin. Discussion: This case highlights the potential benefit of NAC as treatment of Polygonum multiflorum induced hepatotoxicity. The proposed mechanism of emodin induced liver injury is through disturbing glutathione metabolism in liver cells. This could explain the favorable response as NAC has the ability to replenish intracellular glutathione. Conclusion: Polygonum multiflorum use has been associated with severe and prolonged hepatotoxicity. Oral NAC should be considered in the treatment of these patients due to the favorable risk-benefit ratio.

Single-nucleotide polymorphisms of HLA and Polygonum multiflorum-induced liver injury in the Han Chinese population

BACKGROUND Polygonum multiflorum is one of the leading causes of herb-induced liver injury in China.HLA-B*35:01 is reported to be a potential biomarker of Polygonum multiflorum-induced liver injury(PM-DILI).However,little is known about the relationship between single-nucleotide polymorphisms(SNPs) and PM-DILI.AIM To identify SNPs that indicate susceptibility to PM-DILI METHODS We conducted a systematic study enrolling 382 participants from four independent hospitals,including 73 PM-DILI patients,118 patients with other drug-induced liver injury(other-DILI) and 191 healthy controls.Whole-exome sequencing was performed for 8 PM-DILI patients and 8 healthy controls who were randomly selected from the above subjects.Nineteen SNPs that showed high frequencies in the 8 PM-DILI patients were selected as candidate SNPs and then screened in 65 PM-DILI patients,118 other-DILI patients and 183 healthy controls using the MassARRAY system.HLA-B high-resolution genotyping was performed for the 73 PM-DILI and 118 other-DILI patients.The Han-MHC database was selected as a population control for HLA-B analysis.P <6.25 x 103 after Bolferroni correction was considered significant.RESULTS The frequencies of rslll686806 in the HLA-A gene,rs1055348 in the HLA-B gene,and rs202047044 in the HLA-DRB1 gene were significantly higher in the PM-DILI group than in the control group [27.2% vs 11.6%,P=1.72×105,odds ratio(OR)=3.96,95% confidence interval(Cl):2.21-7.14;42.5% vs 8.6%,P=1.72×10-19 OR=13.62,95% CI:7.16-25.9;22.9% vs 8.1%,P=4.64×106,OR=4.1,95% CI:2.25-7.47].Only rs1055348 showed a significantly higher frequency in the PM-DILI group than in the other-DILI group(42.5% vs 13.6%,P=1.84×10-10,OR=10.06,95% Cl:5.06-20.0),which suggested that it is a specific risk factor for PM-DILI.rs1055348 may become a tag for HLA-B*35:01 with 100% sensitivity and 97.7% specificity in the PM-DILI group and 100% sensitivity and 98.1% specificity in the other-DILI group.Furthermore,HLA-B*35:01 was confirmed to be associated with PM-DILI with a frequency of 41.1% in the PM-DILI group compared with 11.9%(P=4.30×10-11,OR=11.11,95% CI:5.57-22.19) in the other-DILI group and 2.7%(P=6.22×10-166,OR=62.62,95% Cl:35.91-109.20) in the Han-MHC database.CONCLUSION rslll686806,rs1055348,and rs202047044 are associated with PM-DILI,of which,rs1055348 is specific to PM-DILI.As a tag for HLA-B*35:01,rs1055348 may become an alternative predictive biomarker of PM-DILI.

Optimization of Preparation and Brewing Process of Radix Polygonum Multiflorum Preparata-Herba Taxilli Tea Bag by Orthogonal Test Combined with Comprehensive Weighted Score Analysis

[Objectives]This study aimed to optimize the preparation process of Radix Polygonum Multiflorum Preparata-Herba Taxilli tea bags.[Methods]Taking the comprehensive weighed score of total flavonoid content,soup color,aroma and taste as an indicator,the preparation process of tea bags was optimized using an orthogonal test.[Results]The results of the orthogonal test show that the optimal preparation process for the Radix Polygonum Multiflorum Preparata-Herba Taxilli tea bags are as follows:particle size of 14 meshes,solid to liquid ratio of 1∶40,brewing temperature of 70℃,and brewing time of 10 min.[Conclusions]The optimized preparation process is reasonable and feasible.It is suitable for the production of Radix Polygonum Multiflorum Preparata-Herba Taxilli tea bags.

Transcriptional regulation effect of constituents in Polygonum multiflorum Thunb. based on human progesterone X receptor(hPXR) mediated CYP3A4 rapid screening system

OBJECTIVE This study investigated transcriptional regulation of the main chemical con.stituents of Polygonum multiflorum Thunb.including Stilbene Glucoside(THSG) and anthraquinone constituents(Emodin,Rhein,Aloeemodin,Chrysophanol and Physcion) and six potential liver injury constituents(gallic acid,quercetin,luteolin,kaempferol,resveratrol) on mediated by PXR CYP3A4.Early establishment of pregnane X receptor mediated CYP3A4 drug induced rapid screening technique was used to determine the effects of these constituents.METHODS First,effect of constituents on the cell activity was detected by MTS cell viability assay.IC50 was calculated.Second,the expression vector and reporter vector were co-transfected into Hep G2 cells,10 μmol·L^(-1) Rifampicin as a positive control,10 μmol·L^(-1) Ketoconazole as a negative control.After treated with different concentrations of(the an.thraquinone constituents concentrations were 2.5,5 and 10 μmol·L^(-1);the concentrations of Gallic Acid,Quercetin,Luteolin,Kaempferol,Apigenin,Resveratrol concentrations were 5,10 and 20 μmol·L^(-1)) for24 h,the cells were tested for dual luciferase activity.RESULTS The results show that the inhibitory ef.fect of THSG,Chrysophanol,Emodin,Rhein and Aloeemodin on CYP3A4 was inhibited by co-transfec.tion of pcDNA3.1 and pGL4.17-CYP3A4.The expression of pcDNA3.14-PXR and pGL4.17-CYP3A4 was induced by the four constituents.Besides,Emodin has a directly inducing effect.Four anthraqui.none constituentscan induce the effect of CYP3A4 by PXR,but Emodin can directly induce CYP3A4.THSG can inhibit CYP3A4,but in the presence of PXR plasmid can induce CYP3A4.For the six poten.tial liver injury constituents,results show that the plasmid pcDNA3.1 was cotransfected with pGL4.17-CYP3A4 regulation of Gallic Acid and Resveratrol on CYP3A4 inhibitory effects of Quercetin,Luteolin,Kaempferol have an induce effect;after pcDNA3.14-PXR and pGL4.17-CYP3A4 cotransfected,Quercetin,Luteolin,Kaempferol,Apigenin,Resveratrol have induced effect,three constituents' induc.tion effect had significant difference.CONCLUSION 12 kinds of Polygonum multiflorum Thunb.constit.uents have inhibitory or activating effects on CYP3A4,after the participation of PXR,9 components have induced effects on CYP3A4,and the induction effect of 6 components has significant difference.The results suggested that we should pay attention to potential drug interactions when combined with Polygonum multiflorum Thunb.,and improve safety and efficacy.

Evidence chain-based causality identification in herb-induced liver injury： exemplification of a well-known liver-restorative herb Polygonum multiflorum

Herbal medicines have recently been recognized as the second most common cause of drug-induced liver injury （DILI） in the United States. However, reliable methods to identify the DILl causality of some herbs, such as Heshouwu （dried root ofPolygonum multiflorum）, remain lacking. In this study, a total of 12 307 inpatients with liver dysfunction and 147 literature-reported cases of Heshouwu DILI were screened. A general algorithm indicated that only 22.5% （9/40） and 30.6% （45/147） of all hospitalization and literature case reports, respectively, demonstrate the high probability of DILI causality of Heshouwu. By contrast, 95% （19/20） of all cases prospectively investigated by pharmacognosy, phytochemistry, and metabolomic tests exhibited highly probable causality, including a patient who was previously incorrectly attributed and a case that was excluded from Heshouwu causality by pharmacognostic evidence. Toxin （heavy metals, pesticides, and mycotoxins） contamination was also excluded from Heshouwu DILI causality. The objectivity of these screening methods for Heshouwu DILI diagnosis addresses safety concerns regarding stilbene-containing herbal medicines and dietary supplements.

Screening for main components associated with the idiosyncratic hepatotoxicity of a tonic herb, Polygonum multiflorum

The main constituents of a typical medicinal herb, Polygonum multiflorum （Heshouwu in Chinese）, that induces idiosyncratic liver injury remain unclear. Our previous work has shown that cotreatment with a nontoxic dose of lipopolysaccharide （LPS） and therapeutic dose of Heshouwu can induce liver injury in rats, whereas the solo treatment cannot induce observable injury. In the present work, using the constituent ＂knock-out＂ and ＂knock-in＂ strategy, we found that the ethyl acetate （EA） extract of Heshouwu displayed comparable idiosyncratic hepatotoxicity to the whole extract in LPS-treated rats. Results indicated a significant elevation of plasma alanine aminotransferase, aspartate aminotransferase, and liver histologic changes, whereas other separated fractions failed to induce liver injury. The mixture of EA extract with other separated fractions induced comparable idiosyncratic hepatotoxicity to the whole extract in LPS-treated rats. Chemical analysis further revealed that 2,3,5,4＇-tetrahydroxy trans-stilbene-2-O-p-glucoside （trans-SG） and its cis-isomer were the two major compounds in EA extract. Furthermore, the isolated cis-, and not its trans-isomer, displayed comparable idiosyncratic hepatotoxicity to EA extract in LPS-treated rats. Higher contents of cis-SG were detected in Heshouwu liquor or preparations from actual liver intoxication patients associated with Heshouwu compared with general collected samples. In addition, plasma metabolomics analysis showed that cis-SG-disturbing enriched pathways remarkably differed from trans-SG ones in LPS-treated rats. All these results suggested that cis-SG was closely associated with the idiosyncratic hepatotoxicity of Heshouwu. Considering that the cis-trans isomerization oftrans- SG was mediated by ultraviolet light or sunlight, our findings serve as reference for controlling photoisomerization in drug discovery and for the clinical use of Heshouwu and stilbene-related medications.

New phenolic constituents obtained from Polygonum multiflorum

Objective:To isolate the phenolic compounds obtained from the dried roots ofPolygonum multiflorum and investigate their pharmacological activities.Methods:The chemical constituents were isolated and purified by combining them with a macroporous resin(DM-8),MCI gel,and Sephadex LH-20 and by performing ODS column chromatography.Their structures were elucidated by 1D and 2D NMR analyses,as well as mass spectrometry.The isolated compounds were evaluated to determine their hepatoprotective andα-glucosidase inhibitory activities in vitro.Results:Two phenolic compounds,namely,polygonimitin E(1)and polygonimitin F(2),were isolated from the dried roots of P.multiflorum.Compound 2(10μmol/L)only showed moderate hepatoprotective activity against N-acetyl-p-aminophenol(APAP)-induced HepG2 cell damage.Unfortunately,these two compounds exhibited noα-glucosidase inhibitory activity.Conclusion:Compounds 1 and 2 were new compounds.Compound 2 could be one of the potential hepatoprotective constituents of P.multiflorum.

Biotransformation of 4-Hydroxybenzen Derivatives by Hairy Root Cultures of Polygonum multiflorum Thunb

The biotransformation of four 4-hydroxybenzen derivatives （1,4-benzenediol （compound 1）, 4-hydroxybenzaldehyde （compound 2）, 4-hydroxybenzyl alcohol （compound 3） and 4-hydroxybenzoic acid （compound 4）） by the hairy root cultures of Polygonum multiflorum Thunb. as a new biocatalyst was investigated. It was found that the substrates were transformed to their corresponding glucosides, 4-hydroxyphenyl β-D-glucopyranoside （arbutin, compound la）, 4-hydroxymethylphenyl β-D-glucopyranoside （gastrodin, compounds 2a, 3a） and 4-carboxyphenyl α-D- glucopyranoside （compound 4a）, respectively. In the meantime, the hairy roots of P. multiflorum were able to stereoselectively and regioselectively glucosylate phenolic hydroxyl groups of compounds 1-4, but the cultures could not glucosylate the aldehyde group of compound 2 or the benzyllc hydroxyl group of compound 3, and no glucosyl esterification of carboxyl groups of compound 4 was detected. On the other hand, the result also showed that the hairy roots of P. multiflorum were able to reduce the 4-hydroxybenzaldehyde to its corresponding alcohol. This is the first report that substrate 4 has been converted into its α-D-glucopyranoside by a plant biotransformα- tion system.

Idiosyncratic Drug-Induced Liver Injury Linked to Polygonum multiflorum:A Case Study by Pharmacognosy

The use of Chinese medicinal herbs（CMH）continues to expand rapidly across the world with a deep belief that herbs are safe because they are＂natural＂.（1）However,a survey of the literature indicates that CMH related hepatotoxicity is increasing and has been a worldwide problem.（2,3）Polygonum multiflorum Thunb.（P.multiflorum）,officially listed in the Pharmacopoeia of the People＇s Republic of China.

何首乌叶不同萃取物的抗炎活性及其化学成分鉴定

目的:研究何首乌叶的成分,为其功能食品的研发提供基础数据。方法:对何首乌叶醇提取物进行有机溶剂萃取,分别得到何首乌叶石油醚、二氯甲烷、乙酸乙酯、正丁醇、水等不同萃取物,并对不同萃取物进行抗炎活性检测,后续通过硅胶柱色谱、C18柱色谱结合半制备液相等方法对抗炎活性较好的成分进行分离、纯化以及结构鉴定,结果:在质量浓度0~100μg/mL范围内,乙酸乙酯萃取物对RAW264.7细胞存活率无明显的抑制作用,且剂量依赖地抑制NO的生成。石油醚和二氯甲烷萃取物在质量浓度0~25μg/mL时,均无明显细胞毒性,并在25μg/mL时,二氯甲烷萃取物对NO的抑制率(51.47%)高于石油醚萃取物的抑制率(43.91%)。从二氯甲烷和乙酸乙酯萃取物中分离、鉴定出11个化合物,分别为2,3,4,6-三羟基苯乙酮-3-O-β-D-glucoside(1)、杨梅苷(2)、槲皮苷(3)、阿福豆苷(4)、β-谷甾醇(5)、正十六烷-5,8,11三烯酸(6)、二十六烷(7)、α-亚麻酸(8)、山柰酚-3-O-芸香糖苷(9)、肉豆蔻酸(10)和槲皮素(11)。结论:何首乌叶具有很好的抗炎活性,化合物2,4,6,7,8,9,10,11均为首次从何首乌中分离得到。

何首乌肝损伤机制及减毒方法研究进展

何首乌Polygonum multiforum Thunb.是临床常用滋补中药,同时作为一种保健中药在药膳和食膳中广泛应用,但是近年来由于不时出现何首乌及其制品致肝损伤的报道,引起了研究者的警觉及关注。本文通过整理分析关于何首乌致肝损伤的国内外临床相关报道,从何首乌致肝毒性的物质基础二苯乙烯苷类、蒽醌类化合物和没食子酸等成分及其潜在肝毒性机制(包括代谢酶改变、氧化应激、炎症反应、线粒体损伤、肝细胞异常凋亡等方面)进行归纳总结,探讨炮制减毒和配伍减毒两种减毒方法对何首乌肝毒性的影响和机制。

何首乌属化学成分与药理作用研究进展

何首乌属植物作为一类重要的药用植物资源,一直以来都是国内外学者研究的热点。何首乌属植物化学成分的研究表明,该属植物中主要含有二苯乙烯类、黄酮类、醌类、生物碱类等多种化学成分。民间用药经验及现代研究证实何首乌属植物具有抗肿瘤、抗炎、抗菌、调节免疫等药理活性。现对何首乌属植物近年来化学成分和药理作用的研究进行总结归纳,旨在为何首乌属植物进一步的研究和合理开发利用提供参考。

药对何首乌-牛膝抗骨质疏松的作用机制研究

目的:利用网络药理学研究牛膝何首乌联用抗骨质疏松的作用机制。方法:运用中药系统药理学数据库与分析平台(TCMSP)与文献检索筛选何首乌、牛膝两味中药所含的活性成分。利用Disgent数据库搜索与骨质疏松相关的蛋白靶点;利用Pubchem平台,预测何首乌、牛膝所含活性成分所作用的人类蛋白靶点。分析牛膝何首乌药的交集靶点,并将交集靶点进行蛋白网络分析,构建药物-成分-靶点的网络图。通过微生信平台对关键基因作用的靶向细胞进行富集分析。结果:确定何首乌成分5个,牛膝成分10个,二者共有成分1个,获得药对与疾病交集基因35个;PPI蛋白分析获得10个主要作用成分;绘制GO分析可视图、KEGG分析可视图并对其进行解析,解析结果表明何首乌牛膝主要成分通过抑制骨吸收,诱导骨形成等通路来减轻骨质疏松症的发展。结论:牛膝何首乌药对抗骨质疏松的作用机制可能是通过靶点与共有通路完成的,并且该药对可通过多通路、多靶点、多成分、多层次发挥对骨质疏松症的治疗作用。

基于网络药理学探究何首乌有效成分含量与抗肿瘤活性

目的基于网络药理学探究广东省不同产地的12批次何首乌主要活性成分EBYXG和蒽醌类化合物(大黄素、大黄素甲醚)的抗氧化和抗肿瘤活性机制,并通过测定化合物质量分数,对何首乌的质量进行评价。方法通过HPLC法测定12批次何首乌样品中EBYXG与大黄素、大黄素甲醚的质量分数。通过SwissTargetPrediction、SuperPred数据库筛选成分作用靶点,利用Uniprot进行校准。通过String数据库、Cytoscape3.7.2软件筛选核心靶点,绘制蛋白互作网络;通过DAVID数据库进行GO和KEGG富集分析预测药物作用机制。利用AutoDock平台进行分子对接,预测药物作用靶点程度,结合细胞及抗氧化实验,验证网络药理学分析结果。结果12批何首乌样品中EBYXG、蒽醌类化合物含量分别在1.526%~3.936%、0.129%~1.390%。找到“成分-疾病”交集基因共179个。GO与KEGG富集分析结果显示EBYXG、大黄素、大黄素甲醚主要通过PI3K-Akt、HIF-1、MAPK、cAMP等信号通路发挥抗氧化、抗肿瘤作用。分子对接结果显示,活性成分可以与靶点蛋白中心相结合。抗氧化及细胞试验证明,12批次何首乌均具有良好的抗氧化、抗肿瘤活性。结论广东省不同产地何首乌中的主要有效成分EBYXG、大黄素、大黄素甲醚质量分数均达到2020年版《中华人民共和国药典》规定。基于网络药理学及药理试验证实何首乌可通过多成分、多靶点、多信号通路产生抗氧化、抗肿瘤的功效,为何首乌在抗肿瘤领域的应用提供参考。

不同干燥方式的九制何首乌对D-半乳糖致衰老小鼠的抗衰老作用

目的:探究不同干燥方式的九制何首乌对D-半乳糖诱导衰老小鼠的影响。方法:将50只ICR小鼠随机分成空白组、模型组、阳性对照组、九蒸九晒制首乌组、九蒸九烘制首乌组,每日腹腔注射D-半乳糖(0.5 g·kg^(-1))构建小鼠衰老模型,造模同时给药,持续12周。应用Morris水迷宫实验方法测定小鼠的空间记忆和学习水平,计算各组小鼠胸腺、脾脏指数,检测小鼠血清及脑、肝、肾匀浆中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)水平,检测小鼠肝肾功能水平。结果:与空白对照组相比,模型组空间记忆和学习能力明显降低,体质量、脏器指数明显下降,血清以及全脑、肝、肾匀浆中SOD、GSH-Px水平明显下降,而MDA水平显著上升,血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶、碱性磷酸酶、肌酐、尿素水平显著性上升;药物干预组与模型组相比,空间记忆和记忆水平明显改善,体质量、脏器系数显著增加,改善D-半乳糖诱导的氧化应激,提高SOD、GSH-Px水平,抑制MDA含量的升高,肝肾功能趋于正常。两种干燥方式的给药组小鼠的指标相当,无明显差异。结论:不同干燥方法的九制何首乌的水煎液对D-半乳糖致衰老小鼠具有抗衰老作用,且两种方法的抗衰老作用无显著差异。

HPLC-QAMS法同时测定何首乌防脱育发精华液中9种成分含量

建立高效液相色谱一测多评法(HPLC-QAMS法)同时检测何首乌防脱育发精华液中9种成分的含量,并验证所建立方法的可行性和准确性。以槲皮苷为含量参照物,甲醇-0.1%磷酸为流动相,分别采用外标法和HPLC-QAMS法同时测定12批何首乌防脱育发精华液中大黄素、大黄素甲醚、杨梅苷、槲皮苷、杨梅素、槲皮素、山柰酚、穗花杉双黄酮、扁柏双黄酮的含量,对比两种方法的结果差异,验证HPLC-QAMS法的适用性。结果显示,9种成分各线性范围良好,相关系数均大于0.999,检出限为0.027~0.151μg/mL,定量限为0.076~0.449μg/mL。以槲皮苷为内参比物质,大黄素、大黄素甲醚、杨梅苷、杨梅素、槲皮素、山柰酚、穗花杉双黄酮、扁柏双黄酮的相对校正因子分别为0.7537,0.8369,1.0192,2.9913,1.6875,2.2369,1.0963和1.2493。两种方法测得的何首乌防脱育发精华液中9种成分的含量差异均无统计学意义(P>0.05)。以槲皮苷为含量参照物建立的相对校正因子重现性好,HPLC-QAMS法可用于何首乌防脱育发精华液中9种成分的质量评价。