Variation and Cluster Analysis on Leaf Characters from Different Provenance Sources of Polygonum multiflorum Thunb

[Objective] The aim was to study the variation of leaf characters from different provenance sources of Polygonum multiflorum Thunb,as well as to carry out cluster analysis on P.multiflorum from different provenance sources to provide basis for the classification,identification,breeding and improved variety selection of P.multiflorum.[Method] Leaf shape characters of 31 copies of germplasm resources in the major distribution region of the whole country were determined,and the genetic variation of P.multiflorum leaves from different producing areas was analyzed.[Result] The leaf characters of single plant of the same experimental provenance source of P.multiflorum were relatively stable,the variation was mainly found on the single leaf area,1/2 leaf width,leaf width and other indicators;the variation of each leaf character among different provenance sources was obvious,and the variation was mainly found on the single leaf weight,leaf area,1/2 leaf width,leaf length and other indicators.The correlation analysis of each leaf character in P.multiflorum suggested that the single leaf area and single leaf weight showed extremely significant positive correlation with leaf length,1/2 leaf width,leaf width,leaf thickness and leaf stem length,while the single leaf area and single leaf weight showed significant negative correlation with WWR（leaf width/1/2 leaf width）and LWR（leaf length/1/2 leaf length）,in addition,several macroscopic leaf characters such as leaf length,1/2 leaf width,leaf width,leaf stem length showed extremely positive correlation.The main component analysis result suggested that the contribution rate of accumulation variance of the front three main components was up to 97.4%,which could better reflect the comprehensive performance of leaf characters of different provenance sources of P.multiflorum.The cluster analysis showed that the experimental 31 copies of P.multiflorum provenance sources should be divided into three classes,the first class was distributed in the Middle,Western of Guizhou,northwestern of Guangxi and western areas with higher altitude;the second class was distributed in Hunan,Hubei,Sichuan,Guangdong and the most area of Guangxi;the third class was distributed in Anhui,Jiangsu and Henan and Shandong.[Conclusion] Cluster analysis of leaf characters indicated that the kinds of provenance sources which the geographical position was closer could be got together.The study had provided a certain basis for the classification of P.multiflorum.

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.

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.

何首乌块根中砷、镉、汞和铅含量的检测及其富集特性

The contents of As, Cd, Hg and Pb in root tubers of annual and biennial plants of Polygonum multiflorum Thunb. in Jiangsu Province were 0.32, 0.24, 0, 1.25 and 0.20, 0.26, 1.67×10 -4, 1.32 mg·kg -1 respectively, which totally met the standards of GTSIEMPP (WM2—2001). Biology concentration factors (BCF) of Cd is 2.4-2.6, which indicates that the root tuber of P. multiflorum has a strong accumulative action to Cd. While the accumulative action to As, Hg and Pb is not strong, BCF are 0.025-0.015, 0-6.40×10 -4 and 0.06 respectively.

何首乌对D-半乳糖致衰大鼠的抗衰益智作用机制的研究

目的:探讨何首乌对D-半乳糖致衰大鼠海马的抗衰益智作用机制。方法:以D-半乳糖致衰大鼠为研究对象,采用Y-迷宫及一次被动回避反应,检测大鼠的学习记忆能力;用荧光标记测定海马突触体内游离钙离子浓度;用免疫组织化学结合图像分析测定海马内突触素(synaptophsin,P38)的灰度值。同时观察了何首乌对上述各项指标的影响。结果:老年模型组突触体内钙离子浓度显著升高,P38灰度值也明显高于青年对照组;何首乌给药组与老年模型组比较,学习记忆能力明显改善,突触体内钙离子浓度显著降低,P38灰度值也明显低于老年模型组。结论:何首乌可能通过抑制突触体内钙离子超载、提高P38含量起到抗衰益智作用。

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.

Simultaneous quantitative determination of eight active components in Polygonum multiflorum Thunb by RP-HPLC

A sensitive and specific reversed-phase high performance liquid chromatography （RP-HPLC） method was established for the simultaneous quantitative determination of eight active components, two stilbenes （resveratrol, polydatin） and six flavonoids （rutin, quercilrin, quercetin, luteotin, isoorientin, kaempferol）, in Polygonum multiflorum Thunb. The chromatographic separation was performed on a Kromasil C18 column （250 mm^4.6 mm, 5 ~tm）, and gradient elution was carried out with water-methanol as the mobile phase at a flow rate of 1 mL/min. Stilbenes and flavonoids were respectively detected at 320 nm and 350 nm with DAD. The correlation coefficients of all the calibration curves were found to be higher than 0.9995. The average recoveries were ranged from 96.8% to 102.5% with RSD less than 4.8% for these components. RP-HPLC was validated to be a robust method for the quantitative determination of active components in Polygonum multiflorum Thunb, and it could be used in the quality control of this traditional medicine.

Novel Stilbene Glycosides from Polygonum multiflorum

Two new stilbene glycosides (1 and 2), together with nine known compounds (3-11), were isolated from the water extract of Polygonum multiflorum Thunb. The structures of the new compounds were elucidated by their chemical properties and spectroscopic analyses, including extensive 2D NMR experiments. Compound 2 showed strong DNA cleavage activity, and compounds 1, 2 and 10 (2, 3, 4′, 5_tetrahydroxy_ trans _stilbene_2_O_β_ D _glucopyranoside) exhibited significant inhibition of lipid peroxidation.

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.

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.

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.

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.

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.

Transcriptome changes in Polygonum multiflorum Thunb. roots induced by methyl jasmonate

Transcriptome profiling has been widely used to analyze transcdptomic variation in plants subjected to abiotic or biotic stresses. Although gene expression changes induced by methyl jasmonate （MeJA） have been profiled in several plant species, no information is available on the MeJA-triggered transcriptome response of Polygonum multiflorum Thunb., a species with highly valuable medicinal properties. In this study, we used transcriptome profiling to investigate transcdptome changes in roots of P. mu/tiflorum seedlings subjected to a 0.25 mmol/L-MeJA rootirrigation treatment. A total of 18 677 differentially expressed genes （DEGs） were induced by MeJA treatment, of which 4535 were up-regulated and 14 142 were down-ragulated compared with controls. These DEGs were associated with 125 metabolic pathways. In addition to various common primary and secondary metabolic pathways, several sec- ondary metabolic pathways related to components with significant pharmacological effects were enriched by MeJA, including arachidonic acid metabolism, linoleic acid metabolism, and stilbenoid biosynthesis. The MeJA-induced transcdptome changes uncovered in this study provide a solid foundation for future study of functional genes controlling effective components in secondary metabolic pathways of P. multiflorum.

补肾健骨中药防治骨质疏松机制研究概况

补肾健骨中药治疗骨质疏松(OP)疗效确切,可作为主要治疗方法。(1)促进成骨细胞增殖、ALP分泌和分化及相关基因表达,抑制破骨。(2)调节雌激素,杜仲可提高体内雌激素(E2)水平及IGF-1含量,淫羊藿有类植物雌激素样作用,当归可促进β-EP合成,起到类雌激素样作用。(3)抑制MSCs成脂与促进MSCs成骨过程,杜仲调控Wnt信号转导途径增进BMSCs分化成骨过程,抑制与成脂过程有关转录因子结合蛋白表达;淫羊藿抑制骨髓干细胞成脂过程,减弱C/EBPα、成脂转录因子PPARγ基因表达。(4)调控OPG/RANK/RANKL系统,杜仲增进MC3T3-E1型成骨细胞增殖分化,淫羊藿上调OPGmRNA表达抑制OPGLmRNA表达。(5)调控骨骼钙磷代谢及氧化应激,何首乌提高肾脏1α-羟化酶理化活性,调节钙磷代谢;何首乌有明显抗氧化作用,仙茅可提高抗氧化应激能力,淫羊藿可对抗缺氧、清除活性氧作用。(6)未来应全面分析补肾健骨类中药复方制剂,结合现代实验检测,详细探讨补肾中药防治OP机制。

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.

壳聚糖和中药复方对实验性大鼠脂肪肝的防治作用

目的:观察壳聚糖和中药首乌、丹参、牛膝的复方对大鼠脂肪肝的预防作用。方法:在小剂量的四氯化碳致肝损伤的基础上,合并应用高脂饮食复制大鼠脂肪肝模型,同时分别给予不同浓度的药物,以肝脏的脂质含量为衡量指标,观察药物预防结果。结果:四氯化碳损伤肝脏合并高脂饮食可引起大鼠肝脏明显的脂肪变性,表现为肝组织甘油三酯(TG)、总胆固醇(TC)含量显著增加,SOD活力明显降低,镜检显示肝细胞肿胀,内部充满大小不等的脂滴,而药物防治组大鼠与模型组相比肝组织TG、TC值显著降低,肝细胞的脂变程度明显小于模型组,表现为脂滴减少,脂变面积缩小,SOD活力逐渐升高并恢复正常,其改善程度随用药剂量的增加而增加,应用中、高剂量的药物还可使大鼠血清丙氨酸转氨酶活性明显降低。结论:药物对脂肪肝的形成具有明显的预防作用,其机理除了减少脂肪自肠道的吸收外,还可能有保护肝脏、提高肝抗氧化能力,促进其对脂质的氧化与转运功能的作用。

何首乌中二苯乙烯苷的醇提工艺

探索了从何首乌中提取二苯乙烯苷的工艺流程。在比较不同提取方法的基础上 ,确定选用乙醇为提取溶剂。通过单因素和中心组合实验 ,对影响何首乌中二苯乙烯苷提取的因素进行了探讨和研究 ,运用响应面分析法确定了提取工艺的最优化条件 :料液比 (g∶mL)为 1∶9,提取时间为66min ,乙醇体积分数为 61 1 % ,提取次数为 2次。在此条件下 ,二苯乙烯苷的得率可达 6 3 9%。

黑豆汁制首乌的质量控制研究

目的 为提高制首乌的质量标准提供参考依据。方法 采用薄层色谱法 ,建立黑豆汁制首乌的鉴别方法。结果 该法操作简便 ,专属性及重现性好。

何首乌脑保护作用机理研究的进展

本文就近10年何首乌及其提取物有效成分二苯乙烯苷(2,3,5,4’-四羟基二苯乙烯-β-D-葡萄糖苷)对脑保护作用的机理研究进行了综述。它们的主要作用如下:(1)钙通道拮抗剂的作用;(2)抗氧化剂的作用;(3)拟胆碱药和胆碱脂酶抑制剂的作用;(4)调节细胞凋亡;(5)延缓衰老。何首乌的脑保护作用机理是多靶点、多环节、多途径的。因此,何首乌在老年痴呆、帕金森氏病、血管性痴呆等老年神经系统疾病的防治中具有很高的应用价值。