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.

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.

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.

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.

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.

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.

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.

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.

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.

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。

肥水处理对一年生何首乌块根产量的影响

为探明一年生何首乌生长的水肥田间管理方法,以大棚盆栽何首乌为试材,用氮、磷、钾、水4因素3水平正交试验设计L^(9)(3^(4))方法,测定一年生何首乌产量及最大块根重量。结果表明,氮、磷、钾、水4个因素对何首乌块根产量的影响,其主次顺序依次为土壤持水量、磷肥、氮肥和钾肥;水和磷两个因素对何首乌产量的影响达到了极显著水平,而氮与钾两个因素未达到显著水平;产量随土壤持水量的增大呈先上升后下降的趋势,最大产量出现在水因素为W_(2)时,为297.86 g/株;各处理何首乌最大块根重量在24.57~80.95 g的范围内变化;何首乌最大块根重量与其产量呈极显著正相关。一年生何首乌产量适宜土壤水分含量为65%田间持水量,磷肥单株每盆需要P_(2)O_(5)2.16 g。

何首乌提取物对注意缺陷多动障碍模型大鼠BDNF/TrkB及其下游信号通路的影响

目的观察何首乌提取物对注意缺陷多动障碍(attention deficit hyperactivity disorder,ADHD)模型大鼠的影响并探讨其作用机制。方法选择自发性高血压大鼠(spontaneously hypertensive rat,SHR)为ADHD模型,随机分为模型组、托莫西汀组(4.5 mg/kg),何首乌低(0.54 g/kg)、中(1.08 g/kg)、高(2.16 g/kg)剂量组,另设Wistar京都大鼠为正常组,每组10只。各组大鼠每日给予相应药物灌胃,4周后观察各组大鼠行为学,ELISA法检测前额叶皮质中二十二碳六烯酸(docosahexaenoic acid,DHA)含量,Western blot和RT-qPCR法检测前额叶皮质和海马组织中脑源性神经营养因子(brain derived neurotrophic factor,BDNF)/酪氨酸蛋白激酶受体B(tyrosine kinase receptor B,TrkB)及其下游信号通路中生长因子受体结合蛋白2(growth factor receptor-bound protein 2,Grb2)、肌肉RAS癌基因(muscle RAS oncogene,Ras)3的表达水平。结果托莫西汀和何首乌均能有效控制SHR大鼠多动、冲动行为。与正常组比较,模型组大鼠前额叶皮质中DHA含量降低(P<0.05),BDNF、TrkB、Grb2、Ras3蛋白和mRNA表达明显降低(P<0.01),海马中BDNF、TrkB、Grb2、Ras3蛋白和BDNF、Ras3 mRNA表达明显下调(P<0.05)。给药4周后,在前额叶皮质中,何首乌低、中、高剂量组大鼠DHA含量均显著升高(P<0.01),BDNF、TrkB、Grb2、Ras3的mRNA表达水平均显著上升(P<0.01),何首乌低、中剂量组大鼠BDNF、TrkB、Grb2蛋白,高剂量组大鼠BDNF、Grb2、Ras3蛋白表达水平均显著上升(P<0.05);在海马中,何首乌高剂量组大鼠BDNF、TrkB、Grb2、Ras3蛋白和mRNA表达均明显增加(P<0.05)。结论何首乌提取物能够明显改善SHR多动、冲动行为,其机制可能与调节BDNF/TrkB及其下游信号通路有关。

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。

酵母发酵何首乌醇提物对小鼠肝脏和免疫指标的影响

试验旨在研究用酵母发酵的何首乌醇提取物对小鼠肝脏和免疫细胞的影响。选取60只昆明雄鼠随机分为4组,低剂量组、中剂量组、高剂量组分别灌服相当于生药剂量11、21、42 g/kg的发酵何首乌醇提物溶液,对照组灌服双蒸水,连续灌胃30 d。结果显示,给药期间肉眼观察小鼠无明显毒性表现,体重稳定上升,脏器系数均无显著差异(P>0.05);高剂量组谷草转氨酶(AST)和谷丙转氨酶(ALT)活性明显增高,显著高于其他组(P<0.05);高剂量组肿瘤坏死因子-α(TNF-α)水平升高,显著高于对照组(P<0.05),白介素-17(IL-17)、Bcl2关联X蛋白(Bax)、B淋巴细胞瘤-2基因(Bcl-2)各组间差异不显著(P>0.05);H.E.染色结果显示,高剂量组出现少量炎性浸润。各组小鼠脾脏中的T淋巴细胞(CD3^(+)、CD4^(+)、CD8^(+))、B淋巴细胞(CD19^(+))百分比均差异不显著(P>0.05)。说明酵母发酵何首乌醇提物在低、中剂量时对小鼠无明显毒性,在高剂量时出现了轻微的炎症现象;酵母发酵的何首乌对小鼠脾脏内的免疫细胞也未产生影响。

基于CiteSpace知识图谱的何首乌致肝损伤研究热点与趋势分析

目的基于CiteSpace可视化工具对何首乌致肝损伤中英文文献进行可视化分析,了解和探索该研究领域的研究现状、热点和发展趋势。

何首乌提取物基于IL-23/IL-17轴对银屑病小鼠皮损及抑炎机制研究

目的 探讨何首乌提取物(PME)对银屑病小鼠皮损的改善及抑炎作用,并阐明其可能的作用机制。方法 雌性BALB/c小鼠随机分为正常组、模型组和PME低、中、高剂量组,每组各15只,除正常组外,剩余各组利用咪喹莫特(IMQ)诱导银屑病模型后给药,连续给药7 d。记录各组小鼠背部皮损情况并进行银屑病皮损面积及严重程度指数(PASI)评分;小鼠背部表皮组织行HE染色;免疫组化法检测CD4及K17的阳性表达;酶联免疫吸附测定(ELISA)法检测皮肤组织内肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)表达;实时荧光定量聚合酶链反应(RT-qPCR)检测IL-17A、IL-17F、IL-22、IL-23信使RNA(mRNA)相对表达水平;蛋白质印迹(Western blotting)法检测细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)蛋白相对表达量。结果 与正常组比较,模型组小鼠PASI评分升高(P<0.05);棘层增厚呈嵴状延长,有明显的细胞炎性水肿,皮肤表皮层厚度增大(P<0.05);CD4、K17阳性表达、TNF-α及IL-8含量、IL-17A、IL-17F、IL-22、IL-23 mRNA相对表达水平及ICAM-1、VCAM-1蛋白相对表达量均提高(P<0.05);与模型组比较,PME低、中、高剂量组小鼠银屑病PASI评分降低,炎性细胞浸润减少,棘层畸变改善,表皮层厚度降低(P<0.05);CD4、K17阳性表达、TNF-α及IL-8含量、IL-17A、IL-17F、IL-22和IL-23 mRNA相对表达水平及ICAM-1和VCAM-1蛋白相对表达量降低(P<0.05);PME作用效果呈剂量依赖性。结论 PME可改善IMQ诱导的银屑病小鼠皮损情况,可能与抑制IL-23/IL-17炎性轴有关。

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。