Using Network Pharmacology to Explore Therapeutic Effect of Polygonum capitatum on Renal Calculus in Rats

[Objectives]To explore the therapeutic effect of Polygonum capitatum on renal calculus in rats based on network pharmacology.[Methods]Through the preliminary construction of P.capitatum-urolithiasis disease target network,explore the active components,action pathway and action target of P.capitatum-urolithiasis treatment,and use 1%ethylene glycol+2%ammonium chloride to induce SD rat kidney calcium oxalate stone model to verify the efficacy of P.capitatum-urolithiasis treatment.[Results]Through the network pharmacological prediction,it is found that the important active components in P.capitatum were quercetin,gallic acid,rutin,silybin,catechin,kaempferol and so on;potential active targets include INS,CAT,IL-6,MOCOS,etc.The results also suggest that forkhead transcription factor signaling pathway(FoxO signaling pathway),tumor necrosis factor signaling pathway(TNF signaling pathway)and hypoxia-inducible factor signaling pathway(HIF-1 signaling pathway)are the core pathways.The results of biochemical indicators in animal experiment showed that the contents of serum urea nitrogen(BUN),creatinine(Cr)and malondialdehyde(MDA)in renal tissue in the treatment group(200,500 mg/kg)were significantly lower than those in the model group,while the content of superoxide dismutase(SOD)was significantly higher than that in the model group.In addition,the kidney tissue H&E staining sections showed that P.capitatum alcohol extract administration group rats kidney calcium oxalate crystals were significantly reduced compared with the model group,the degree of renal tubular lumen expansion was lighter than the model group,suggesting that P.capitatum alcohol extract has the effect of improving renal calculus in rats.[Conclusions]This study provides a theoretical reference for the deep development of P.capitatum in the treatment of renal calculus.

A New Flavonoid Glycoside from Polygonum capitatum

[Objectives]To discover novel compounds with significant anti-inflammatory activity in the alcohol extract of Polygonum capitatum.[Methods]Firstly,a new flavonoid glycoside,capitaone B(1),was isolated from the ethyl acetate extract from P.capitatum,and then 27 compounds were screened using NO anti-inflammatory activity model.[Results]Four compounds,such as kaempferol(12),1,2,6-trigalloyl-β-d-glucose(15),catechin(16)andβ-sitosterol(26),could significantly inhibit LPS-induced NO production in macrophages,with IC 50 values of 15.31,8.43,6.92 and 5.72μM,respectively.[Conclusions]The chemical composition and anti-inflammatory activity of P.capitatum were preliminarily studied,and the results provide a theoretical basis for further research on the action mechanism of subsequent anti-inflammatory active compounds of P.capitatum.

Screening for Antioxidant Phenolic Compounds from Polygonum capitatum

[Objectives]To discover antioxidant natural products from the famous Hmong medicinal plant Polygonum capitatum.[Methods]The antioxidant activities of the isolated components were evaluated by ABTS and DPPH assays.[Results]A total of 27 free phenolics were isolated form P.capitatum.Then the in vitro antioxidant potential of these components was evaluated according to the DPPH and ABTS radical scavenging assays.Among them,five compounds(13,14,17,23,and 25)showed most significant ABTS radical-scavenging activity(IC 50 values of 3.81-15.09μg/mL).And 12 components(1,2,6,7,9,12,13,14,16,17,23,and 25)showed notable radical scavenging activity against DPPH(inhibition rates>88%).[Conclusions]Most of the above bioactive compounds were reported for the first time.

Systematic Evaluation of Pharmacognostic Identification of Polygonum capitatum

[Objectives] To investigate the systematic evaluation of pharmacognostic identification of Polygonum capitatum . [Methods] 10 batches of P. capitatum cultivated in Guizhou were chosen for plant samples. Macroscopical identification was conducted on plant roots, stems, leaves, flowers and fruits. The P. capitatum powder was processed for physical and chemical distinction by FeCl 3 chromogenic reaction, hydrochloric acid magnesium powder reaction, AlCl 3 color development reaction and thin-layer chromatography.Microscope identification was carried out on the powder. Plant genome DNeasy Plant Kit was adopted for DNA molecular marker identification. [Results] The results showed that the stem of P. capitatum was tufted, the leaves were oval, 2 to 5 cm long, and 1 to 2 cm wide;the leaf apex was acute and cuneate at the base, the inflorescence was capitate, paired or solitary;the raceme was erect and nearly spherical, and the perianth was light red. Furthermore, for the chromogenic reaction of FeCl 3 ethanol extract of P. capitatum , appeared blue and turned to dark blue after long time storing at room temperature. For the reaction of hydrochloric acid magnesium powder, the alcohol extract of P. capitatum , exhibited deep red. In the color reaction of AlCl 3, the alcohol extract revealed yellow fluorescence under 360 nm UV lamp. Microscope identification of the powder displayed pollen grains, crystal sheath fibers, cellulose, vessels, starch grains, cork cells, and other characteristic fragments. In addition, DNA barcoding electrophoresis results showed that P. capitatum showed a clear and bright single band near 500 bp, and further sequencing results showed that the sequence differences were mainly concentrated in ITS1 and ITS2 region. [Conclusions] Systematic evaluation for the identification of P. capitatum is established, which combines with macroscopic identification, physicochemical identification, powder microscope identification, and DNA molecular identification. Finally, the original medicinal material is identified as P. capitatum Buch.-Ham. ex D. Don.

Antioxidant, anti-quorum sensing and anti-biofilm potential of ethanolic leaf extract of Phrynium capitatum and Dryptes indica

Objective: To investigate the antioxidant and anti-infective potential of Phrynium capitatum and Dryptes indica extract. Methods: The antioxidant potentials were determined by DPPH radical scavenging, reducing power, hydroxyl radical scavenging and total antioxidant assays. We further examined anti-quorum sensing activity and inhibition of synthesis of pathogenic factor of Chromobacterium violaceum and Pseudomonas aeruginosa PAO1. Bioactive compounds were determined using gas chromatography–mass spectrometry analysis. In silico analysis was conducted to determine the binding affinity of bioactive compounds of plant extracts for the quorum sensing regulatory receptor LasR. Results: DPPH assay showed that the ethanolic extract of Phrynium capitatum and Dryptes indica at 500 μg/mL showed (86.96 ± 4.07)% and (74.83 ± 3.47)% inhibition, respectively. Hydroxyl radical scavenging assay showed (73.17 ± 3.03)% and (62.63 ± 4.59)% activity, respectively. The ethanolic extract of Phrynium capitatum and Dryptes indica showed high level of attenuation of quorum sensing regulated pyocyanin production. Confocal laser scanning microscopic analysis revealed that the extracts had the potential to effectively inhibit biofilm formation of Pseudomonas aeruginosa. Molecular docking analysis showed a better binding affinity of bioactive compounds from the extracts for the structure of LasR protein of Pseudomonas aeruginosa. Conclusions: The ethanolic extracts of Phrynium capitatum and Dryptes indica possess antioxidant activity and the potential to inhibit the quorum sensing system and its regulatory virulence traits in Pseudomonas aeruginosa PAO1.

Study on the Pharmacological Identification of Polygonum capitatum

[Objectives]This study was conducted to establish a microscopic identification method of Polygonum capitatum Buch.Ham.ex D.Don.[Methods]The cross sections were identified by microscopic identification.[Results]The stem cross section of P.capitatum is round-like,and shows pericyclic fibers forming a ring,strongly lignified,many vascular bundles,and a hollow pith part.There are many starch granules in the powder,and single granules are more common;and fibers are mostly bundled or scattered,lignified or non-lignified,and calcium oxalate cluster crystals are common.There are many pollen grains,obtuse triangular or round-like,and some of them have three germination apertures.They have fine thorn-like protrusions on the outer wall,the surface of which has reticulate carvings.[Conclusions]The results of microscopic identification are reliable and can be used as the basis for identification of P.capitatum.

Antioxidant,Anti-inflammatory,and Antibacterial Activities of Different Extracts from Polygonum capitatum

[Objectives]To investigate the antioxidant,anti-inflammatory and antibacterial activities of different extracts(aqueous,ethanol,ethyl acetate and n-butanol extracts)of Miao medicine Polygonum capitatum.[Methods]Eleven batches of P.capitatum in Guizhou province were collected,and water,ethanol,ethyl acetate and n-butanol extracts were prepared by reflux extraction.Antioxidant activity was determined by radical scavenging capacity of 1,1 diphenyl-2-picyl hydrazine(DPPH),anti-inflammatory activity was screened by lipopolysaccharide(LPS)induced RAW264.7 cells to produce NO,and the minimum inhibitory concentration(MIC)was screened by broth microdilution method.[Results]When the concentration of ethyl acetate extract was 10 mg/L,the scavenging rate of DPPH ranged from 90%to 99%.The(MIC of the ethyl acetate extract against Staphylococcus aureus(SA),Pseudomonas aeruginosa(PA)and Escherichia coli(EC)was 0.18-0.65,0.13-0.82,and 0.15-0.78 g/L,respectively.In the anti-inflammatory activity,ethyl acetate extract inhibited NO production with inhibition rate of 70%.[Conclusions]The ethyl acetate extract and ethanol extract of Miao medicine P.capitatum have strong antioxidant,anti-inflammatory and antibacterial activities.

A Research Summary on the Chemical Constituents and Pharmacological Effects of the National Medicine Polygonum capitatum

Polygonum capitatum Buch.Ham.ex D.Don is a commonly used medicine in Zhuang people’s settlements,which has antipyretic,wind and evil-dispelling,dehumidifying and analgesic effects.The research on the chemical composition of P.capitatum began in the 1980 s.Modern scientific research has shown that it has antibacterial,cooling and blood sugar lowering effect.In order to better develop and utilize P.capitatum,this paper reviewed the progress of the chemical composition and pharmacological research of P.capitatum.

Identification of Ethnic Medicine Polygonum capitatum by Thin Layer Chromatography

[Objectives]To establish a thin-layer chromatography identification method for the ethnic medicinal materials of Polygonum capitatum.[Methods]The thin layer chromatography(TLC)method was used to identify materials.[Results]The TLC identification showed that in the chromatogram of the test sample,there were spots of the same color in the corresponding position of the chromatogram of the reference medicinal material,and the reproducibility was good.[Conclusions]The TLC identification and the determination results of the inspection items can provide a basis for the quality control of the medicinal materials of P.capitatum.

基于LC-MS和GC-MS代谢组学分析头花蓼不同入药部位黄酮类成分

为了探究头花蓼不同入药部位黄酮类化合物分布的丰度,挖掘其潜在的药用价值。采用LC-MS和GC-MS代谢组学技术对头花蓼茎、花和叶进行黄酮类化合物代谢组学比较研究。结果表明:头花蓼花中黄酮类化合物分布的数量(425个)>叶(376个)>茎(354个);以差异倍数(fold change)≥2或≤0.5,P<0.05为依据在头花蓼茎与花、茎与叶及叶与花对比中分别筛选到333个、289个和366个黄酮类差异代谢物。头花蓼三组样本对比(茎与花、茎与叶、叶与花)的差异代谢物显著富集于黄酮生物合成途径。3′,4′,5′,5,7-五羟黄酮、表儿茶素和表没食子儿茶素在花中的相对含量显著高于茎或叶,并且是重要的药理活性物质。本研究全面分析和鉴定了头花蓼不同入药部位的黄酮类化合物,证实了黄酮类化合物主要分布在花,为后续深入挖掘和利用其黄酮类化合物的药用价值提供理论基础。

种植年限对头花蓼土壤微生物群落的影响

【目的】探明不同种植年限头花蓼土壤微生物(细菌和真菌)群落组成及物种多样性的变化趋势,从土壤微生物的角度探究其连作障碍机制,为贵州头花蓼产业的可持续发展提供理论依据。【方法】采用Illumina测序技术,以头花蓼与玉米轮作地块土壤为对照(CK),通过对头花蓼连续种植2年(T2)、3年(T3)和4年(T4)地块土壤微生物进行测序分析,研究种植年限对头花蓼土壤微生物群落的影响。【结果】随连作年限增加,细菌群落丰度呈上升趋势,丰度从1620.30升至1787.22,物种多样性逐渐趋于平稳;真菌群落丰度呈下降趋势,丰度从635.00降至612.89,物种多样性变化不大。细菌群落:门水平下,变形菌门与酸杆菌门为优势菌群,相对丰度总占比>64%;变形菌门相对丰度显著降低(从40.80%降至34.80%),酸杆菌门显著增加(从24.20%升至29.10%);变形菌门(40.80%、40.20%、37.60%和34.80%)、浮霉菌门(1.50%、1.50%、2.60%和2.50%)在不同连作年限间相对丰度存在显著差异;属水平下,uncultured_bacterium_c_Subgroup_6所占比例最大,为10.66%~20.50%。真菌群落:门水平下,子囊菌门和担子菌门为优势菌群,其相对丰度总占比>61%,较CK逐年下降;被孢霉门和壶菌门在连作第4年(T_(4))几乎完全消失。属水平下,被孢霉属占比最大,为2.66%~14.30%,在连作第3年(T3)出现多孢囊霉属真菌。【结论】连作加大了头花蓼土壤中微生物群落差异。随着连续种植年限增加,变形菌门细菌、被孢霉门被孢霉属真菌等相对丰度逐渐降低,酸杆菌门细菌、子囊菌门镰刀菌属真菌等相对丰度逐渐增加,可能是头花蓼连作障碍发生的原因之一。

PB-BBD响应面法优化头花蓼提取工艺及抗氧化活性研究

目的利用Plackett-Burman和Box-Behnken响应面法优化头花蓼的总黄酮和总多酚超声提取工艺并研究其抗氧化活性。方法以头花蓼总黄酮和总多酚的综合得率为指标,通过单因素实验确定各影响因素的范围,再利用Plackett-Burman实验筛选影响综合得率的显著因素,最后采用Box-Behnken响应面法进行工艺优化;并通过考察提取物对DPPH自由基、羟基自由基的清除率和对Fe3+的还原能力来评价头花蓼的抗氧化性活性。结果影响综合得率的显著因素为超声功率、超声温度和超声时间,最优提取工艺为超声功率160W、超声温度58℃、超声时间38min、乙醇浓度40%、液料比25∶1(mL∶g),头花蓼中总黄酮和总多酚的综合得率为68.79mg·g^(-1),与模型预测值的相对误差为4.15%;当浓度为5mg·mL^(-1)时,此工艺条件下头花蓼提取物对DPPH自由基和羟基自由基的清除率分别为81.66%、84.93%,总还原能力达到2.48。结论采用基于PB实验设计和BBD响应面法优化得到的头花蓼总黄酮和总多酚提取工艺简便、稳定、可行,且得到的头花蓼提取物具有较强的抗氧化活性。

基于网络药理学与分子对接探索头花蓼抗肺炎的作用机制

目的:运用网络药理学与分子对接探索头花蓼抗肺炎的作用机制。方法:通过查阅文献收集头花蓼的化学成分,在chemdraw软件绘制分子结构图,通过PharmMapper数据库选择(NF)≥0.9的靶点蛋白获取潜在化合物的靶点;运用GeneCards、OMIM、DRUGBANK数据库以获得肺炎核心靶点;使用Metascape平台将头花蓼活性成分靶点和肺炎核心靶点交集进行GO和KEGG富集分析;使用Cytoscape3.8.2软件构建成分-靶点-通路图,AutoDock Tools 1.5.7、Pymol软件进行分子对接。结果:头花蓼中筛选获得38个活性成分、116个成分靶点、6个核心靶点;KEGG富集分析结果主要集中在IL-17信号通路、MAPK信号通路、TNF信号通路等;分子对接结果显示ALB、EGFR、CASP3、MAPK1、MAPK14、SCR与头花蓼主要化学成分的结合活性较好。结论:头花蓼中的核心化合物槲皮素、木犀草素、儿茶素等,可能通过ALB、EGFR、CASP3、MAPK1、MAPK14、SCR等靶点作用于IL-17、MAPK、TNF信号通路,从而达到治疗肺炎的作用。

头花蓼对多重耐药金黄色葡萄球菌抗菌作用研究

目的研究头花蓼对多重耐药金黄色葡萄球菌的抗菌活性。方法采用纸片扩散法测定抑菌圈直径,微量肉汤稀释法测定各实验菌株的最低抑菌质量浓度（MIC）和半数抑制质量浓度（IC50值）,液体转染法测定最低杀菌质量浓度（MBC）。结果头花蓼提取物对各多重耐药金黄色葡萄球菌均有不同程度的抑制作用,60%乙醇提取物对各耐药菌的抑菌圈明显大于水提物,并大于没食子酸。60%乙醇提取物与水提物对各耐药菌株的MIC值范围为0.78~3.12 mg/mL和IC50值范围为0.46~1.81 mg/mL,均小于没食子酸的MIC值和IC50值。60%乙醇提取物MBC值范围为1.56~6.25 mg/mL,小于水提物和没食子酸。结论头花蓼乙醇提取物体外抗多重耐药金黄色葡萄球菌作用优于没食子酸,提示头花蓼除了没食子酸外还存在其他抗菌成分。

头花蓼化学成分的研究(Ⅰ)

目的:研究头花蓼的化学成分。方法:采用反复硅胶柱层析和葡聚糖凝胶柱层析的色谱方法分离纯化头花蓼的化学成分,并根据理化性质和波谱数据进行结构鉴定。结果:从头花蓼的石油醚萃取部位分离鉴定了14个化合物,分别为二十三烷醇(Ⅰ),二十五烷醇(Ⅱ),二十二烷酸(Ⅲ),二十四烷酸(Ⅳ),十六烷酸-2,3-二羟基丙酯(Ⅴ),二十二烷酸-2,3-二羟基丙酯(Ⅵ),齐墩果酸(Ⅶ),乌苏酸(Ⅷ),二十八烷基-1,27-二烯(Ⅸ),阿魏酸二十二酯(Ⅹ),β-谷甾醇(Ⅺ),二十三烷(Ⅻ),十六烷酸(X),亚油酸(X)。结论:化合物Ⅰ～Ⅹ为从头花蓼中首次分离得到,化合物Ⅸ、Ⅹ为从蓼属中首次分离得到。

头花蓼对幽门螺杆菌粘附定植的影响

目的观察头花蓼水提物对幽门螺杆菌(Helicobacter pylori,H.pylori)粘附定植的影响。方法采用尿素酶活性比色法定量检测头花蓼水提物作用前后幽门螺杆菌尿素酶活性的影响;运用半固体布氏琼脂平板法检测不同浓度头花蓼对幽门螺杆菌鞭毛运动的影响;利用细胞爬片革兰染色观察不同浓度头花蓼作用后幽门螺杆菌在GES-1细胞表面粘附的变化并计算各组抑制率;采用Real-time PCR技术检测头花蓼作用前后相关基因ureB、ureE、ureF、flaA、flaB、babA、alpA、alpB的mRNA转录水平。结果头花蓼水提物对幽门螺杆菌尿素酶活性具有抑制作用,抑制率为44.90±0.289。头花蓼浓度为1/2MIC、1/4MIC、1/8MIC分别作用后幽门螺杆菌菌膜晕圈直径显著小于对照组,分别为(5.67±0.55)mm,(8.5±0.50)mm,(11.67±1.53)mm,差异有统计学意义(P<0.05);不同浓度头花蓼水提物均能抑制细菌粘附胃上皮细胞,各浓度组与对照组比较,差异有统计学意义(P<0.05)。头花蓼作用后,与对照组比较,下调了尿素酶结构基因ureB,尿素酶活性基因ureE、ureF、鞭毛相关基因flaB、flaA以及粘附素基因babA、alpA、alpB mRNA的转录水平,差异均具有统计学意义(P<0.05)。结论头花蓼对幽门螺杆菌具有明显的抑制作用,是通过抑制幽门螺杆菌尿素酶活性,减缓幽门螺杆菌的鞭毛动力,影响幽门螺杆菌对胃粘膜上皮细胞的粘附等方面发挥其抑菌作用。

头花蓼化学成分及抗氧化活性研究

目的：对头花蓼的化学成分及其生物活性进行研究,为阐明有效成分提供依据。方法：利用硅胶、聚酰胺和Sephadex LH-20对其化学成分进行分离纯化,根据化合物的理化性质及光谱数据鉴定其结构。采用化学发光法测试化合物的抗氧化活性。结果：从该植物全草中分得10个化合物,分别鉴定为没食子酸（1）,原儿茶酸（2）,没食子酸乙酯（3）,山柰酚（4）,槲皮素（5）,槲皮苷（6）,阿魏酸二十四烷酯（7）,槲皮素-3-O-β-D-葡萄糖苷（8）,β-胡萝卜苷（9）和β-谷甾醇（10）。在药理活性研究中,化合物1～6在不同的药理模型中均显示了一定的抗氧化活性。结论：化合物3和7是从该属植物中首次分离得到,化合物2、4和9为该植物中首次分离得到。化合物1-6均具有一定的抗氧化作用,这也是首次对该植物的单体成分进行药理活性研究。

HPLC/DAD/MS法同时测定苗药头花蓼中3种有效成分的含量

目的:建立HPLC法测定头花蓼中没食子酸、davidiin和槲皮苷的含量。方法:采用 Alltima C_(18)色谱柱(150 mnm×4.6mm,5μm),以A:2.5%醋酸水溶液,B:水-四氢呋喃-甲醇(40:10:50)为流动相进行梯度洗脱,梯度条件如下:0 min,80:20;15 min,10:90;20 min,10:90;25 min,80:20;30 min,80:20。流速为0.6 mL·min^(-1),紫外检测波长为270 nm。离子源APCI,扫描模式负离子。结果:没食子酸、davidiin和槲皮苷的线性范围(n=6)分别为0.017～2.28 μg(r=0.9999),0.05～2.50μg(r=0.9999),0.16～5.10 μg(r=0.9999);平均回收率(n=9)分别为98.0%,99.6%,98.5%。提取方法为80%乙醇冷浸12 h后超声30 min。结论:本方法简便、准确,可为评价不同产地的头花蓼质量提供依据。

HPLC测定不同产地的头花蓼中没食子酸的含量

目的测定不同产地头花蓼中没食子酸的含量。方法采用依利特Hypersil C18色谱柱(150 mm×4.6 mm,5μm),以乙腈-0.4%磷酸(5:95)为流动相,流速0.8 ml.min-1,检测波长272 nm。结果没食子酸的线性范围为0.0816-0.4896μg(r=0.9996),平均回收率为97.7%,RSD=0.6%。结论所建方法准确、简便,可评价不同产地头花蓼质量。

头花蓼对重金属Cd的吸收特性与累积规律初探

以野生地被植物头花蓼为试验材料,采用温室盆栽法,对重金属Cd在该植物体内的吸收、累积分布以及迁移特性进行了初步研究。结果表明,Cd对头花蓼生长未造成显著影响,甚至低浓度(≤5mg.kg-1)Cd具有一定的生长促进作用,表现为植物的生物量增加。当Cd处理浓度达到50mg.kg-1时,植物的生物量虽有所降低,但与对照相比并无显著差异。植株不同部位对Cd的积累具有分异特性,地下部根系的累积量最大,叶次之,茎最小,且随处理浓度的增加而增加,在Cd处理浓度为50mg.kg-1时均达到最大值,分别为182.69、31.49mg.kg-1和10.34mg.kg-1。植株对Cd富集系数和转移系数分别为0.46～1.55和0.14～0.67,且地上部对Cd的最大迁移总量高达100.09μg.plant-1。说明头花蓼对修复Cd污染土壤具有一定的潜力,是一种修复Cd污染较好的景观地被植物种质资源。