Exploring the mechanism of action of herb pair Pinellia Ternata-Magnolia Officinalis in the treatment of liver cancer based on network pharmacology and molecular docking

Background:Explore the anti-tumor mechanism of herb pair Pinellia ternate-Magnolia officinalis(BX-HP)in liver cancer through network pharmacology using molecular docking methods.Method:The active ingredients and corresponding targets of the herb pair Pinellia ternate-Magnolia officinalis were obtained from the HERB database.The relevant targets for liver cancer were obtained from GeneCards,DisGeNET,TTD,and Drugbank databases.Obtain common targets between herb pair Pinellia ternate-Magnolia officinalis and liver cancer through the Bioinformatics platform,establish a PPI network diagram using STRING software,and perform GO functional enrichment and KEGG pathway enrichment analysis on the DAVID platform.AutoDockTools 1.5.7 software and molecular dynamics simulation analysis are used to evaluate the binding of components to target proteins.HERB database,SwissTargetPrediction database,SwissADME database,UniProt database,GeneCards database,TTD database,DRUGBANK database,DisGeNET database,String,DAVID.Bioinformatics platform,PDB database,PubChem and TCMSP database.Result:A total of 22 active ingredients with a Probability>0.1 targets in Magnolia officinalis were screened,26 active ingredients with a Probability>0.1 targets in Pinellia ternata,ten vital active ingredients,corresponding to 979 and 803 targets with a Probability>0.1 targets,2536 liver cancer-related targets,and 279 targets in the herb pair Pinellia ternata-Magnolia officinalis.The GO functional enrichment analysis resulted in 1297 entries,namely 971 biological process entries,118 cell localization entries,and 208 molecular function entries.Three signaling pathways were annotated through the KEGG pathway.Based on molecular docking,ten vital active ingredients and five target proteins were validated to exhibit an excellent binding affinity.The above data indicates that combining the herb pair Pinellia ternata-Magnolia officinalis may treat liver cancer through specific targets and signaling pathways.Conclusion:Herb pair Pinellia ternata-Magnolia officinalis has a synergistic effect on treating liver cancer through multicomponent,multitarget,and multi-pathway approaches.This study provides a sufficient theoretical basis for subsequent research.

Oxalic acid in ginger specifically denatures the acrid raphides in the unprocessed dried tuber of Pinellia ternata

Objective:Pinellia Tuber,the dried tuber of Pinellia ternata,is widely used in Japanese Kampo medicines and traditional Chinese medicines.The unprocessed Pinellia Tuber is known to cause very strong acrid irritation at oral and laryngopharynx mucosa.Recent studies have shown that the sharp needle-like crystals called raphides,that are composed of calcium oxalate and proteins,are the main causative substances of the irritation.Ginger,the rhizome of Zingiber officinale,has been used in the processing to reduce the acridity of Pinellia Tuber since before the sixth century,however,the mechanisms of reducing acridity have not been scientifically proved yet.Methods:We developed the raphides denaturation assay(RDA)to quantify the degree of denaturation in the raphides to cause irritation.By their lipophilic characters,the raphides could be extracted in petroleum ether(PE)layer from powdered Pinellia Tuber suspended in water,and the contents of the raphides in PE layer were measured by the absorbance.By this assay,we conducted the activity-guided fractionation from the boiling water extract of ginger to find the ingredients to denature the raphides.We also conducted the gustatory tests to detect the change of the irritation of the denatured raphides.Results:The treatment of powdered Pinellia Tuber suspension with ginger extract reduced the distribution of raphides in PE layer in RDA in a concentration-dependent manner.The activity-guided fractionation using RDA revealed that oxalic acid was the main active ingredient in ginger extract to denature the raphides of Pinellia Tuber.Oxalic acid reduced the lipophilicity of the raphides in the thermo-,time-,and concentration-dependent manners,and its activity was affected by pH.The treatment of powdered Pinellia Tuber suspension with oxalic acid significantly reduced its acrid irritation in gustatory test in human.Conclusions:We found that oxalic acid is the main active ingredient in ginger to reduce the acrid irritation of Pinellia Tuber.

三叶半夏(Pinellia ternata)雄配子败育的遗传分析

从减数分裂过程，小池子发育和染色体数目与组成三方面探讨了三叶半夏雄配子败育的原 因。三叶半夏（Ｐｉｎｅｌｌｉａ ｔｅｒｎａｔａ）体细胞染色体数目２ｎ＝９０，其减数分裂异常，多数同源染色体能配对形 成二价体，部分同源染色体呈单价体形式或配对成多价体。减数分裂的各时期都能观察到导致遗传物 质不均衡分离的异常结构，如落后染色体，染色体桥和微核等。根据结果推测：三叶半夏是具有染色体 结构变异的多倍体复合种。染色体结构变异是三叶半夏雄配子败育的主要原因。小孢子母细胞都能完 成减数分裂过程，且单核小孢子能通过两次有丝分裂形成三核花粉粒。成熟花粉粒中可育花粉不足 １％。三核期是雄配子败育的时期。

半夏属(PINELLIA)植物的叶绿素含量和光合速率日变化

对半夏属５个种７个群体植株的叶片进行了叶绿素含量测定和净光合速率日变化比较，发现半夏Ｐｔｅｒｎａｔａ（Ｔｈｒｎｂ）Ｂｒｅｉｔ和虎掌Ｐ．ｐｅｄａｔｉｓｅｃｔａＳｃｈｏｔ都较耐强光，但半夏的净光合速率较高。倒苗是半夏避旱的适应方式，鹞落坪半夏Ｐ．ｙａｏｌｕｏｐｉｎｇｅｎｓｉｓＸ．Ｈ．Ｇｕｏｅｔ．Ｘ．Ｌ．Ｌｉｕ适应性差，应就地保护。

Quality evaluation of Pinelliae Rhizoma using network pharmacology and multi-component quantitative analysis

Background:Pinelliae Rhizoma(Banxia)is a valuable traditional Chinese medicine,and its quality issues are related to the safety and effectiveness of the medicine.Several pharmacological experiments have shown that Pinelliae Rhizoma has anti-inflammatory activity,but the specific chemical components remain unclear.Methods:In the present study,network pharmacology was used to analyze the potential active ingredients and molecular mechanisms of Pinelliae Rhizoma’s anti-inflammatory activity.A new approach to simultaneously determine eight components using high performance liquid chromatography-photo-diode array was developed to evaluate the quality of Pinelliae Rhizoma and different processed Pinelliae Rhizoma products.Results:Twelve active ingredients were identified from Pinelliae Rhizoma,andβ-sitosterol may have a greater effect than the other active ingredients.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed the main pathways associated with Pinelliae Rhizoma’s mechanism for treating inflammation.Additionally,theβ-sitosterol content in different processed Pinelliae Rhizoma products decreased significantly,and the level of five nucleosides in Pinelliae Rhizoma was significantly higher than that of the processed Pinelliae Rhizoma products.The 6-gingerol content was detected in eight Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine batches with different origins,and liquiritin and liquiritigenin levels were detected in eight batches of Pinelliae Rhizoma Praeparatum with different origins.Conclusion:Pinelliae Rhizoma quality was affected by different regions and different processing methods,and this research provides a reference for Pinelliae Rhizoma quality evaluation.

Application of Metabolomics Approach to Study on Chemical Constituents in Cultivated and Wild Pinellia ternate (Thunb.) Breit. and Typhonium flagelliforme (Lodd.) Blume Based on UPLC-ESI-HRMS^n

[Objectives] The aim was to study chemical components of cultivated and wild Pinellia ternate(Thunb.) Breit and Typhonium flagelliforme(Lodd.) Blume by metabolomics technology based on UPLC-ESI-HRMS^n. [Methods] Chemical components of 27 samples of cultivated and wild P. ternate and T. flagelliforme were detected by using UPLC-ESI-HRMS^ntechnology. Differential markers between P. ternate and T. flagelliforme as well as cultivated and wild P. ternate were screened and identified by principal component analysis(PCA) and orthogonal partial least square-discriminate analysis(OPLS-DA). [Results]12 components of P. ternate(cultivated and wild variety) and T.flagelliforme and three components of cultivated and wild P. ternate were screened and identified by OPLS-DA. N-Acetyl-6-hydroxy-L-tryptophan and N-Phenylacetylglutamic were not detected in P. ternate,and these two components can be used as potential markers of P. ternate and T. flagelliforme. [Conclusions]UPLC-ESI-HRMS^n-based metabolomics technology could be used to quickly identify the chemical constituents of P. ternate and effectively distinguish P. ternate and T. flagelliforme,which has reference significance for the quality control of P. ternate.

Research Progress of the Wild Medicinal Plant,Pinellia ternata

Based on literature reviews and analysis of research reports on Pinellia ternata found locally and abroad in recent years,this article summarizes and arranges them.The research on Pinellia ternata mainly focuses on its cultivation,tissue culture,and so on.There are only a few research on its active components and its regulation mechanism.The wild resources of Pinellia ternata are gradually decreasing,hence it is urgent to take effective measures to protect these wild resources as well as to establish germplasm resources bank and nursery.In order to meet the needs of the domestic market,it is necessary to investigate the distribution of wild Pinellia ternata resources,explore the best growing environment and conditions,artificially cultivate Pinellia ternata,as well as implement resource industrialization,sustainable development,and utilization.

Coptis,Pinellia,and Scutellaria as a promising new drug combination for treatment of Helicobacter pylori infection

BACKGROUND Helicobacter pylori(H.pylori)is the most important infectious agent and plays an important role in the progression of chronic gastritis and the development of gastric cancer.AIM To identify efficient therapeutic agents or strategies that can treat H.pylori infection.METHODS We performed literature analysis,experimental validation,and network pharmacology.First,traditional Chinese medicine(TCM)prescriptions for the treatment of H.pylori infection were obtained from the China National Knowledge Infrastructure,China Biology Medicine,China Science and Technology Journal Database,and WanFang databases.In addition,we conducted a relevant search by Reference Citation Analysis(RCA)(https://www.referencecitationanalysis.com).Next,we used TCM Inheritance Support System V2.5 to identify core drug combinations in the TCM prescriptions.Then,an H.pylori-associated chronic mouse model of gastritis was established.The antibacterial properties and antiinflammatory potential of the core drug combination were evaluated by the rapid urease test,modified Warthin-Starry silver staining,histopathological analysis,and enzyme linked immunosorbent assay.Finally,the active compounds,hub targets,and potential signaling pathways associated with the core drug combination were analyzed by network pharmacology.RESULTS The TCM treatment of H.pylori was mainly based on reinforcing the healthy Qi and eliminating pathogenic factors by simultaneously applying pungent dispersing,bitter descending,cold and warm drugs.The combination of Coptis,Pinellia,and Scutellaria(CPS)was identified as the core drug combination from 207 prescriptions and 168 herbs.This drug combination eradicated H.pylori,alleviated the gastric pathology induced by H.pylori infection,and reduced the expression levels of tumor necrosis factor-α(P=0.024)and interleukin-1β(P=0.001).Moreover,a total of 35 compounds and 2807 targets of CPS were identified using online databases.Nine key compounds(tenaxin I,neobaicalein,norwogonin,skullcapflavone II,baicalein,5,8,2'-trihydroxy-7-methoxyflavone,acacetin,panicolin,and wogonin)and nine hub target proteins(EGFR,PTGS2,STAT3,MAPK3,MAPK8,HSP90AA1,MAPK1,MMP9,and MTOR)were further explored.Seventy-seven signaling pathways were correlated with H.pylori-induced inflammation and carcinogenesis.CONCLUSION In summary,we showed that CPS is the core drug combination for treating H.pylori infection.Animal experiments demonstrated that CPS has bacteriostatic properties and can reduce the release of inflammatory cytokines in the gastric mucosa.Network pharmacology predictions further revealed that CPS showed complex chemical compositions with multi-target and multipathway regulatory mechanisms.Although the results derived from network pharmacology are not necessarily comprehensive,they still expand our understanding of CPS for treating H.pylori infection.

Characterization and biological activities of lectin isolated from <i>Pinellia ternata</i>

In this paper, Pinellia ternata lectin (PTL) purified from 95% saturated ammonium sulfate precipitation by applying mannose-Sepharose 4B affinity column chromatography was first described. The mannose-Sepharose 4B affinity adsorption gel has already been designed and prepared before the experiments according the combinative characterization between lectin and mannose. Mass spectrometry analysis result shows that PTL is a glycoprotein with the molecular weight of 12.165 kD. It was conclude that PTL had strong agglutination effects on mouse red blood cells, and the minimum reaction concentration was 25 μg/ml according the Hemagglutination. Results of automatic amino acid analysis indicated that PTL mainly contained 15 varieties of amino acids, of which the minimum content was cysteine and aspartate was the maximum. Cell experiment results suggested that PTL of low concentrations (0.004 mg/ml, 0.02 mg/ml and 0.1 mg/ml) promoted HeLa cell proliferation, but the effect weakened with the concentrations and treated-time increasing. However, the HeLa cells pro- liferation was intensively inhibited by higher PTL concentrations (0.5 mg/ml and 1 mg/ml), and the effect increased in a dose and time dependent manner.

Mechanism of Pinelliae Rhizoma in Treatment of Insomnia Based on Network Pharmacology

[Objectives]To explore the mechanism of Pinelliae Rhizoma in the treatment of insomnia.[Methods]Using the Traditional Chinese Medicine Systems Pharmacology(TCMSP)database,the effective components of Pinelliae Rhizoma were explored,to predict its targets,and screen out the targets of insomnia through GeneCards,OMIM and other databases.With the aid of Cytoscape software,a"component-target-pathway-disease"network for the treatment of insomnia by Pinelliae Rhizoma was built.Then,a target protein-related interaction network was created through the String database,and biological function annotation and pathway analysis of key targets were performed.[Results]The 13 active components of Pinelliae Rhizoma take action in the treatment of insomnia by intervening 33 targets,59 GO biological processes,and 10 main biological pathways.[Conclusions]The active components of Pinelliae Rhizoma may treat insomnia by such pharmacological effect as regulating G protein-coupled amine receptor activity,adrenergic receptor activity,catecholamine binding,and neurotransmitter receptor activity.

Mechanism of "Pinelliae rhizoma-Arisaematis rhizoma" in treatment of lung cancer based on network pharmacology

Objective:The study was designed to explore the mechanism of“Pinelliae RhizomaArisaematis Rhizoma”(PR-AP)in treatment of lung cancer based on network pharmacology.Methods:The active components and their targets of“Pinelliae Rhizoma-Arisaematis Rhizoma”were screened out from TCMSP database.The cytoscape3.7.0 software was used to construct the drug-active component-targets network.Searched the DisGeNet database to obtain lung cancer related targets.Bisogenet was used to construct"PPI network of active component target"and"PPI network of lung cancer target",and the intersection of the two networks was taken and the target was screened.AutoDockTools software was used to dock the key active ingredients with the key targets.David database was used to perform Go biological process enrichment analysis and KEGG pathway enrichment analysis.Results:A total of 15 active components were collected,including beta-sitosterol,baicalein,Stigmasterol,Cavidine,coniferin,etc.These active components mainly act on 95 key targets such as ESR1 and CDK2,which enriched in multiple lung cancer related signaling pathways,such as PI3KAkt、MAPK、HIF-1、FoxO、TGF-β、Hippo、TNF、Notch、VEGF、cAMP,etc.The results of molecular docking showed that the active components could regulate the core targets(ESR1 and CDK2).Conclusion:The effect of“Pinelliae Rhizoma-Arisaematis Rhizoma”in the treatment of lung cancer embodies the characteristics of multi-component,multi-target and multi-pathway of traditional Chinese medicine.Its mechanism of action may be to play a role in the treatment of lung cancer by regulating cell cycle,angiogenesis,tumor stem cells,etc.This research can provide ideas and references for further research.

Cloning and molecular characterization of a novel lectin gene from Pinellia ternata

The full-length cDNA of Pinellia ternata agglutinin (PTA) was cloned from inflorescences using RACE-PCR. Through comparative analysis of PTA gene (pta) and its deduced amino acid sequence with those of other Araceae species, pta was found to encode a precursor lectin with signal peptide and to have extensive homology with those of other Araceae species. PTA was a heterotetrameric mannose-binding lectin with three mannose-binding boxes like lectins from other Araceae and Amaryllidaceae species. Southern blot analysis of the genomic DNA revealed that pta belonged to a low-copy gene family. Northern blot analysis demonstrated that pta constitutively expressed in various plant tissues including root, leaf, stem and inflorescence. The pta cDNA sequence encoding for mature PTA protein was cloned into pET-32a plasmid and the resulting plasmid, pET-32a-PTA containing Trx-PTA fusion protein, was investigated for the expression in E. coli BL21. SDS-PAGE gel analysis showed that the Trx-PTA fusion protein was successfully expressed in E. coli BL21 when induced by IPTG. Artificial diet assay revealed that PTA fusion protein had significant levels of resistance against peach potato aphids when incorporated into artificial diet at 0.1% (w/v). The cloning of the pta gene will enable us to further test its effect in depth on aphids by transferring the gene into crop plants.

Study of dual-directional regulatory effect of Banxia （Pinellia ternata） and Huanglian （Coptis chinensis） drug pair on gastrointestinal movement of mice

摘要目的：观察半夏-黄连药对对功能性消化不良小鼠胃肠运动的调节作用.方法：采用三种不同配比的半夏-黄连药液（1：1,3：1,4：1）予昆明种雄性小鼠灌胃.10天后用阿托品腹腔注射法复制胃肠运动抑制模型,新斯的明腹腔注射法复制胃肠运动亢进模型,并且检测其小肠推进率.此外,使用与第一个实验相同的建模方法,对半夏-黄连3：1配比组设立高、中、低三种剂量浓度（4.68g/L,2.34g/L和1.17g/L）.运用酶联免疫吸附试验检测小肠组织内胃肠激素如胃泌素、血管活性肠肽和生长抑素的含量.结果：在新斯的明诱导的胃肠运动亢进模型中,3种不同配比的半夏-黄连药液（1：1,3：1,4：1）均能显著降低小鼠的小肠推进率（P=0.003,P=0.012,P=0.021）;在阿托品诱导的胃肠运动抑制模型组中,仅3：1配比的半夏-黄连药液能显著增加模型小鼠的小肠推进率.在新斯的明诱导的胃肠运动亢进模型中,半夏-黄连药液低剂量组、中剂量组和高剂量组的胃泌素水平均低于模型组（P=0.001,P=0.004,P=0.003）.半夏-黄连药液中剂量组和高剂量组的血管活性肠肽水平均高于模型组（P=0.004,P=0.002）.半夏-黄连药液中剂量组的生长抑素水平高于模型组（P=0.002）;在阿托品诱导的胃肠运动抑制模型中,半夏-黄连药液中剂量组和高剂量组的胃泌素水平均高于模型组（P=0.007,P=0.021）.半夏-黄连药液低剂量组,中剂量组和高剂量组的血管活性肠肽水平均高于模型组（P=0.001,P=0.001,P=0.001）.半夏-黄连药液中剂量组的生长抑素水平低于模型组（P=0.001和P=0.006）.结论：半夏-黄连可以通过影响小肠组织内的胃泌素、血管活性肠肽和生长抑素的水平来双向调节由阿托品及新斯诱导的小鼠功能性消化不良.

Physiological and transcriptional responses to heat stress in a typical phenotype of Pinellia ternata

Pinellia ternata is an important medicinal plant,and its growth and development are easily threatened by high temperature.In this study,comprehensive research on physiological,cytological and transcriptional responses to different levels of heat stress were conducted on a typical phenotype of P.ternata.First,P.ternata exhibited tolerance to the increased temperature,which was supported by normal growing leaves,as well as decreased and sustained photosynthetic parameters.Severe stress aggravated the damages,and P.ternata displayed an obvious leaf senescence phenotype,with significantly increased SOD and POD activities(46%and 213%).In addition,mesophyll cells were seriously damaged,chloroplast thylakoid was fuzzy,grana lamellae and stroma lamellae were obviously broken,and grana thylakoids were stacked,resulting in a dramatically declined photosynthetic rate(74.6%).Moreover,a total of 16808 genes were significantly differential expressed during this process,most of which were involved in photosynthesis,transmembrane transporter activity and plastid metabolism.The number of differentially expressed transcription factors in MYB and bHLH families was the largest,indicating that these genes might participate in heat stress response in P.ternata.These findings provide insight into the response to high temperature and facilitate the standardized cultivation of P.ternata.

金水六君煎物质基准中半夏和陈皮的薄层鉴别研究

目的:建立经典名方金水六君煎物质基准中半夏和陈皮的薄层鉴别方法。方法:以半夏对照药材为对照,对金水六君煎中半夏进行薄层鉴别研究,以橙皮苷和陈皮对照药材为对照,对金水六君煎中陈皮进行薄层鉴别研究,并对其进行方法学验证。结果:建立的薄层色谱法能鉴别出半夏和陈皮,斑点对应较好,阴性无干扰,方法学考察结果显示,方法适应性良好。结论:本法简单可行、结果可靠、重复性好,可作为金水六君煎物质基准的质量控制方法。

半夏曲传统制作工艺沿革探讨

该文对半夏曲传统制作工艺的沿革进行了探讨。宋代半夏曲的制作方法包括半夏姜汁制饼与制饼后发酵两种,以半夏姜汁制饼为主流。金元时沿袭宋代工艺,半夏曲的应用具有一定的局限性。明代医家对半夏曲的炮制机理进行了阐发,认为“片则刀峻,曲则力柔。”明代李时珍对半夏饼与半夏曲的混淆问题进行了澄清,韩懋的《韩氏医通》收载了5首半夏曲方,对半夏曲配方的扩充具有划时代的意义。清代医家在继承《韩氏医通》半夏曲方的基础上,总结为韩飞霞制曲十法;当时的半夏曲与神曲的制作工艺相互影响。建国后,半夏曲的地方炮制经验深受清代影响,地方炮制规范逐渐趋同。该文同时对半夏曲炮制工艺中的“半夏”与“曲”孰重孰轻、半夏的前期处理、半夏饼与半夏曲的概念区分、面粉用量与添加时机、神曲粉的加入、白矾在半夏曲中的作用等问题进行了思考。

姜制半夏传统制作工艺考证

姜半夏传统制作工艺:完整半夏姜制法多在前期生半夏汤洗的基础上进行,亦有直接用姜汤洗的记载。半夏片为汤洗后切片,再用姜汁浸、炒、煮、蒸等,以炒法为优。半夏粉为半夏汤洗切片研细粉,再用姜汤浸澄,多用于制作糊丸或包衣。半夏饼为汤洗后半夏研粉与生姜汁或去皮生姜混合制成的饼状物,再经炙、煨、炒等法而成。半夏曲始于宋代,当时半夏曲有制饼与发酵两种,均以生姜为辅料。制饼者并没有发酵的过程,实为半夏饼。半夏曲的配方众多,生姜必不可少。地方炮制经验中将生半夏水浸泡,生姜煮取汁,再共同煮制为主流。地方炮制规范中多为生半夏水浸透,姜汤矾水共煮。现代学者的研究方向主要集中在炮制工艺优选与炮制原理两方面。建议恢复姜半夏的传统汤洗姜制工艺。

息半夏总生物碱的提取工艺优化及其生物活性分析

以息半夏的块茎为原料,乙醇为超声波辅助提取溶剂,考察乙醇体积分数、超声时间、料液比、超声温度等单因素对总生物碱提取率的影响,设计正交试验优化提取工艺,并测试总生物碱的体外抗氧化和降血糖活性。结果表明,最佳提取条件为乙醇体积分数65%、提取时间60 min、料液比1∶45、超声温度65℃,总生物碱的提取率达0.502%;总生物碱对DPPH、ABTS自由基的清除率分别达89.90%和43.60%,对α-葡萄糖苷酶的抑制率达86.40%。该工艺简单可行,总生物碱表现出良好的体外抗氧化和降血糖生物活性。

半夏提取物对淡色库蚊幼虫的杀虫活性

通过不同萃取相(石油醚、氯仿、乙酸乙酯、正丁醇和水)提取半夏(Pinellia ternata)的活性物质,采用浸液法测定半夏提取物对淡色库蚊(Culex pipiens pallens)幼虫的杀虫活性。结果表明,半夏甲醇提取物对淡色库蚊幼虫具有显著的毒杀活性;在0.5 mg·mL^(-1)浓度处理下,石油醚、氯仿萃取相对淡色库蚊幼虫具有显著的杀虫活性,致死率在50%以上。对石油醚、氯仿萃取相进行室内毒力测定,LC50分别为0.21和0.31 mg·mL^(-1)。研究结果可为新型植物源农药综合利用提供参考。