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.

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.

Effects of extracts from Chuanwu(Aconitum Carmichaelii) and Banxia(Rhizoma Pinelliae) on excisional wound healing in a rat's model

OBJEVTIVE: To investigate the effects of extracts from Chuanwu(Aconitum Carmichaelii) and Banxia(Rhizoma Pinelliae) on the excisional wound healing in a rat's model.METHODS: Rats were performed a surgical lesion with a 2.0 cm resected tissue in the dorsal fascia.Following, animals were randomly divided into model group, YNB group and APE group those were respectively treated with saline, 1 mg/mL of Yunnan Baiyao and 1 mg/mL of Chuanwu(Aconitum Carmichaelii) and Banxia(Rhizoma Pinelliae) extracts. Wound contractions in days 0, 3, 7, 14 and 21 were calculated by an image analyzer. Histological analysis was analyzed using hematoxilin and eosin.Levels of tumor necrosis factor-2(IL-2), IL-4, IL-10, transformαin(TNF-g growα), interleukinth factor-1(TGF-nd basic fibroblast growth factor(bFββ1) a-GF) were determined by real-time quantitative PCR.RESULTS: Compared with that of YNB group and APE gtoup, the skin of rats showed poor re-modeling and re-epithelization characterized by a significant decrease of neovascularization, epithelialization and fibroblast in the model group. In the APE group, levels of TNF-α-and IL-2 were significantly down-regulated and IL4 and IL-10 significantly upregulated in contrasted with that of model group.In addition, levels of TGF-roup were significantly iβ1 and bFGF in the APE gnduced compared with that of model group.CONCLUSION: The results suggest that the extracts from Chuanwu(Aconitum Carmichaelii) and Banxia(Rhizoma Pinelliae) promote wound healing in the rats, which is associated with enhancing antiinflammatory ability and inducing fibroblast formation.

施肥对半夏生长·产量和质量的影响研究

[目的]研究施肥对半夏生长状况、产量和质量的影响,为半夏的规范化栽培提供依据。[方法]试验主要针对过磷酸钙的施肥水平进行研究,按施肥量和用种量的配比设4个处理,收获后测定产量和主要成分含量,研究不同施肥水平对半夏产量和质量的影响。[结果]施肥水平对半夏的生长影响差异不明显,对半夏质量影响差异明显。肥料1和4的半夏产量、净增重和增重率之间的差异不大,但明显高于肥料2和3;不同施肥水平半夏药材的水分、灰分、水浸物含量和总酸含量差异均不显著(P>0.05),而肥料1和肥料3半夏的总生物碱含量高于其他2个处理。[结论]施用过磷酸钙对半夏产量和质量影响不明显,因此生产上不使用化肥,以农家肥作底肥就能满足半夏生长需要。

半夏中β-谷甾醇的含量测定

[目的]建立高效液相色谱法测定半夏中β-谷甾醇的含量。[方法]色谱条件:色谱柱为岛津Shim-Pack VP-ODS C18(4.6 mm×150 mm,5μm),流动相为无水甲醇,流速为1.0 ml/min,检测波长为210 nm,柱温为35℃。[结果]β-谷甾醇在0.200～2.000μg/ml浓度范围内线性关系良好,r=0.999 6,平均回收率为101.07%(n=5),RSD为2.96%;半夏样品中β-谷甾醇含量的平均含量为1.59mg/g。[结论]该方法分离度较好,可用于半夏药材中β-谷甾醇的含量测定。

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.

油樟精油对半夏开放式组培愈伤组织诱导及分化的作用

以半夏(Pinellia ternata)叶柄、叶片作为外植体,采用组培方法研究不同浓度油樟精油对半夏愈伤组织诱导及分化的作用。结果表明,在基本培养基MS配方基础上,TDZ 0.3 mg/L和油樟精油体积浓度为0.20 mL/L的配方组合,半夏叶柄及叶片愈伤组织诱导率分别为92.3%、95.7%;TDZ 0.3 mg/L和油樟精油体积浓度为0.10 mL/L的配方组合时,分化率为100.0%,平均丛生苗数达5.25个,分化效果最佳。发现油樟精油体积浓度为0.20 mL/L时的促进效果相对较好,可以有效促进愈伤组织的诱导,提高愈伤诱导率,愈伤组织质地较紧密,呈浅绿色,增殖效果较好,分化率达100%,愈伤组织出现的芽点相对明显,且数量相对较多。综合培养成本来看,油樟精油在半夏开放式组培体系的最适浓度为0.20 mL/L。

Synergistic effect of pinellia total alkaloids and uncaria total alkaloids on anticonvulsant action in mice and rats

Aim To investigate the synergistic effect of the combination of pinellia total alkaloid （PTA） and uncaria total alkaloid （UTA）, and explore the mechanism of anticonvulsant action. Methods Anticonvulsant and toxic effect profiles of combinations of PTA with UTA, alone and at three fixed ratios of 1：4, 1 ：1, 4：1, were evaluated in maximal electroshock （MES）-induced seizures and acute toxicity test in mice. Respective ED50 and LD50 were calculated with Bliss＇s method. Their synergistic effect were evaluated by isobolographic analysis and allowed the determination of benefit indices （BI） for respective combinations. The model of convulsive rats kindled by penicillin topically injected into cortex was used to investigated the content of Glu, Asp, Gly and GABA in hippocampus using high performance liquid chromatography （HPLC）. Results Combinations of PTA and UTA at the ratio of 4：1 were synergistic in MES test and antagonistic in acute toxicity test, showing the best profile for combinations of PTA with UTA. In contrast, the ratios of 1 ：4 and 1 ： 1, despite synergistic in MES test, were additive in acute toxicity test. The 4：1 combination and two drugs alone significantly decreased Glu level and increased GABA level in the hippocampus, but the GABA level in the 4：1 combination group was higher than that in the two drugs alone groups. They did not have significant influence on the levels of ASp and Gly. Conclusion Combinations of PTA and UTA at 4：1 ratio demonstrated synergistic effect in anticonvulsant action and antagonistic effect in toxicity. The anticonvulsant mechanism might be related to decreasing the excitability of Glutamatergic neurons and increasing the inhibition of GABAergic neurons.

半夏高产繁殖栽培技术研究

通过不同播种密度及氮肥、磷肥施用量对半夏产量的影响研究,为半夏(Pinellia ternata)高产繁殖栽培技术提供依据。采用3因子5水平二次通用旋转组合设计方法进行试验。结果表明,3个试验因子对半夏块茎产量影响的重要程度从大到小为播种密度、P2O5施用量、N施用量。建立了各因素与半夏块茎产量的数学模型,通过对模型解析和模拟寻优,提出半夏块茎产量≥2 700 g/m2和半夏块茎产量≥3 100 g/m2的优化栽培方案。

Optimization of Inducing Conditions for Loose Callus of Pinellia ternata

[Objective] The research aimed to establish the cell suspension culture system of Pinellia ternate.[Method] The petiole of Pinellia ternate was as the explant.The orthogonal test was used to study the influences of four plant growth substances（2,4-D,NAA,picloram and KT）and their mixture ratios on the formation of loose callus.[Result] The induction effect of 2,4-D and picloram on the petiole callus of Pinellia ternate was the most significant.Then,the second ones were KT and NAA.The optimal medium which induced Pinellia ternate petiole to form the loose callus was MS＋0.5 mg/L of 2,4-D＋1.0 mg/L of NAA＋1.0 mg/L of picloram＋1.5 mg/L of KT.[Conclusion] The research laid the foundation for extracting the active ingredient from the cell suspension of Pinellia ternate and producing the artificial seed.

西和半夏凝集素抗虫基因克隆及对棉花的遗传转化

采用同源序列克隆方法,通过设计特异引物克隆甘肃西和半夏(Pinellia ternata)凝集素基因pta,其序列全长l069bp,编码区804bp,编码268个氨基酸残基,有3个典型的甘露糖结合位点,预测分子量和等电点分别为29.1kD和7.77,GenBank登录号AY725425。构建了35S启动子驱动的植物表达载体pBI-pta。建立了陇棉2号花粉管通道法遗传转化技术,获得5株卡那霉素抗性和PCR检测阳性植株,为建立棉花转基因方法和培育抗虫种质奠定了基础。

不同施肥方式下半夏重金属污染评价

对施专用肥、习惯种植(草木灰+普钙)、不施肥3种处理的半夏(Pinellia ternata)块茎烘干后,按照直径d>1.5 cm、1.0 cm≤d≤1.5 cm、d<1.0 cm进行分级,测定半夏中铅(Pb)、镉(Cd)、砷(As)、铜(Cu)的含量,并采用单项与综合污染指数法对半夏中重金属污染进行评价。结果表明,施专用肥与习惯种植两种处理对半夏中重金属含量的影响相当,不施肥处理的影响较小。半夏中重金属的平均污染指数从大到小为Cd、Cu、Pb、As,综合污染指数未达污染级别。3种处理得到的半夏,中等大小(1.0 cm≤d≤1.5 cm)的块茎各重金属含量、平均污染指数、综合污染指数均达到安全标准,属于安全级别。施专用肥的半夏块茎4种重金属平均含量较低,是3种处理中最优的施肥方式。Cd为影响半夏清洁度的主要因子,在专用肥料的推广中,应注重对土壤中Cd的控制。

不同土壤基质配比对半夏生长和品质的影响

试验使用园土、沙土和腐殖土按照一定的比例混配,分析不同土壤基质配比对半夏生长和品质的影响。结果表明,处理B(V_(园土)∶V_(沙土)∶V_(腐殖土)=4∶3∶3)效果最好,各农艺性状和药效成分含量均显著高于其他3个处理。

不同地区半夏无机元素含量分析

测定了6个半夏居群及其土壤样品中的6种人体必需元素,以了解半夏对元素的富集能力和分布规律,为其品质评价提供依据。结果显示:不同居群半夏及其土壤的元素含量存在着显著性差异;同一居群的半夏和土壤元素含量不具有相关性;半夏皮中元素含量最高;药材元素含量相关性显示Zn和Se、Fe和N i、Fe和Ca具有协同吸收特点;富集系数表明半夏对元素富集能力较低。

配伍对大黄致泻作用的影响

中医临床应用大黄,常与若干种其它中药以不同比例配伍,组成相对固定的组对(药对、对药或对子),形成多种方剂的核心.中医药著作中,对大黄配伍的讨论甚多,但现代实验尚少报道。材料一、药材大黄为1983年自青海省药材公司购进的散装箱吉,基源为掌叶大黄Rhe-um palmatum L.或唐古特大黄R.tangu-ticum Maxim ex Batf。人参为1989年自北京市药材公司购进的二级生晒参。半夏为1983年自北京市药材公司购进的姜半夏,基源为半夏Pinellia ternata(Thunb.)Bre-it.,同时购进的有粉甘草(甘草Glyrrhizasp.)、黑附片(附子Aconitum carmich-

半夏愈伤组织诱导与分化最适培养基筛选

以半夏[Pinellia ternata(Thunb.)Breit.]叶片为试验材料,研究不同培养基配方对半夏叶片的诱导脱分化、愈伤组织增殖及分化的影响。结果表明,适合半夏叶片诱导脱分化的培养基为MS+0.8 mg/L6-BA+1.8 mg/L NAA+0.1 mg/L 2,4-D;适合愈伤组织增殖的培养基为MS+0.5 mg/L 6-BA+1.0 mg/L2,4-D;适合分化生长的培养基为MS+0.8 mg/L 6-BA+0.2 mg/L NAA+3.0 mg/L 2,4-D。

Research on "Erchen" in Erchen decoction

Traditional Chinese Medicine has a long history of aging medicines,with the Erchen decoction(it is a traditional Chinese botanical formula),produced by Chenpi(Citri Reticulatae Pericarpium)(CRP)and Banxia(Pinelliae Rhizoma)(PR),as having the most research value.While modern pharmacological studies have found that the main active components of CRP are flavonoids and volatile oils.The relative contents of 30 CRP flavonoids increase with aging time.However,the expectorant effect of CRP is not proportional to its aging time,and its volatile oil composition is thought to be negatively correlated with its aging time,yet this correlation has not yet been established by research.In addition,different experimental measurement methods have found that aging has a reduced effect on the toxic components of different processing types of PR,such as calcium oxalate needle crystals and protein substances.These research also noted that different processing methods also resulted in differences in the efficacy of PR.For example,during the aging process of Jiangbanxia(Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine)and auxiliary materials,6-gingerol was produced,which enhanced the anti-vomiting effects of PR but weakened its expectorant and cough function.Using a method of Alum processed PR,increases the contents of Inosine,Guanosine,Adenosine and Succinate,which thus enhances its expectorant effect.Finally,the contents of 6-Gingerol,Glycyrrhizin,Ammonium Glycyrrhizate,and other substances produced in the aging process of PR,can be used as base compounds for producing additional products of PR.The research on the efficacy and uses of Erchen decoction,particularly compounds made with CRP and PR is voluminous,but no standards currently exist for effective and standardized application of these substances.This paper reviews the research with an aim to establish standardization of methods and of compound aging time.

外源NO对高温胁迫下半夏抗氧化系统的影响

研究了外源NO供体(SNP)对高温胁迫下半夏抗氧化酶及相关生理指标的影响。结果表明,100、200μmol/L SNP提高了半夏叶片中超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性,增加了游离脯氨酸(Pro)、可溶性糖、可溶性蛋白含量,降低了丙二醛(MDA)含量。由此可见,适宜浓度的SNP可提高半夏抗氧化能力,缓解高温胁迫造成的伤害。

荆半夏多倍体新种质诱导研究

设置不同浓度和不同诱导时间的秋水仙素,采用混培法和覆棉法对荆半夏[Pinellia. ternata(Thumb) Berit]进行离体和活体的多倍体诱导,以期建立荆半夏多倍体的化学诱变技术体系。结果表明:采用混培法对荆半夏进行离体多倍体诱导的最佳条件是0.1%秋水仙素溶液处理3d,成活率为48.7%,诱导率28.53%;采用覆棉法进行活体多倍体诱导的最佳条件是0.1%秋水仙素溶液处理6d,诱导率为60.91%,成活率为31.83%;经倍性鉴定,获得了荆半夏多倍体植株(2n=16x=208),其多倍体植株叶片大而肥厚、叶绿素含量增加,植株紧凑、粗壮,且叶片的气孔变大、密度减小。