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.

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.

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.

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。

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.

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。

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.

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.

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.

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

Objective： To evaluate the regulatory effects of Banxia （Pinellia ternata, P） and Huanglian （Coptis chinensis, C） drugpair, derived from Banxiaxiexin soup, on the gastrointestinal movement of mice with functional dyspepsia. Methods：Mice were treated with different proportions of the P and C drug pair （1：1, 3：1, and 4：1） for 10 days, and subsequentlyinjected with atropine （ATR） or neostigmine （NEO）. The effects of the different proportions of P and C were evaluatedbased on the alvine advance rate. In addition, we used the same modeling method used in the first experiment andadministered P： C at ratio of 3：1 and at different doses respectively （4.68 g/L, 2.34 g/L, and 1.17 g/L）, and tested levelsof the gastrointestinal hormones, gastrin （GAS）, vasoactive intestinal polypeptide （VIP）, and somatostatin （SS） in thesmall intestinal tissue using an enzyme-linked immunosorbent assay. Results： In the groups of NEO-induced mice, P：Cat ratios of 1：1, 3：1, and 4：1 significantly reduced the alvine advance rate compared with the NEO model group （P =0.003, P = 0.012 and P = 0.021, respectively）. In the groups of ATR-induced mice, only P：1 at ratio of 3：1 significantlyincreased the alvine advance rate compared with the ATR model group （P = 0.007）. After exposure to P： C at ratio of 3：1and at different dose, the GAS level was lower in the low-, medium-, and high-dose NEO groups than that in the NEOmodel group （P = 0.001, P = 0.004, and P = 0.003, respectively）. The VIP levels were higher in the medium-andhigh-dose NEO groups than that in the NEO model group （P = 0.004 and P = 0.002, respectively）. In addition, the SSlevel increased in the NEO medium-dose group compared with that in the NEO model group （P = 0.002）. The GAS levelwas higher in the ATR medium- and high-dose groups than in the ATR model group （P = 0.007 and P = 0.021,respectively）. The VIP level was lower in the ATR low-, medium-, and high-dose than that in the ATR model group （P =0.001, P = 0.001, and P = 0.001, respectively）. Furthermore, the SS level was lower in the ATR medium- and high-dosegroups than that in the ATR model group （P = 0.001 and P = 0.006）. Conclusion： The PC drug pair bidirectionallyadjusted the NEO- and ATR-induced functional dyspepsia in mice by modulating GAS, VIP, and SS levels in theintestine.

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.

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.

The effects of pinellia total alkaloids on the amino acid concentration and the GABAA receptor expression in hippocampus region of epileptic rats

The antiepileptic effect of pinellia total alkaloids（PTA） on penicillin（PNC） chronically kindled rats was investigated. We investigated the effects of PTA on Glu,Asp,Gly andγ-aminobutyric acid（GABA） concentrations and the expression level of cerebral GABA_A receptor in hippocampus.The influence of PTA on epilepsy seizure latency and degree in PNC chronically kindled rats were observed.High performance liquid chromatography（HPLC） was adopted to measure the concentrations of Glu, Asp,Gly and GABA in hippocampus. Reverse transcriptase-polymerase chain reaction （RT-PCR） was used to determine the expression of cerebral GABAA receptor mRNA. Compared with normal rats, the levels of GABA and Gly decreased obviously while the level of Glu and Asp increased significantly in model rats. The cerebral GABAA receptor mRNA level was also decreased at the same time. The difference was statistically different compared to the control group. PTA could prolong the latent period of the penicillin induced seizure and weaken the extent of seizure, compared with the model group without PTA treatment. Moreover, PTA increased the level of GABA and the expression level of GABAA receptor, while decreased the level of Glu significantly. However, it had no obvious effect on the level of Gly and Asp. Pre-treatment of PTA can also increase the GABAA receptor mRNA level. In conclusion, PTA could alleviate the PNC chronically kindled rat seizure. It increased the GABA level and the expression of GABAA receptor, and it decreased the Glu concentration.

Preliminary analysis of the protein contents in Pinellia ternata with different types of leaves

Total proteins extracted from Pinellia ternata with different types of leaves have been assayed by UV absorbance, blue native polyacrylamide gel electrophoresis （BN-PAGE） and sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS- PAGE）. We found that the total protein contents of P. ternata with the herbaceous peony and claw types of leaves were much higher than that with other types of leaves. P. ternata collected from Nanchuan County, except the ones with herbaceous peony leaves, displayed a high similarity with each other. Both UV absorbance and cluster analysis indicated that the protein content in P. ternata with the herbaceous peony leaves was less affected by habitats than that with other types of leaves. The results showed that it was necessary to increase the homogeneity of seeds for improving the protein content ofP. ternata.

三叶半夏(Pinellia ternata(Thunb.)Breit)的间歇浸没培养

目的:利用间歇浸没培养反应器对药用植物三叶半夏(Pinellia ternata(Thunb.)Breit)进行组培快繁研究。方法:以增殖倍数、鲜重、平均鲜重、折干率和组培苗状态为指标,一方面对三叶半夏叶片、叶柄及丛生芽体进行培养,考察适合间歇浸没培养的外植体类型;另一方面以叶柄为外植体对三叶半夏进行间歇浸没培养、固体培养和液体摇瓶培养,考察三种培养方式对其生长的影响。结果:不同的外植体在反应器内培养时,从增殖情况来看,丛生芽作为外植体得到的增殖倍数高达39.15,叶柄为27.65,叶片为18.05。鲜重方面与增殖倍数具有相同的规律。从平均鲜重和折干率来看,无显著差异。由组培苗形态来看,以叶柄诱导的组培苗最为健壮,而以丛生芽诱导的组培苗则较为细弱。不同培养方式下的增殖倍数分别为:间歇浸没培养方式最高为24.73,其次固体培养为14.75,液体摇瓶培养为12.26,差异显著。平均鲜重和折干率均为间歇浸没培养最优。由组培苗形态来看固体培养优于间歇浸没培养,反应器得到的组培苗较为细弱。在保卫细胞长宽和叶绿体个数上两者没有显著性差异。而液体摇瓶培养玻璃化导致叶绿体个数减少,不适合培养三叶半夏的。结论:较适合间歇浸没培养的外植体类型为叶柄和丛生芽,以叶柄为优。间歇浸没培养方式相对于传统的固体和液体培养在各项生理指标上表现较好,具有大规模培养三叶半夏的潜力。

Three new alkaloids isolated from the stem tuber of Pinellia pedatisecta

The present study was designed to determine the chemical constituents of the stem tuber of Pinellia pedatisecta. The chemical constituents were isolated and purified by various chromatographic techniques, and their structures were elucidated on the basis of physicochemical properties and spectral data. Three new alkaloids(compounds 1, 2, and 3) were obtained and identified as 9-((5-methoxypyridin-2-yl)methyl)-9 H-purin-6-amine(1), 4-(2-(2, 5-dioxopyrrolidin-1-yl)ethyl)phenyl acetate(2), and N-(9-((5-methoxypyridin-2-yl)methyl)-9 H-purin-6-yl)acetamide(3). These compounds were evaluated for their cytotoxicity against human cervical cancer He La cells. Compounds 1 and 3 significantly inhibited the proliferation of He La cells with IC_(50) values being 3.02 ± 0.54 and 7.16 ± 0.62 μmol·L^(-1), respectively.