Age Discrimination Based on Volatile Components of Arisaema Cum Bile / 中国实验方剂学杂志

ObjectiveTo discriminate the age of Arisaema Cum Bile， the combination of headspace solid-phase microextraction （HS-SPME） with gas chromatography-mass spectrometry （GC-MS） was applied to explore the differences of volatile components of unfermented， 1-year fermented， 2-year fermented， and 3-year fermented Arisaema Cum Bile. MethodSamples with different fermentation durations were collected and HS-SPME-GC-MS technology was employed to detect the volatile components of each sample. The relative contents of detected volatile components were processed and analyzed by chemometrics methods such as principal component analysis （PCA）， hierarchical cluster analysis （HCA）， and partial least squares discriminant analysis （PLS-DA）. ResultThe results showed that 145 volatile components were identified. Among these volatile components， the relative contents of heterocyclic， alcohols， aldehydes and aromatics were high. PCA， HCA， and PLS-DA can effectively separate Arisaema Cum Bile with four different ages. Based on variable importance in projection （VIP） value > 1， 73 markers of differential volatile components were identified. The content of 2，6，11-trimethyldodecane and m-xylene in unfermented samples was the highest， and the content difference between them and those in fermented samples was significant （P<0.05）. 2，3-butanediol was detected only in 1-year samples， octane was detected only in 2-year samples， and ethyl heptanoate was detected only in 3-year samples. These components can be used as odor markers for Arisaema Cum Bile with different fermentation years. ConclusionThe identification method of volatile components of Arisaema Cum Bile was established by HS-SPME-GC-MS technology， which can realize the rapid identification of unfermented， 1-year fermented， 2-year fermented， and 3-year fermented samples， and provide a scientific basis for the standardization of processing technology and quality standards of Arisaema Cum Bile.

Investigation on the protective effect of Arisaema Cum Bile on MPTP-induced Parkinson’s disease model mice based on PKA signaling pathway / 中国药房

OBJECTIVE To investigate the improvement effects of Arisaema Cum Bile on Parkinson’s disease （PD） model mice and its potential mechanism. METHODS Sixty male C57BL/6J mice were randomly divided into normal group， model group， Arisaema Cum Bile low-dose group [0.39 g/（kg·d）]， Arisaema Cum Bile high-dose group [1.56 g/（kg·d）] and positive control drug Levodopa tablet group [80 mg/（kg·d）]， with 12 mice in each group. Except that normal group was given constant volume of normal saline， other groups were given 1-methyl-4-phenyl-1，2，3，6-tetrahydropyridine [MPTP，35 mg/（kg·d）] intraperitoneally for 5 consecutive days to induce subacute PD model； after modeling， they were given relevant medicine continuously for 7 d； rod climbing test and line suspension test were performed 1 d before modeling， on the 5th day of modeling and after the last medication. The number of tyrosine hydroxylase （TH）-positive neurons in the substantia nigra of mice were measured by immunofluorescence； the levels of interleukin 1β （IL-1β） and tumor necrosis factor α（ TNF-α） in serum and the levels of IL- E-mail：qhwang668@sina.com 1β， TNF-α， cyclooxygenase-2 （COX-2） and inducible nitric oxide synthase （iNOS） in the substantia nigra of mice were measured by enzyme-linked immunosorbent assay. The expression levels of cAMP-dependent protein kinase catalytic subunit α （PKA C-α）， glutathione peroxidase 4 （GPX4） and ferritin heavy chain polypeptide 1 （FTH1） proteins in the substantia nigra of mice was measured by Western blot. RESULTS After last medicine， compared with the normal group， mice in the model group had significantly longer pole-climbing time （P＜0.01）， significantly lower line suspension scores （P＜0.01）， significantly fewer TH-positive neurons in the substantia nigra （P＜0.01）， significantly higher serum concentrations of IL-1β and TNF-α and nigrostriatal concentrations of IL-1β， TNF-α， COX-2 and iNOS （P＜0.01）， while lower protein expression levels of GPX4， PKA C-α and FTH1 in the substantia nigra （P＜0.05 or P＜0.01）. Compared with the model group， the above indexes of mice were significantly returned in Arisaema Cum Bile high-dose group （P＜0.05 or P＜ 0.01）. CONCLUSIONS Arisaema Cum Bile can improve motor impairment and reduce apoptosis of nigrostriatal TH neurons in MPTP-induced PD mice， and has neuroprotective effects on model mice； this may be related to its inhibition of neuroinflammation and the inhibition of ferroptosis by up-regulating PKA signaling pathway.

Analysis of Q-markers of Arisaema Cum Bile Acting on Stroke Based on UPLC-QTOF-MS/MS and TCMIP v2.0 / 中国实验方剂学杂志

ObjectiveTo predict the possible quality markers （Q-markers） of Arisaema Cum Bile in the prevention and treatment of stroke based on ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spetrometry （UPLC-QTOF-MS/MS） and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP） v2.0. MethodUPLC-QTOF-MS/MS was employed with the mobile phase of 0.1% formic acid aqueous solution （A）-0.1% formic acid acetonitrile solution （B） for gradient elution （0-3 min， 0.2%-5%B； 3-5 min， 5%-8%B； 5-8 min， 8%-10%B； 8-14 min， 10%-25%B； 14-18 min， 25%-50%B； 18-20 min， 50%-70%B； 20-21 min， 70%-98%B； 21-23 min， 98%B； 23-24 min， 98%-0.2%B； 24-26 min， 0.2%B）， the flow rate of 0.5 mL·min-1 and electrospray ionization （ESI）. High quality MS/MS data were scanned in positive and negative ion modes with scanning range of m/z 50-1 500. A local database of the chemical constituents in Arisaema Cum Bile was established by UNIFI 1.8. Then the chemical constituents in Arisaema Cum Bile were characterized by matching with the local database and comparing with the reference substances and literature information. TCMIP v2.0 was used to obtain the targets corresponding to the identified components of Arisaema Cum Bile and stroke， and the "disease-formula" correlation analysis was carried out to screen the core targets by topological eigenvalues. DAVID 6.8 was used for enrichment analysis of Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway of core targets. According to the "five principles" of Q-markers and combined with literature reports， the Q-markers of Arisaema Cum Bile in the prevention and treatment of stroke were predicted， and the core components acting on these target genes were obtained. Cytoscape 3.8.0 was employed to draw the network diagram of "medicinal materials-active ingredients-target genes-pathways". Finally， AutoDock Vina 1.2.2 was used to calculate and verify the molecular docking between the candidate components and the key targets. ResultA total of 76 chemical components was identified in positive and negative ion modes， 85 core targets were collected for Arisaema Cum Bile in the prevention and treatment of stroke. A total of 31 stroke-related pathways， 23 target genes and 9 main active components of Arisaema Cum Bile acting on these genes were screened， and then we determined 4 possible Q-markers for Arisaema Cum Bile in the prevention and treatment of stroke according to the "five principles". ConclusionThe possible Q-markers of Arisaema Cum Bile for stroke are gallic acid， apigenin-6，8-di-C-glucoside， apigenin and cholic acid， and the target of these four components may be estrogen receptor alpha （ESR1）.

Isolation and Identification of Arisaema Cum Bile Fermentation Strain and Preliminary Investigation on Its Compound Strain Fermentation / 中国实验方剂学杂志

ObjectiveIn order to solve the problem that the quality and stability of Arisaema Cum Bile in the fermentation process with hybrid bacteria were not easy to control， the microorganism in the fermentation process of Arisaema Cum Bile was isolated and identified， the dominant strains were screened and the fermentation process of Arisaema Cum Bile with compound bacteria was investigated. MethodThe submerged culture during the fermentation process of Arisaema Cum Bile was taken out for strain separation and purification. Bacteria and fungi multiphase identification and detection methods and automatic microbial analysis system were used to analyze and compare DNA sequences and identify microorganisms. The isolated and identified strains were respectively inoculated and fermented. After screening the dominant strains， a preliminary exploration of compound strain fermentation were carried out. The contents of index components in Arisaema Cum Bile fermented by compound strain and traditional Arisaema Cum Bile were compared by ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry （UPLC-QqQ-MS/MS）. Mmobile phase was 0.1% formic acid acetonitrile solution （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-2 min， 35%-45%A； 2-10 min， 45%-48%A； 10-12 min， 48%-100%A； 12-12.01 min， 100%-35%A； 12.01-15 min， 35%-65%A）， the flow rate was set at 0.35 mL·min-1. The mass spectrographic analysis employed electrospray ionization （ESI）， negative ion acquisition mode and multiple reaction monitoring （MRM） scanning mode were adopted to collect information， the collection range was m/z 50-1 000. ResultEight microorganisms were isolated and identified from the submerged culture of Arisaema Cum Bile. Among them， Enterococcus sp. （anaerobic） and E. casseliflavus were selected as the dominant strains in the fermentation process. Compared with the traditional fermentation method， the contents of chenodeoxycholic acid， hyodeoxycholic acid and hyocholic acid in free cholic acid increased by 1.76， 0.06， 0.19 mg·g-1， respectively. In bound cholic acid， glycochenodeoxycholic acid， taurochenodeoxycholic acid， glycohyodeoxycholic acid， taurohyodeoxycholic acid， glycohyocholic acid， taurine porcine cholic acid decreased by 0.63， 0.23， 0.26， 0.16， 0.03， 0.04 mg·g-1， respectively. ConclusionArisaema Cum Bile with compound strain fermentation （Enterococcus sp. and E. casseliflavus） can be fermented more completely， the fermentation cycle can be shortened， and the quality and stability of products can be improved.

Research progress on chemical constituents and biological activities of Arisaematis Rhizoma / 中国中药杂志

Arisaematis Rhizoma included in the Chinese Pharmacopoeia is the dried tuber of Arisaema erubescens, A. heterophyllum or A. amurense in the family Araceae. This paper mainly focuses on the classification and summary of the chemical components and structures reported in recent years in the above three varieties of this medicinal material included in the pharmacopoeia, including alkaloids, flavonoids, phenylpropanoids, lignans and benzene ring derivatives, steroids and terpenes, glycosides and esters, etc. Then we reviewed the reported biological activities of these chemical components, including cytotoxicity, antitumor activity, antibacterial activity, nematicidal activity, etc. Although there have been reports on the review of the chemical composition of the medicinal material, the structure and classification of the chemical composition in these reviews are not clear enough. This review provides a basis for the later study of the chemical composition of this medicinal material, especially the identification of the chemical structures. And most of the current reviews on the biological activity of this medicinal material are mainly for the crude extract. This paper mainly summarized the biological activity of related monomer compounds and expected to lay a foundation for the development of novel high-efficiency and low-toxicity active leading compounds from Arisaematis Rhizoma.

Effect of Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma on Water Metabolism and Hemorheology in Rats / 中国实验方剂学杂志

Objective:To discuss the effect of Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma on water metabolism and hemorheolog in rats. Method:A total of 50 healthy SD rats were randomly divided into 5 groups, namely normal group, high-dose Arisaema Rhizoma group (1.2 g·kg·d-1), low-dose Arisaema Rhizoma group (1.2 g·kg·d-1), high-dose Pinelliae Pedatisectae Rhizoma group (1.2 g·kg·d-1) and low dose Pinelliae Pedatisectae Rhizoma group (0.3 g·kg·d-1), with 10 rats in each group. The effects of Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma on water metabolism and hemorheology were compared, with the micturition index, fecal volume, fecal water content, pathological morphology of submandibular gland, aquaporin (AQP2) content in kidney, adenosine cyclic 3′,5′-(hydrogen phosphate)/current good manufacture practices (cAMP/cGMP) and Na+-K+-ATP enzyme content in serum as the evaluation indexes of water metabolism, and the whole blood viscosity (WBV) and erythrocyte aggregation index (RE) as the evaluation indexes of hemorheolog. Result:Compared with normal group, the fecal volume, fecal water content and AQP2 expression decreased significantly， acinus of submandibular gland were atrophied， micturition index, cAMP/cGMP and Na+-K+-ATP enzyme content in serum were increased significantly， WBV and RE were increased significantly in high-dose Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma groups（P<0.05）, but with no significant difference between them. Conclusion:High-dose Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma groups had significant effects on water metabolism and hemorheology in rats, suggesting that attention shall be given to the safety of Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma. And this study provides a theoretical basis for the rational use of the two traditional Chinese medicines in clinic.

Investigation of Protective Effect of Different Bile and Their Arisaema cum Bile on Acute Lung Injury Rats Induced by LPS / 中国实验方剂学杂志

<b>Objective::To compare the protective effect of different bile (porcine bile, oxgall and sheep bile) and their Arisaema cum Bile on rats with acute lung injury, so as to provide reference for the selection of bile and the classification of decoction pieces of Arisaema cum Bile. <b>Method::Wistar male rats were randomly divided into 8 groups (<italic>n</italic>=12), including blank group, model group, porcine bile group, oxgall group, sheep bile group, Arisaema cum Bile with porcine bile group, Arisaema cum Bile with oxgall group and Arisaema cum Bile with sheep bile group. Rats in each treatment group were given corresponding drug solution by gavage at 2.52 g·kg^-1 every day, and rats in the model group and the blank group were given the same volume of normal saline by gavage every day for a total of 8 days. On the 8^th day, after 1 h of administration, rats in the model group and each treatment group were intraperitoneally injected lipopolysaccharide (LPS, 2 mg·kg^-1) to prepare rat lung injury model. Blood and lung tissues were collected from every four rats at 3, 6, 24 h after LPS injection, respectively. Lung coefficient, lung water content and wet weight/dry weight ratio of lung tissue (<italic>W</italic>/<italic>D</italic>) were measured, and the levels of tumor necrosis factor (TNF)-<italic>α</italic>, interleukin (IL)-6 and thromboxane B₂ (TXB₂) in serum and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and the contents of malondialdehyde (MDA) and matrix metalloproteinase-9 (MMP-9) in lung tissue were determined by enzyme linked immunosorbent assay (ELISA). The pathological morphology of rat lung tissue was observed and the score of lung tissue injury was calculated. <b>Result::Compared with the model group at the same time point, the lung coefficient, <italic>W</italic>/<italic>D,</italic> lung water content, contents of TNF-<italic>α</italic>, IL-6 and TXB₂ in serum, contents of MDA and MMP-9 in lung tissue of rats in each treatment group were all decreased, and most of them had significant differences (<italic>P</italic><0.05, <italic>P</italic><0.01), but the activities of GSH-Px and SOD were all increased, and most of them had significant differences (<italic>P</italic><0.05, <italic>P</italic><0.01). The oxgall group and the sheep bile group were superior to the porcine bile group in most of the indexes, the Arisaema cum Bile with oxgall group and the Arisaema cum Bile with sheep bile group were superior to the Arisaema cum Bile with porcine bile group, and some of them had significant differences(<italic>P</italic><0.05, <italic>P</italic><0.01). <b>Conclusion::Each bile group and each Arisaema cum Bile group all have protective effects on rats with acute lung injury induced by LPS, and the therapeutic effect of oxgall and sheep bile is better than that of porcine bile, the therapeutic effect of Arisaema cum Bile prepared by oxgall and sheep bile is better than that of Arisaema cum Bile prepared by porcine bile.

Comparative study on HPLC characteristic chromatogram of water extracts of Arisaematis Rhizoma and its processed products / 中草药

Objective: To establish and analyze the HPLC characteristic chromatogram for water extracts of Arisaematis Rhizoma (AR) and its processed products, Arisaematis Rhizoma Preparatum (ARP) and Arisaema Cum Bile (ACB). The research provided reliable method and scientific basis for their quality control. Methods: The separation was performed on an Agilent C18 (200 mm × 4.6 mm, 5 μm) column with gradient elution of 0.2% acetic acid water and 0.2% acetic acid acetonitrile. The similarity was analyzed with software “Similarity Evaluation System for Chromatographic Fingerprint of TCMs (Version 2012)”. The cluster analysis was performed by SPSS 23.0. The principal component analysis was performed by SIMCA 14.1. Results: HPLC characteristic chromatogram for water extracts of AR, ARP, and ACB were established. There were 19, 20 and 13 common peaks in AR, ARP and ACB, respectively. A total of eight characteristic peaks were identified as F1 (xanthine), F3 (uracil), F4 (hypoxanthine), F5 (uridine), F6 (guanosine), F7 (adenosine), F14 (schaftoside), and F16 (isoschaftoside), respectively. The intragroup similarities of AR, ARP, and ACB were all above 0.8 and each was clustered into one type, and the intergroup similarities among AR and its processed products were all below 0.4. The main components of AR were F16 (isoschaftoside), F14 (schaftoside), F17and F15. The main components of ARP were F16 (isoschaftoside) and F1 (xanthine). The main components of ACB were F3 (uracil), F2, F5 (uridine) and F1 (xanthine). Conclusion: The method can effectively identify AR, ARP and ACB, and provide a scientific basis for their quality control.

Cloning and characterization of chalcone synthase and chalcone isomerase genes in Arisaema heterophyllum / 中国中药杂志

Chalcone synthase( CHS) and chalcone isomerase( CHI) are key enzymes in the biosynthesis pathway of flavonoids. In this study,unigenes for CHS and CHI were screened from the transcriptome database of Arisaema heterophyllum. The open reading frame( ORFs) of chalcone synthase( Ah CHS) and chalcone isomerase( Ah CHI) were cloned from the plant by RT-PCR. The physicochemical properties,expression and structure characteristics of the encoded proteins Ah CHS and Ah CHI were analyzed. The ORFs of Ah CHS and Ah CHI were 1 176,630 bp in length and encoded 392,209 amino acids,respectively. Ah CHS functioned as a symmetric homodimer. The N-terminal helix of one monomer entwined with the corresponding helix of another monomer. Each CHS monomer consisted of two structural domains. In particular,four conserved residues define the active site. The tertiary structure of Ah CHI revealed a novel open-faced β-sandwich fold. A large β-sheet( β4-β11) and a layer of α-helices( α1-α7) comprised the core structure. The residues spanning β4,β5,α4,and α6 in the three-dimensional structure were conserved among CHIs from different species. Notably,these structural elements formed the active site on the protein surface,and the topology of the active-site cleft defined the stereochemistry of the cyclization reaction. The homology comparison showed that Ah CHS had the highest similarity to the CHS of Anthurium andraeanum,while Ah CHI had the highest similarity to the CHI of Paeonia delavayi. This study provided the basis for the functional study of Ah CHS and Ah CHI and the further study on plant flavonoid biosynthesis pathway.

Study of medicinal properties of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile based on delayed luminescence of Scenedesmus sp / 中草药

Objective To find delayed luminescence parameters that could characterize the cold and hot properties of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile. Methods Delayed luminescence of Arisaematis Rhizoma Preparatum with addition of Scenedesmus sp. within 26 d after decoction was measured in unequal interval, with aim to verify the stability of the natural delayed luminescence average strength and the linear fitting slope value (k) of excitation delayed luminescence. The delayed luminescence of six batches of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile was measured using biological indicator method, and the content of β-sitosterol in Arisaematis Rhizoma Preparatum and β-sitosterol, bilirubin, and cholic acid of Arisaema Cum Bile was determined using high performance liquid chromatograph (HPLC) to analyze the correlation of k value and the above components content of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile. Results K value of excitation delayed luminescence within 14 d after decoction was steadier than natural delayed luminescence average strength, and k values of six batches of Arisaematis Rhizoma Preparatum were all higher than that of Arisaema Cum Bile. A significant negative correlation between β-sitosterol contents and k values of six batches of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile was found, and no significant negative correlation between bilirubin and cholic acid contents and k values of Arisaema Cum Bile was found. Conclusion K value of excitation delayed luminescence could indicate the differences of medicinal properties of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile, which provides a new method for the study of medicinal properties of Chinese materia medica.

Effect of fermentation on components of bile acids in Arisaema Cum Bile and determination of three kinds of free bile acids in Arisaema Cum Bile / 中国中药杂志

The aim of this study is to analyze the compositions of main bile acids in fermented and mixed processing products of arisame cum bile from pig bile, and to establish a method for content determination of bile acids in fermented Arisaema Cum Bile. Fermented and mixed processing products were prepared from arisaematis rhizome and arisaematis rhizoma preparatum with pig bile respectively. Then the differences in bile acids compositions between such two kinds of products were compared by high performance liquid chromatography and evaporative light-scattering detector (HPLC-ELSD). With three kinds of free bile acid compositions as the indicators, HPLC-ELSD method was adopted to determine the content of bile acid compositions in fermented product,on Agilent Eclipse XDB C₁₈(4.6 mm×250 mm, 5 μm) chromatographic column, with acetonitrile and 0.1% glacial acetic acid solution (55:45) as mobile phase, at a flow rate of 1 mL·min⁻¹, column temperature of 30 °C, drift tube temperature of 90 °C, and a nitrogen flow rate of 2.2 mL·min⁻¹. The results showed that the bile acids in fermented bile Arisaema were mainly in a free form, while in mixed processing product, the compositions were mainly in a conjugated form. Three kinds of free bile acids, namely porcine cholic acid (HCA), porcine deoxycholic acid (HDCA) and chenodeoxycholic acid (CDCA) in fermented product, showed a good linear relationship in the range of quantification. The average recovery rate was 95.99%-104.3%, complying with the requirements. The results showed that the conjugated bile acids could be transformed into free bile acids during the fermentation of arisaema cum bile. This established method can effectively control the content of bile acids compositions in fermenting arisaema cum bile.

Cerebrosides isolated from Arisaema flavum / 中国中药杂志

Silica gel, Sephadex LH-20, and reverse phase (C-18) column chromatography were used for the research of chemical constituents occurred in Arisaema flavum(Forsk.) Schott. The structures were elucidated by comparison physico-chemical properties and NMR spectroscopic data with those of known compounds. Seventeen cerebrosides were identified as 1-O-β-D-glucopyranosyl-(2S, 3R, 4E, 8E)-2-[(2'(R)-acetoxyoctadecanoyl)amido]-4, 8-octadecadiene-1, 3-diol (1), 2'-O-acetylsoyacerebroside I (2), 1-O-(β-D-glucopyranosyl)-(2S, 3R, 4E, 13Z)-2-[(2'R)-2-hydroxytetradecanoylamino]-1, 3-dihydroxy-4, 13-docosadiene (3), (2S, 3R, 4E, 8E)1-(β-D-glucopyranosyl)-3-hydroxy-2-[(R)-2'-hydroxyhexadecanoyl]amino-9-methyl-4, 8-heptadecadiene (4), (2S, 3R, 4E, 8E)1-(β-D-glucopyranosyl)-3-hydroxy-2-[(R)-2'-hydroxyhexadecanoyl]amino-9-methyl-4, 8-octadecadiene (5), (2S, 3R, 4E, 8E)1-(β-D-glucopyranosyl)-3-hydroxy-2-[(R)-2'-hydroxypalmitoyl]amino-9-methyl-4, 8-octadecadiene (6), (2S, 3R, 4E, 8E)1-(β-D-glucopyranosyl)-3-hydroxy-2-[(R)-2'-hydroxyoctadecanoyl]amino-9-methyl-4, 8-octadecadiene (7), 1-O-(β-D-glucopyranosyl)-(2S, 3R, 4E, 8E)-2-[(R)-2'-hydroxytetradecanoylamino]-4, 8-octadecadiene-1, 3-diol (8), 1-O-(β-D-glucopyranosyl)-(2S, 3R, 4E, 8E)-2-[(R)-2'-hydroxypentadecanoylamino]-4, 8-octadecadiene-1, 3-diol (9), 1-O-(β-D-glucopyranosyl)-(2S, 3R, 4E, 8E)-2-[(R)-2'-hydroxyhexadecanoylamino]-4, 8-octadecadiene-1, 3-diol (10), 1-O-(β-D-glucopyranosyl)-(2S, 3R, 4E, 8Z)-2-[(R)-2'-hydroxyhexadecanoylamino]-4, 8-octadecadiene-1, 3-diol (11), 1-O-(β-D-glucopyranosyl)-(2S, 3R, 4E, 8E)-2-[(R)-2'-hydroxyoctadecanoylamino]-1, 3-hydroxy-4, 8-octadecadiene (12), 1-O-(β-D-glucopyranosyl)-(2S, 3R, 4E)-2-[(R)-2'-hydroxytetracosanoylamino]-1, 3-hydroxy-4-hexadecane (13), 1-O-(β-D-glucopyranosyl)-(2S, 3R, 4R, 8Z)-2N-[(2'R)-2'-hydroxytetracosanoyl]-8-(Z)-octadecene-1, 3, 4-triol (14), 1-O-(β-D-glucopyranosyl)-(2S, 3S, 4E, 8E)-2N-[(2'R)-2'-hydroxyhexadecanoyl]-4-(E), 8-(Z)-octadecadiene-1, 3-diol (15), typhoniside A (16), and 1-O-β-D-glucopyranosyl-(2S, 3R, 8E)-2-[(2'R)-2-hydroxypalmitoylamino]-8-octadecene-1, 3-diol (17). Compounds 1 and 2 were isolated from the plant for the first time, while the remained compounds were isolated from the genus Arisaema for the first time.

Agglutinin isolated from Arisema heterophyllum Blume induces apoptosis and autophagy in A549 cells through inhibiting PI3K/Akt pathway and inducing ER stress / 中国天然药物

Arisaema heterophyllum Blume is one of the three medicinal plants known as traditional Chinese medicine Rhizoma Arisaematis (RA). RA has been popularly used to treat patients with convulsions, inflammation, and cancer for a long time. However, the underlying mechanisms for RA effects are still unclear. The present study was designed to determine the cytotoxicity of agglutinin isolated from Arisema heterophyllum Blume (AHA) and explore the possible mechanisms in human non-small-cell lung cancer A549 cells. AHA with purity up to 95% was isolated and purified from Arisaema heterophyllum Blume using hydrophobic interaction chromatography. AHA dose-dependently inhibited the proliferation of A549 cells and induced G phase cell cycle arrest. AHA induced apoptosis by up-regulating pro-apoptotic Bax, decreasing anti-apoptotic Bcl-2, and activating caspase-9 and caspase-3. In A549 cells treated with AHA, the PI3K/Akt pathway was inhibited. Furthermore, AHA induced increase in the levels of ER stress markers such as phosphorylated eukaryotic initiation factor 2α (p-eIF2α), C/EBP-homologous protein (CHOP), inositol-requiring enzyme 1α (IRE1α), and phosphorylated c-Jun NH-terminal kinase (p-JNK). AHA also induced autophagy in A549 cells. Staining of acidic vesicular organelles (AVOs) and increase in the levels of LC3II and ATG7 were observed in AHA-treated cells. These findings suggested that AHA might be one of the active components with anti-cancer effects in Arisaema heterophyllum Blume. In conclusion, cytotoxicity of AHA on cancer cells might be related to its effects on apoptosis and autophagy through inhibition of PI3K/Akt pathway and induction of ER stress.

Chemical constituents of Arisaema rhizomatum / 中国药学杂志

OBJECTIVE: To investigate the chemical constituents of Arisaema rhizomatum.

Anti-insect potential of lectins from Arisaema species towards Bactrocera cucurbitae.

Bactrocera cucurbitae (Coquillett), also known as melon fruit fly, is one of the major insect pests of cucurbits in several parts of Asia, Africa and Pacific. In the present investigation, effect of lectins from two sources i.e. Arisaema intermedium Blume and Arisaema wallichianum Hook f. (Family-Araceae) has been studied on the development of second instar larvae of melon fruit fly. The lectins were incorporated separately in artificial diet at a concentration of 10 to 160 µg ml-1 and fed adlibitum to the second instar larvae. Both the lectins were found to prolong the development period and significantly inhibited the pupation and emergence in a dose dependent manner. Total development period was found to be prolonged by 3.5 and 2.3 days in case of larvae fed on artificial diet containing A. intermedium (AIL) and A. wallichianum (AWL), respectively. LC50 values calculated on the basis of adult emergence came out to be 32.8 and 29 µg ml-1 for AIL and AWL, respectively. Both the lectins tested, were found to increase the activity of esterases as larvae proceeded from 24 to 72 hr of treatment. The activity of acid phosphatase decreased significantly in larvae reared on diet containing LC50 of AIL, while in case of AWL significant decrease was observed only at 72 hr of treatment. Alkaline phosphatase activity decreased significantly on treatment with both of these lectins. These results showed that AIL and AWL have promising anti-insect potential. So, lectin gene/s from either of these species can be cloned and subsequently can be employed to develop transgenics to control melon fruit flies specifically and insect pests in general. This approach could be used as a part of Integrated pest management (IPM) strategies.

Study on Toxicity and Adverse Effect of Different Processed Samples of Arisaema Amurense Maxim / 中成药

Objective: To investigate the effects of different processing methods (steeping, boiling) and different adjuvants (alum, ginger, bile) on toxicity and adverse effect of Arisaema Amurense Maxim. Methods: The determinations of toxicity and adverse effect were performed by tasting of hot and tingle, test of stimulating rabbit eyes and acute toxicity test. Results: The product processed by alum is better than that by ginger in eliminating hot and tingle taste. The product processed by heating is better than that without heating. The test result of stimulating rabbit eyes is similar to that of hot and tingle taste. The result of acute toxicity test indicates that the toxicity of Arisaema Amurense Maxim is low. Conclusions: It is scientific that Arisaema Amurense Maxim. is mostly processed by alum and heat. The clinical application of Arisaema Amurense Maxim. has a certain safety.

Effects of Processing on Contents of Amino Acid and Part of Inorganic Elements in Arisaema Amurense Maxim / 中成药

Objective: To investigate the effects of different processing methods(steeping, boiling) and different adjuvants(alum, ginger, bile) on contents of amino acid and part of inorganic elements in Arisaema amurense Maxim.. Methods: The amino acid was determined by HPLC, and the inorganic elements were determined by AAS. Results: The total content of amino acid was the highest in raw Arisaema amurense Maxim. and the lowest in Arisaema amurense Maxim. processed with bile, wherease it was similar to other processed samples. The inorganic element content changed after it was processed. Conclusion: The long rinse is the key factor of losing amino acid in Arisaema amurense Maxim. after being processed. The content of inorganic element in adjuvants is the key factor of their content changes in Arisaema amurense Maxim. after it is processed.

Study on the processing method of Arisaema Amurense Maxim. by orthogonal design / 中成药

Objective: To investigate the new processing method of Arisaema Amurense Maxim. Methods: Referring to hot and and tingle taste, we have screened out the best condition in processing Arisaema Amurense Maxim. by testing in orthogonal designs, and compared stimulation, extractives and UV of different samples. Results: The new processing method can save half slum according to Ch P. the stimulation of the two samples processed by Ch P & by new method is similar; the water soluble and ethanol soluble extractives and UV of processed samples by new method is better than that by Pharmacopoeia of PR. China. Conclusion: The new method screened out by testing in orthogonal design, can not only eliminate hot and tingle taste, reduce the lose of effective component, but also save a lot of slum.