This article is a summary of the historical background to Isatis tinctoria in France and elsewhere from the 12th to the 21st century. Isatis tinctoria, also called “Guède”, “Pastel” or “Vouède” accordi...This article is a summary of the historical background to Isatis tinctoria in France and elsewhere from the 12th to the 21st century. Isatis tinctoria, also called “Guède”, “Pastel” or “Vouède” according to the French region origin, is a plant originally used to obtain indigo blue dye mainly to produce colored blue fabrics. A local memory is well established in the Amiens region (northern France) and the Toulouse region (south-western France). No global review of its culture and trade over the long term is known. The aim of this article is to summarize the historical background of Isatis tinctoria in France and elsewhere from the twelfth to the twenty-first century.展开更多
[Objective] This study aimed to compare the microstructure between I. in- digotica Fort. and B. cusia （Nees） Bremek. [Method] Microstructure of I. indigotica Fort. and B. cusia （Nees） Bremek. with different source...[Objective] This study aimed to compare the microstructure between I. in- digotica Fort. and B. cusia （Nees） Bremek. [Method] Microstructure of I. indigotica Fort. and B. cusia （Nees） Bremek. with different sources were compared and analyzed by using digital microscopy. [Result] Comparison result of the microstructure shows that reticulate vessels, single starch granules, multiple starch granules and stone cells are mostly observed in I. indigotica Fort.; pitted vessels, parenchyma cells with small pits, single starch granules and cystolith are mostly observed in B. cusia （Nees） Bremek., which indicated that microstructure of L indigotica Fort. and B. cusia （Nees） Bremek. are significantly different. [Conclusion] Microstructure char- acteristics of I. indigotica Fort. and B. cusia （Nees） Bremek. established in this study can be used to evaluate the quality of these two kinds of Chinese herbal medicines, which also provide scientific basis and methods for comprehensively studying the quality standard of /. indigotica Fort. and B. cusia （Nees） Bremek. Key words Isatis indigotica Fort.; Baphicacanthus cusia （Nees） Bremek.; Microstruc- ture; Quality展开更多
This study was to investigate the effects of three exogenous substances on chemical constituents of Isatis indigotica leavesand their efficacy in alleviating drought stress, and explore the methods of applying exogeno...This study was to investigate the effects of three exogenous substances on chemical constituents of Isatis indigotica leavesand their efficacy in alleviating drought stress, and explore the methods of applying exogenous substances to efficient cultivationof Isatis indigotica. Polyethylene glycol (PEG) was used to simulate drought stress to deal with seeds of Isatis indigotica at thegermination stage (concentration: 0, 10%, 15%, and 20%). Simultaneous operation of exogenous growth regulators [microbialinoculum (MI), γ-aminobutyric acid (GABA) and salicylic acid (SA)] and PEG were implemented in seeds of Isatis indigotica.The effects of drought stress and the mitigation of exogenous substances were observed by statistics of seed germination potential,germination rate, hypocotyl length, and radicle length of each treatment. The effects of exogenous substances on the content ofalkaloids, crude protein and free amino acids in the leaves of Isatis indigotica grown in a greenhouse were determined after sprayingexogenous substances on the plants. The differences of germination potential, germination rate, hypocotyl length, and radicle lengthamong 15% PEG stress treatment, 10% PEG stress treatment and the control were significant (P<0.05). According to the predesignedgermination standard, the seeds did not germinate under 20% PEG stress treatment. When the PEG concentration was 15%, the resultsof seed germination potential and germination rate after adding MI were significantly different from those under stress alone (P<0.05).When exposed to 10% PEG stress, the supplementation of GABA led to a notable increase in radicle length of Isatis indigotica seeds,showing significant differences compared to other three treatments. The application of MI and GABA under 15% PEG stress resultedin a significant increase in the radicle and hypocotyl length of Isatis indigotica seeds compared to other two treatments. The contentof the total alkaloids in leaves of Isatis indigotica was significantly increased after spraying GABA. Meanwhile, the contents of crudeprotein and the total free amino acids were kept constant after spraying exogenous substances. Application of MI and GABA couldalleviate drought stress of Isatis indigotica. The content of the total alkaloids in leaves of Isatis indigotica could significantly increaseafter spraying GABA.展开更多
Isatis indigotica Fort.(Ban-Lan-Gen)is an herbal medicine prescribed for influenza treatment.However,its active components and mode of action remain mostly unknown.In the present study,erucic acid was isolated from Is...Isatis indigotica Fort.(Ban-Lan-Gen)is an herbal medicine prescribed for influenza treatment.However,its active components and mode of action remain mostly unknown.In the present study,erucic acid was isolated from Isatis indigotica Fort.,and subsequently its underlying mechanism against influenza A virus(IAV)infection was investigated in vitro and in vivo.Our results demonstrated that erucic acid exhibited broad-spectrum antiviral activity against IAV resulting from reduction of viral polymerase transcription activity.Erucic acid was found to exert inhibitory effects on IAV or viral(v)RNA-induced pro-inflam-matory mediators as well as interferons(IFNs).The molecular mechanism by which erucic acid with antiviral and anti-inflammatory properties was attributed to inactivation of NF-kB and p38 MAPK signaling.Furthermore,the NF-kB and p38 MAPK inhibitory effect of erucic acid led to diminishing the transcriptional activity of interferon-stimulated gene factor 3(ISGF-3),and thereby reducing IAV-triggered pro-inflammatory response amplification in IFN-β-sensitized cells.Additionally,IAV-or vRNA-triggered apoptosis of alveolar epithelial A549 cells was prevented by erucic acid.In vivo,erucic acid administration consistently displayed decreased lung viral load and viral antigens expression.Meanwhile,erucic acid markedly reduced CD8+cytotoxic T lymphocyte(CTL)recruitment,pro-apoptotic signaling,hyperactivity of multiple signaling pathways,and exacerbated immune inflammation in the lung,which resulted in decreased lung injury and mortality in mice with a mouse-adapted A/FM/1/47-MA(H1N1)strain infection.Our findings provided a mechanistic basis for the action of erucic acid against IAV-mediated inflammation and injury,suggesting that erucic acid may have a therapeutic potential in the treatment of influenza.展开更多
Phenolic compounds, metabolites of the phenylpropanoid pathway, play an important role in the growth and environmental adaptation of many plants. Phenylalanine ammonia-lyase(PAL) is the first key enzyme of the phenylp...Phenolic compounds, metabolites of the phenylpropanoid pathway, play an important role in the growth and environmental adaptation of many plants. Phenylalanine ammonia-lyase(PAL) is the first key enzyme of the phenylpropanoid pathway. The present study was designed to investigate whether there is a multi-gene family in I. Indigotic and, if so, to characterize their properties. We conducted a comprehensive survey on the transcription profiling database by using tBLASTn analysis. Several bioinformatics methods were employed to perform the prediction of composition and physicochemical characters. The expression levels of IiPAL genes in various tissues of I. indigotica with stress treatment were examined by quantitative real-time PCR. Protoplast transient transformation was used to observe the locations of IiPALs. IiPALs were functionally characterized by expression with pET-32a vector in Escherichia colis strain BL21(DE3). Integration of transcripts and metabolite accumulations was used to reveal the relation between IiPALs and target compounds. An new gene(IiPAL2) was identified and both IiPALs had the conserved enzymatic active site Ala-Ser-Gly and were classified as members of dicotyledon. IiPAL1 and IiPAL2 were expressed in roots, stems, leaves, and flowers, with the highest expression levels of IiPAL1 and IiPAL2 being observed in stems and roots, respectively. The two genes responded to the exogenous elicitor in different manners. Subcellular localization experiment showed that both IiPALs were localized in the cytosol. The recombinant proteins were shown to catalyze the conversion of L-Phe to trans-cinnamic acid. Correlation analysis indicated that Ii PAL1 was more close to the biosynthesis of secondary metabolites than IiPAL2. In conclusion, the present study provides a basis for the elucidation of the role of Ii PALs genes in the biosynthesis of phenolic compounds, which will help further metabolic engineering to improve the accumulation of bioactive components in I.indigotica.展开更多
A glucosidic indole-lignan conjugate with a novel carbon skeleton, named isatindolignanoside A（1）, was isolated from an aqueous extract of the Isatis indigotica roots ＂ban lan gen＂. Its structure was elucidated by...A glucosidic indole-lignan conjugate with a novel carbon skeleton, named isatindolignanoside A（1）, was isolated from an aqueous extract of the Isatis indigotica roots ＂ban lan gen＂. Its structure was elucidated by comprehensive analysis of spectroscopic data, including J-and NOESY correlation-based configurational analysis, circular dichroism（CD） data, enzyme hydrolysis, and theoretical ECD calculation. In a preliminary assay, compound 1 showed antiviral activity against Coxsackie virus B3. This compound is the first example of natural product having a structural feature of conjugation between indole and lignan units, and its biosynthetic pathway is postulated.展开更多
Two pairs of unusual scalemic enantiomers with a dimethoxyphenyl）ethyl]-2-methoxyindolin-3-one, novel carbon skeleton of 2-[1′-（4＂-hydroxy-3＂,5＂- named isatidifoliumindolinones A-D （1-4）, were isolated from an...Two pairs of unusual scalemic enantiomers with a dimethoxyphenyl）ethyl]-2-methoxyindolin-3-one, novel carbon skeleton of 2-[1′-（4＂-hydroxy-3＂,5＂- named isatidifoliumindolinones A-D （1-4）, were isolated from an aqueous extract of lsatis indigotica leaves （da qing ye）. Their structures including absolute configurations were determined by spectroscopic data analysis combined with comparison of their experimental CD and calculated ECD spectra. Validity of the ECD spectra calculation to assign the absolute configurations is discussed. Plausible biosynthetic pathways of 1-4 are proposed. Stereochemistry-dependent activity against LPS-induced NO production in BY2 cells was observed, and among the stereoisomers compound 4 is most active.展开更多
Four stereoisomers of 3,5-bis（2-hydroxybut-3-en-1-yl）-l,2,4-thiadiazole, named insatindigothiadia- zoles A-D （1a-1d）, were isolated from the roots oflsatis indigotica. Their structures were determined by spectrosc...Four stereoisomers of 3,5-bis（2-hydroxybut-3-en-1-yl）-l,2,4-thiadiazole, named insatindigothiadia- zoles A-D （1a-1d）, were isolated from the roots oflsatis indigotica. Their structures were determined by spectroscopic analysis; specifically, the absolute configurations were assigned by using the MPA determination rule based on △δrs values of MPA esters, and supported by electronic CD （ECD） calculations. Proposed biosynthetic pathways and preliminary investigations of the biological activities of la-1d against influenza virus A （H3N2）, Coxsackie virus B3, and/or HSV-1 are also discussed.展开更多
A E-2-[(3'-indole)cyanomethylene]-3-indolinone was isolated from the root of Isatis indigotica Fort. The structure elucidation and 1H, 13C NMR assignments were achieved by spectroscopic method.
Objective AP2/ERF(APETALA2/ethylene-responsive factor)superfamily is one of the largest gene families in plants and has been reported to participate in various biological processes,such as the regulation of biosynthes...Objective AP2/ERF(APETALA2/ethylene-responsive factor)superfamily is one of the largest gene families in plants and has been reported to participate in various biological processes,such as the regulation of biosynthesis of active lignan.However,few studies have investigated the genome-wide role of the AP2/ERF superfamily in Isatis indigotica.This study establishes a complete picture of the AP2/ERF superfamily in I.indigotica and contributes valuable information for further functional characterization of IiAP2/ERF genes and supports further metabolic engineering.Methods To identify the IiAP2/ERF superfamily genes,the AP2/ERF sequences from Arabidopsis thaliana and Brassica rapa were used as query sequences in the basic local alignment search tool.Bioinformatic analyses were conducted to investigate the protein structure,motif composition,chromosome location,phylogenetic relationship,and interaction network of the IiAP2/ERF superfamily genes.The accuracy of omics data was verified by quantitative polymerase chain reaction and heatmap analyses.Results One hundred and twenty-six putative IiAP2/ERF genes in total were identified from the I.indigotica genome database in this study.By sequence alignment and phylogenetic analysis,the IiAP2/ERF genes were classified into 5 groups including AP2,ERF,DREB(dehydration-responsive element-binding factor),Soloist and RAV(related to abscisic acid insensitive 3/viviparous 1)subfamilies.Among which,122 members were unevenly distributed across seven chromosomes.Sequence alignment showed that I.indigotica and A.thaliana had 30 pairs of orthologous genes,and we constructed their interaction network.The comprehensive analysis of gene expression pattern in different tissues suggested that these genes may play a significant role in organ growth and development of I.indigotica.Members that may regulate lignan biosynthesis in roots were also preliminarily identified.Ribonucleic acid sequencing analysis revealed that the expression of 76 IiAP2/ERF genes were up-or down-regulated under salt or drought treatment,among which,33 IiAP2/ERF genes were regulated by both stresses.Conclusion This study undertook a genome-wide characterization of the AP2/ERF superfamily in I.indigotica,providing valuable information for further functional characterization of IiAP2/ERF genes and discovery of genetic targets for metabolic engineering.展开更多
Climate change is affecting global crop productivity, food quality, and security. However,few studies have addressed the mechanism by which elevated CO_2 may affect the growth of medicinal plants. Isatis indigotica Fo...Climate change is affecting global crop productivity, food quality, and security. However,few studies have addressed the mechanism by which elevated CO_2 may affect the growth of medicinal plants. Isatis indigotica Fortune is a widely used Chinese medicinal herb with multiple pharmacological properties. To investigate the physiological mechanism of I.indigotica response to elevated [CO_2], plants were grown at either ambient [CO_2](385 μmol mol^(-1)) or elevated [CO_2] (590 μmol mol^(-1)) in an open-top chamber (OTC)experimental facility in North China. A significant reduction in transpiration rate (T_r) and stomatal conductance (g_s) and a large increase in water-use efficiency contributed to an increase in net photosynthetic rate (Pn) under elevated [CO_2] 76 days after sowing. Leaf non-photochemical quenching (NPQ) was decreased, so that more energy was used in effective quantum yield of PSII photochemistry (Φ_(PSⅡ)) under elevated [CO_2]. High ΦPSII,meaning high electron transfer efficiency, also increased Pn. The [CO_2]-induced increase in photosynthesis significantly increased biomass by 36.8%. Amounts of metabolic compounds involved in sucrose metabolism, pyrimidine metabolism, flavonoid biosynthesis, and other processes in leaves were reduced under elevated [CO_2]. These results showed that the fertilization effect of elevated [CO_2] is conducive to increasing dry weight but not secondary metabolism in I. indigotica.展开更多
A pair of indole alkaloid enantiomers with a novel bisindolylacetamide skeleton, insatindibisindolamides A and B（1a and 1b）, was isolated from an aqueous extract of Isatis indigotica roots. The enantiomers were sepa...A pair of indole alkaloid enantiomers with a novel bisindolylacetamide skeleton, insatindibisindolamides A and B（1a and 1b）, was isolated from an aqueous extract of Isatis indigotica roots. The enantiomers were separated by chiral HPLC. Their structures and absolute configurations were elucidated by extensive spectroscopic analysis, including 2D NMR, X-ray crystallography, and electronic CD（ECD） calculation. The proposed biosynthetic pathway and preliminary investigations of the biological activity of compounds 1a and 1b are also discussed.展开更多
This study is aimed to validate the extraction process of Isatis Radix. Using near infrared spectroscopy and high performance liquid chromatography to determine the uridine, adenosine, guanosine contents in Isatis Rad...This study is aimed to validate the extraction process of Isatis Radix. Using near infrared spectroscopy and high performance liquid chromatography to determine the uridine, adenosine, guanosine contents in Isatis Radix Solution, as well as NIR to monitor the extraction process. The results indicate that the existing extraction is optimized to extract thoroughly.展开更多
Isaindigotone 1 and 3-(2'-hydroxyphenyl)-4(3H)-quinazolinone 2 were isolated from the roots of Isatis indigotica. Compound 1 was elucidated as a new alkaloid.Compound 2 was discovered for the first time from a nat...Isaindigotone 1 and 3-(2'-hydroxyphenyl)-4(3H)-quinazolinone 2 were isolated from the roots of Isatis indigotica. Compound 1 was elucidated as a new alkaloid.Compound 2 was discovered for the first time from a natural source展开更多
Objective： To study and analyze the antibacterial effects of different extracts from Radix Isatis. Methods： Staphylococcus aureus was used as the studied object in the experiment. Antibacterial effects of extracts f...Objective： To study and analyze the antibacterial effects of different extracts from Radix Isatis. Methods： Staphylococcus aureus was used as the studied object in the experiment. Antibacterial effects of extracts from Radix Isatis were observed by thermocalrimetry on Staphylococcus aureus, together with common pharmacological experiments. Results： The total extract, ethyl acetate （EtOAc） extract, n-butylalcohol （nBuOH） extract, chloroform （CHCl3） extract and petroleum （P. E.） extract had antiviral effects to some extent while the residue after extracting had no antibacterial activity. The potency of antiviral activity among them was as follows： nBuOH extract 〉 EtOAc extract 〉 CHCl3 extract〉 total extract 〉P. E. extract. Conclusion： The antibacteriall effects of Radix Isatis were not limited to any active portion, showing that Radix Isatis exerts its antibacterial effects by cooperation of different active fractions in varied ways.展开更多
Seven alkaloids including five undescribed ones(1 a/1 b,2,3 and 5)were obtained from the leaves of Isatis indigotica Fortune.Their structures were established by extensive spectroscopic analyses.The absolute configura...Seven alkaloids including five undescribed ones(1 a/1 b,2,3 and 5)were obtained from the leaves of Isatis indigotica Fortune.Their structures were established by extensive spectroscopic analyses.The absolute configurations of compounds 1 a,1 b,3 and 5 were determined by comparison of the experimental and calculated electronic circular dichroism(ECD)spectra.Subsequently,the neuroprotective effects of all the isolates against H_(2)O_(2)-induced injury in SH-SY5 Y cells were evaluated in vitro by MTT assay.Moreover,Annexin V-FITC/PI double staining was performed,while the activities of antioxidant enzymes(SOD,CAT and GSH-Px)for compounds 1 a and 1 b were measured.展开更多
Objectives: Isatis tinctoria L., clary, is an herbal plant traditionally used in folk medicine for the treatment of various diseases and conditions. Although it has been primarily used as an antimicrobial and antifung...Objectives: Isatis tinctoria L., clary, is an herbal plant traditionally used in folk medicine for the treatment of various diseases and conditions. Although it has been primarily used as an antimicrobial and antifungal, there are data on traditional use of I. tinctoria as an agent against antiallergic, anti-thrombocytosis. The aim of the study was to examine the effect of the I. tinctoria root extract on the lipopolysaccharide (LPS)-induced periodontitis in rats on osteoclast associated bone resorptive activity, cell death including apoptosis, and inflammation in a rat of disease model of periodontitis. Materials and Methods: Periodontitis, acute or chronic inflammatory status in periodontal tissue in rats could be induced by repeated injections of LPS from Escherichia coli into the periodontal pocket area between the first and second right maxillary molars. Eighteen male rats were distributed among the following treatment groups: 1) I. tinctorial root extract (Antifect) 200 mg/kg body weight, 2) acetylsalicylic acid (ASA), 20 mg/kg body weight and 3) Phosphate buffered saline (PBS) treatment used as a control. After 15 days, maxilla, alveolar bone, molar teeth and associated periodontal tissues were harvested. Inflammatory alveolar bone resorption was analyzed by microcomputerized tomography (μCT) (microcomputer tomography). Tissues fixed with paraformaldehyde and formalin for 2 days, after that paraffin embedded histological sections were stained with haematoxylin and eosin (H/E) for the assessment of histopathological changes or tested to immunohistochemistry for detecting TRAP (tartrate resistant acid phosphatase) positive cells and caspase 3. Cell death and Apoptosis were analyzed in the periodontal tissues by tunnel assay. The inflammatory status was assessed by the measurements of proinflammatory cytokines interleukin-Iβ (IL-Iβ), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) of gingival tissues and descriptive analysis of histological sections of periodontal. Results: Treatment with Antifect, compared to the control group, significantly diminished the process of inflammation decreasing the levels of IL-Iβ, IL-6 and TNF-α, reducing the gingival tissue lesions and preserving bone alveolar resorption. Considerably a smaller number of inflammatory cells and a larger number of fibroblasts were noticed. Also, μCT analysis showed that only Antifect treated group reduced bone resorption and the number of TRAP-positive multinucleated cells (osteoclasts), also, significantly reduced the number of apoptotic cells in the gingival tissues and of osteocytes in the alveolar bone crest. Conclusion: Antifect manifested anti-inflammatory elect and reducing alveolar bone resorption in LPS-induced periodontitis suggest that it may have a role as a therapeutic agent in periodontal diseases.展开更多
A phytochemical investigation of 80%EtOH exact of Isatis indigotica Fortune leaves yielded ten lignans(1-10),including six furofurans lignans(1-6),one arylnaphthalenes lignans(7)and three tetrahydrofurans lignans(8-10...A phytochemical investigation of 80%EtOH exact of Isatis indigotica Fortune leaves yielded ten lignans(1-10),including six furofurans lignans(1-6),one arylnaphthalenes lignans(7)and three tetrahydrofurans lignans(8-10).Their structures were determined by HRESIMS spectroscopic data and extensive NMR analyses.Among them,compounds 6,9 and 10 were isolated from the genus oi Isatis for the first time.展开更多
Phenylpropanoid derivatives are a complex class of secondary metabolites that have many important roles in plants during normal growth and in responses to environmental stress. Phenylalanine ammonialyase （PAL） catal...Phenylpropanoid derivatives are a complex class of secondary metabolites that have many important roles in plants during normal growth and in responses to environmental stress. Phenylalanine ammonialyase （PAL） catalyzes the first step in the biosynthesis of phenylpropanoids. In the present study, we isolated a novel phenylalanine ammonialyase gene （designated as liPAL） from tetraploid Isatis indigotica Fort. by rapid amplification of cDNA ends （RACE）, which was a cultivar from the diploid plant by genome duplication. The full-length cDNA of liPAL was 2 530-bp long with an open reading frame （ORF） of 2 178 bp encoding a polypeptide of 725 amino acid residues. Analysis of liPAL genomic DNA revealed that it was structurally similar to other plant PAL genes, with a single intron at a conserved position, and a long highly conserved second exon. Semi-quantitative RT-PCR revealed that the liPAL expression in roots and leaves from a tetraploid sample was higher than that in diploid progenitor, whereas expression of liPAL in stems was almost the same as each other. Furthermore, the highest expression of liPAL in tetraploid plant was found in roots, which was found in stems in diploid plants. Further expression analysis revealed that gibberelUn （GA3）, abscisic acid （ABA）, methyl jasmonate （MeJA） and cold treatments could up-regulate the liPAL transcription in tetraploid plants. All our findings suggest that liPAL participates not only in the defense/stress responsive pathways, but also probably in the polyploidy evolution of L indigotica.展开更多
Abstract: The seed embryos of Isatis indigotica Fort were exposed to He-Ne laser (5.23 mW/mm2, radiated for 5 min) and microwave (1.26 mW/mm2, radiated for 8 s) irradiation to determine the effects of microwave and He...Abstract: The seed embryos of Isatis indigotica Fort were exposed to He-Ne laser (5.23 mW/mm2, radiated for 5 min) and microwave (1.26 mW/mm2, radiated for 8 s) irradiation to determine the effects of microwave and He-Ne laser pretreatment on enzyme activities, and biophoton emission of cotyledon. Then: (i) changes in the activities of enzymes in I. indigotica cotyledon (such as amylase, transaminase, and proteinase) were measured to investigate the effects of He-Ne laser and microwave pretreatment; and (ii) biophoton emission was measured to determine the speed of cell division and metabolism. Results from these experiments indicated that: (i) the activities of amylase, transaminase, and proteinase of the cotyledon pretreated by He-Ne laser and microwave were significantly increased; and (ii) the intensity of biophoton emission was enhanced significantly by He-Ne laser and microwave irradiation. These changes suggest that He-Ne laser and microwave pretreatment can improve the inner energy of seeds, lead to an enhancement of cotyledon enzymes, and speed up the metabolism of the cell, resulting in significantly increased biophoton emission. Moreover, the mechanism of action of the effects of laser and microwave radiation on the microcalorimetric parameters, enzyme activities, and biophoton emission of seeds is discussed on the basis of the results obtained.展开更多
文摘This article is a summary of the historical background to Isatis tinctoria in France and elsewhere from the 12th to the 21st century. Isatis tinctoria, also called “Guède”, “Pastel” or “Vouède” according to the French region origin, is a plant originally used to obtain indigo blue dye mainly to produce colored blue fabrics. A local memory is well established in the Amiens region (northern France) and the Toulouse region (south-western France). No global review of its culture and trade over the long term is known. The aim of this article is to summarize the historical background of Isatis tinctoria in France and elsewhere from the twelfth to the twenty-first century.
文摘[Objective] This study aimed to compare the microstructure between I. in- digotica Fort. and B. cusia （Nees） Bremek. [Method] Microstructure of I. indigotica Fort. and B. cusia （Nees） Bremek. with different sources were compared and analyzed by using digital microscopy. [Result] Comparison result of the microstructure shows that reticulate vessels, single starch granules, multiple starch granules and stone cells are mostly observed in I. indigotica Fort.; pitted vessels, parenchyma cells with small pits, single starch granules and cystolith are mostly observed in B. cusia （Nees） Bremek., which indicated that microstructure of L indigotica Fort. and B. cusia （Nees） Bremek. are significantly different. [Conclusion] Microstructure char- acteristics of I. indigotica Fort. and B. cusia （Nees） Bremek. established in this study can be used to evaluate the quality of these two kinds of Chinese herbal medicines, which also provide scientific basis and methods for comprehensively studying the quality standard of /. indigotica Fort. and B. cusia （Nees） Bremek. Key words Isatis indigotica Fort.; Baphicacanthus cusia （Nees） Bremek.; Microstruc- ture; Quality
基金the Doctoral Research Initiation Foundation of Changzhi Medical College(BS202005)。
文摘This study was to investigate the effects of three exogenous substances on chemical constituents of Isatis indigotica leavesand their efficacy in alleviating drought stress, and explore the methods of applying exogenous substances to efficient cultivationof Isatis indigotica. Polyethylene glycol (PEG) was used to simulate drought stress to deal with seeds of Isatis indigotica at thegermination stage (concentration: 0, 10%, 15%, and 20%). Simultaneous operation of exogenous growth regulators [microbialinoculum (MI), γ-aminobutyric acid (GABA) and salicylic acid (SA)] and PEG were implemented in seeds of Isatis indigotica.The effects of drought stress and the mitigation of exogenous substances were observed by statistics of seed germination potential,germination rate, hypocotyl length, and radicle length of each treatment. The effects of exogenous substances on the content ofalkaloids, crude protein and free amino acids in the leaves of Isatis indigotica grown in a greenhouse were determined after sprayingexogenous substances on the plants. The differences of germination potential, germination rate, hypocotyl length, and radicle lengthamong 15% PEG stress treatment, 10% PEG stress treatment and the control were significant (P<0.05). According to the predesignedgermination standard, the seeds did not germinate under 20% PEG stress treatment. When the PEG concentration was 15%, the resultsof seed germination potential and germination rate after adding MI were significantly different from those under stress alone (P<0.05).When exposed to 10% PEG stress, the supplementation of GABA led to a notable increase in radicle length of Isatis indigotica seeds,showing significant differences compared to other three treatments. The application of MI and GABA under 15% PEG stress resultedin a significant increase in the radicle and hypocotyl length of Isatis indigotica seeds compared to other two treatments. The contentof the total alkaloids in leaves of Isatis indigotica was significantly increased after spraying GABA. Meanwhile, the contents of crudeprotein and the total free amino acids were kept constant after spraying exogenous substances. Application of MI and GABA couldalleviate drought stress of Isatis indigotica. The content of the total alkaloids in leaves of Isatis indigotica could significantly increaseafter spraying GABA.
基金funded by the National Natural Science Foundation of China(Grantno.81873065)the Secondary Development Projects of Guangdong Famous and Excellent TraditionalChinese Patent Medicines(Grant no.20174005)+1 种基金the Natural Science Foundation of Guangdong Province(Grant no.2018A030310172)the China Postdoctoral Science Foundation(Grant no.2017M622652,2019M652987)。
文摘Isatis indigotica Fort.(Ban-Lan-Gen)is an herbal medicine prescribed for influenza treatment.However,its active components and mode of action remain mostly unknown.In the present study,erucic acid was isolated from Isatis indigotica Fort.,and subsequently its underlying mechanism against influenza A virus(IAV)infection was investigated in vitro and in vivo.Our results demonstrated that erucic acid exhibited broad-spectrum antiviral activity against IAV resulting from reduction of viral polymerase transcription activity.Erucic acid was found to exert inhibitory effects on IAV or viral(v)RNA-induced pro-inflam-matory mediators as well as interferons(IFNs).The molecular mechanism by which erucic acid with antiviral and anti-inflammatory properties was attributed to inactivation of NF-kB and p38 MAPK signaling.Furthermore,the NF-kB and p38 MAPK inhibitory effect of erucic acid led to diminishing the transcriptional activity of interferon-stimulated gene factor 3(ISGF-3),and thereby reducing IAV-triggered pro-inflammatory response amplification in IFN-β-sensitized cells.Additionally,IAV-or vRNA-triggered apoptosis of alveolar epithelial A549 cells was prevented by erucic acid.In vivo,erucic acid administration consistently displayed decreased lung viral load and viral antigens expression.Meanwhile,erucic acid markedly reduced CD8+cytotoxic T lymphocyte(CTL)recruitment,pro-apoptotic signaling,hyperactivity of multiple signaling pathways,and exacerbated immune inflammation in the lung,which resulted in decreased lung injury and mortality in mice with a mouse-adapted A/FM/1/47-MA(H1N1)strain infection.Our findings provided a mechanistic basis for the action of erucic acid against IAV-mediated inflammation and injury,suggesting that erucic acid may have a therapeutic potential in the treatment of influenza.
基金supported by the Natural Science Foundation of China(Nos.31100221 and 81325024)
文摘Phenolic compounds, metabolites of the phenylpropanoid pathway, play an important role in the growth and environmental adaptation of many plants. Phenylalanine ammonia-lyase(PAL) is the first key enzyme of the phenylpropanoid pathway. The present study was designed to investigate whether there is a multi-gene family in I. Indigotic and, if so, to characterize their properties. We conducted a comprehensive survey on the transcription profiling database by using tBLASTn analysis. Several bioinformatics methods were employed to perform the prediction of composition and physicochemical characters. The expression levels of IiPAL genes in various tissues of I. indigotica with stress treatment were examined by quantitative real-time PCR. Protoplast transient transformation was used to observe the locations of IiPALs. IiPALs were functionally characterized by expression with pET-32a vector in Escherichia colis strain BL21(DE3). Integration of transcripts and metabolite accumulations was used to reveal the relation between IiPALs and target compounds. An new gene(IiPAL2) was identified and both IiPALs had the conserved enzymatic active site Ala-Ser-Gly and were classified as members of dicotyledon. IiPAL1 and IiPAL2 were expressed in roots, stems, leaves, and flowers, with the highest expression levels of IiPAL1 and IiPAL2 being observed in stems and roots, respectively. The two genes responded to the exogenous elicitor in different manners. Subcellular localization experiment showed that both IiPALs were localized in the cytosol. The recombinant proteins were shown to catalyze the conversion of L-Phe to trans-cinnamic acid. Correlation analysis indicated that Ii PAL1 was more close to the biosynthesis of secondary metabolites than IiPAL2. In conclusion, the present study provides a basis for the elucidation of the role of Ii PALs genes in the biosynthesis of phenolic compounds, which will help further metabolic engineering to improve the accumulation of bioactive components in I.indigotica.
基金Financial support from the National Natural Science Foundation of China (Nos. 81373287, 81630094, and 30825044) is acknowledged
文摘A glucosidic indole-lignan conjugate with a novel carbon skeleton, named isatindolignanoside A（1）, was isolated from an aqueous extract of the Isatis indigotica roots ＂ban lan gen＂. Its structure was elucidated by comprehensive analysis of spectroscopic data, including J-and NOESY correlation-based configurational analysis, circular dichroism（CD） data, enzyme hydrolysis, and theoretical ECD calculation. In a preliminary assay, compound 1 showed antiviral activity against Coxsackie virus B3. This compound is the first example of natural product having a structural feature of conjugation between indole and lignan units, and its biosynthetic pathway is postulated.
基金Financial support from the National Natural Science Foundation of China (NNSFCNos.81630094,81373287,and 30825044)
文摘Two pairs of unusual scalemic enantiomers with a dimethoxyphenyl）ethyl]-2-methoxyindolin-3-one, novel carbon skeleton of 2-[1′-（4＂-hydroxy-3＂,5＂- named isatidifoliumindolinones A-D （1-4）, were isolated from an aqueous extract of lsatis indigotica leaves （da qing ye）. Their structures including absolute configurations were determined by spectroscopic data analysis combined with comparison of their experimental CD and calculated ECD spectra. Validity of the ECD spectra calculation to assign the absolute configurations is discussed. Plausible biosynthetic pathways of 1-4 are proposed. Stereochemistry-dependent activity against LPS-induced NO production in BY2 cells was observed, and among the stereoisomers compound 4 is most active.
基金Financial support from the National Natural Science Foundation of China (Nos. 81373287 and 30825044)the Beijing Excellent Talent Training Project (No. 2013D009008000002)the National Science and Technology Project of China (Nos. 2012ZX09301002002 and 2011ZX09307-002-01)
文摘Four stereoisomers of 3,5-bis（2-hydroxybut-3-en-1-yl）-l,2,4-thiadiazole, named insatindigothiadia- zoles A-D （1a-1d）, were isolated from the roots oflsatis indigotica. Their structures were determined by spectroscopic analysis; specifically, the absolute configurations were assigned by using the MPA determination rule based on △δrs values of MPA esters, and supported by electronic CD （ECD） calculations. Proposed biosynthetic pathways and preliminary investigations of the biological activities of la-1d against influenza virus A （H3N2）, Coxsackie virus B3, and/or HSV-1 are also discussed.
基金the National Natural Science Foundation of China (39670878)
文摘A E-2-[(3'-indole)cyanomethylene]-3-indolinone was isolated from the root of Isatis indigotica Fort. The structure elucidation and 1H, 13C NMR assignments were achieved by spectroscopic method.
基金This study was supported by the National Natural Science Foundation of China(No.32000231,31970316,32170274,31770329 and 32070327)Program of Shanghai Academic Research Leader(19XD1405000)Shanghai Sail Program(19YF1459300)。
文摘Objective AP2/ERF(APETALA2/ethylene-responsive factor)superfamily is one of the largest gene families in plants and has been reported to participate in various biological processes,such as the regulation of biosynthesis of active lignan.However,few studies have investigated the genome-wide role of the AP2/ERF superfamily in Isatis indigotica.This study establishes a complete picture of the AP2/ERF superfamily in I.indigotica and contributes valuable information for further functional characterization of IiAP2/ERF genes and supports further metabolic engineering.Methods To identify the IiAP2/ERF superfamily genes,the AP2/ERF sequences from Arabidopsis thaliana and Brassica rapa were used as query sequences in the basic local alignment search tool.Bioinformatic analyses were conducted to investigate the protein structure,motif composition,chromosome location,phylogenetic relationship,and interaction network of the IiAP2/ERF superfamily genes.The accuracy of omics data was verified by quantitative polymerase chain reaction and heatmap analyses.Results One hundred and twenty-six putative IiAP2/ERF genes in total were identified from the I.indigotica genome database in this study.By sequence alignment and phylogenetic analysis,the IiAP2/ERF genes were classified into 5 groups including AP2,ERF,DREB(dehydration-responsive element-binding factor),Soloist and RAV(related to abscisic acid insensitive 3/viviparous 1)subfamilies.Among which,122 members were unevenly distributed across seven chromosomes.Sequence alignment showed that I.indigotica and A.thaliana had 30 pairs of orthologous genes,and we constructed their interaction network.The comprehensive analysis of gene expression pattern in different tissues suggested that these genes may play a significant role in organ growth and development of I.indigotica.Members that may regulate lignan biosynthesis in roots were also preliminarily identified.Ribonucleic acid sequencing analysis revealed that the expression of 76 IiAP2/ERF genes were up-or down-regulated under salt or drought treatment,among which,33 IiAP2/ERF genes were regulated by both stresses.Conclusion This study undertook a genome-wide characterization of the AP2/ERF superfamily in I.indigotica,providing valuable information for further functional characterization of IiAP2/ERF genes and discovery of genetic targets for metabolic engineering.
基金partially supported by the National Natural Science Foundation of China (Nos. 31601212, 31371693, and 31471556)Research on Science and Technology of Shanxi Province (No. 20150311006-2)+1 种基金Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi (No. 2015146)the Shanxi 100-Talent Program
文摘Climate change is affecting global crop productivity, food quality, and security. However,few studies have addressed the mechanism by which elevated CO_2 may affect the growth of medicinal plants. Isatis indigotica Fortune is a widely used Chinese medicinal herb with multiple pharmacological properties. To investigate the physiological mechanism of I.indigotica response to elevated [CO_2], plants were grown at either ambient [CO_2](385 μmol mol^(-1)) or elevated [CO_2] (590 μmol mol^(-1)) in an open-top chamber (OTC)experimental facility in North China. A significant reduction in transpiration rate (T_r) and stomatal conductance (g_s) and a large increase in water-use efficiency contributed to an increase in net photosynthetic rate (Pn) under elevated [CO_2] 76 days after sowing. Leaf non-photochemical quenching (NPQ) was decreased, so that more energy was used in effective quantum yield of PSII photochemistry (Φ_(PSⅡ)) under elevated [CO_2]. High ΦPSII,meaning high electron transfer efficiency, also increased Pn. The [CO_2]-induced increase in photosynthesis significantly increased biomass by 36.8%. Amounts of metabolic compounds involved in sucrose metabolism, pyrimidine metabolism, flavonoid biosynthesis, and other processes in leaves were reduced under elevated [CO_2]. These results showed that the fertilization effect of elevated [CO_2] is conducive to increasing dry weight but not secondary metabolism in I. indigotica.
基金Financial support from the National Natural Science Foundation of China(NNSFCNos.81373287 and 30825044)+1 种基金the Program for Changjiang Scholars and Innovative Research Team in University(PCSIRT,No.IRT1007)the National Science and Technology Project of China(Nos.2012ZX09301002-002 and 2011ZX0 9307-002-01)
文摘A pair of indole alkaloid enantiomers with a novel bisindolylacetamide skeleton, insatindibisindolamides A and B（1a and 1b）, was isolated from an aqueous extract of Isatis indigotica roots. The enantiomers were separated by chiral HPLC. Their structures and absolute configurations were elucidated by extensive spectroscopic analysis, including 2D NMR, X-ray crystallography, and electronic CD（ECD） calculation. The proposed biosynthetic pathway and preliminary investigations of the biological activity of compounds 1a and 1b are also discussed.
文摘This study is aimed to validate the extraction process of Isatis Radix. Using near infrared spectroscopy and high performance liquid chromatography to determine the uridine, adenosine, guanosine contents in Isatis Radix Solution, as well as NIR to monitor the extraction process. The results indicate that the existing extraction is optimized to extract thoroughly.
文摘Isaindigotone 1 and 3-(2'-hydroxyphenyl)-4(3H)-quinazolinone 2 were isolated from the roots of Isatis indigotica. Compound 1 was elucidated as a new alkaloid.Compound 2 was discovered for the first time from a natural source
基金Supported by Foundation of State Administration of Tradi-tional Chinese Medicine (No .04-05ZP70)
文摘Objective： To study and analyze the antibacterial effects of different extracts from Radix Isatis. Methods： Staphylococcus aureus was used as the studied object in the experiment. Antibacterial effects of extracts from Radix Isatis were observed by thermocalrimetry on Staphylococcus aureus, together with common pharmacological experiments. Results： The total extract, ethyl acetate （EtOAc） extract, n-butylalcohol （nBuOH） extract, chloroform （CHCl3） extract and petroleum （P. E.） extract had antiviral effects to some extent while the residue after extracting had no antibacterial activity. The potency of antiviral activity among them was as follows： nBuOH extract 〉 EtOAc extract 〉 CHCl3 extract〉 total extract 〉P. E. extract. Conclusion： The antibacteriall effects of Radix Isatis were not limited to any active portion, showing that Radix Isatis exerts its antibacterial effects by cooperation of different active fractions in varied ways.
基金supported by the National Natural Science Foundation of China(Nos.82003635 and 81973462)the Liaoning Revitalization Talents Program(No.XLYC2007180)+2 种基金the Program for Major Scientific and Medical Technology Problems of China Medicine Education Association in 2020(No.2020KTS003)the Natural Science Foundation of Liaoning Province of China(No.2019-MS-303)the Scientific Research Fund Project of Liaoning Provincial Department of Education(No.2019LJC05)。
文摘Seven alkaloids including five undescribed ones(1 a/1 b,2,3 and 5)were obtained from the leaves of Isatis indigotica Fortune.Their structures were established by extensive spectroscopic analyses.The absolute configurations of compounds 1 a,1 b,3 and 5 were determined by comparison of the experimental and calculated electronic circular dichroism(ECD)spectra.Subsequently,the neuroprotective effects of all the isolates against H_(2)O_(2)-induced injury in SH-SY5 Y cells were evaluated in vitro by MTT assay.Moreover,Annexin V-FITC/PI double staining was performed,while the activities of antioxidant enzymes(SOD,CAT and GSH-Px)for compounds 1 a and 1 b were measured.
文摘Objectives: Isatis tinctoria L., clary, is an herbal plant traditionally used in folk medicine for the treatment of various diseases and conditions. Although it has been primarily used as an antimicrobial and antifungal, there are data on traditional use of I. tinctoria as an agent against antiallergic, anti-thrombocytosis. The aim of the study was to examine the effect of the I. tinctoria root extract on the lipopolysaccharide (LPS)-induced periodontitis in rats on osteoclast associated bone resorptive activity, cell death including apoptosis, and inflammation in a rat of disease model of periodontitis. Materials and Methods: Periodontitis, acute or chronic inflammatory status in periodontal tissue in rats could be induced by repeated injections of LPS from Escherichia coli into the periodontal pocket area between the first and second right maxillary molars. Eighteen male rats were distributed among the following treatment groups: 1) I. tinctorial root extract (Antifect) 200 mg/kg body weight, 2) acetylsalicylic acid (ASA), 20 mg/kg body weight and 3) Phosphate buffered saline (PBS) treatment used as a control. After 15 days, maxilla, alveolar bone, molar teeth and associated periodontal tissues were harvested. Inflammatory alveolar bone resorption was analyzed by microcomputerized tomography (μCT) (microcomputer tomography). Tissues fixed with paraformaldehyde and formalin for 2 days, after that paraffin embedded histological sections were stained with haematoxylin and eosin (H/E) for the assessment of histopathological changes or tested to immunohistochemistry for detecting TRAP (tartrate resistant acid phosphatase) positive cells and caspase 3. Cell death and Apoptosis were analyzed in the periodontal tissues by tunnel assay. The inflammatory status was assessed by the measurements of proinflammatory cytokines interleukin-Iβ (IL-Iβ), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) of gingival tissues and descriptive analysis of histological sections of periodontal. Results: Treatment with Antifect, compared to the control group, significantly diminished the process of inflammation decreasing the levels of IL-Iβ, IL-6 and TNF-α, reducing the gingival tissue lesions and preserving bone alveolar resorption. Considerably a smaller number of inflammatory cells and a larger number of fibroblasts were noticed. Also, μCT analysis showed that only Antifect treated group reduced bone resorption and the number of TRAP-positive multinucleated cells (osteoclasts), also, significantly reduced the number of apoptotic cells in the gingival tissues and of osteocytes in the alveolar bone crest. Conclusion: Antifect manifested anti-inflammatory elect and reducing alveolar bone resorption in LPS-induced periodontitis suggest that it may have a role as a therapeutic agent in periodontal diseases.
文摘A phytochemical investigation of 80%EtOH exact of Isatis indigotica Fortune leaves yielded ten lignans(1-10),including six furofurans lignans(1-6),one arylnaphthalenes lignans(7)and three tetrahydrofurans lignans(8-10).Their structures were determined by HRESIMS spectroscopic data and extensive NMR analyses.Among them,compounds 6,9 and 10 were isolated from the genus oi Isatis for the first time.
基金Supported by the National Natural Science Foundation of China （30371746）,
文摘Phenylpropanoid derivatives are a complex class of secondary metabolites that have many important roles in plants during normal growth and in responses to environmental stress. Phenylalanine ammonialyase （PAL） catalyzes the first step in the biosynthesis of phenylpropanoids. In the present study, we isolated a novel phenylalanine ammonialyase gene （designated as liPAL） from tetraploid Isatis indigotica Fort. by rapid amplification of cDNA ends （RACE）, which was a cultivar from the diploid plant by genome duplication. The full-length cDNA of liPAL was 2 530-bp long with an open reading frame （ORF） of 2 178 bp encoding a polypeptide of 725 amino acid residues. Analysis of liPAL genomic DNA revealed that it was structurally similar to other plant PAL genes, with a single intron at a conserved position, and a long highly conserved second exon. Semi-quantitative RT-PCR revealed that the liPAL expression in roots and leaves from a tetraploid sample was higher than that in diploid progenitor, whereas expression of liPAL in stems was almost the same as each other. Furthermore, the highest expression of liPAL in tetraploid plant was found in roots, which was found in stems in diploid plants. Further expression analysis revealed that gibberelUn （GA3）, abscisic acid （ABA）, methyl jasmonate （MeJA） and cold treatments could up-regulate the liPAL transcription in tetraploid plants. All our findings suggest that liPAL participates not only in the defense/stress responsive pathways, but also probably in the polyploidy evolution of L indigotica.
文摘Abstract: The seed embryos of Isatis indigotica Fort were exposed to He-Ne laser (5.23 mW/mm2, radiated for 5 min) and microwave (1.26 mW/mm2, radiated for 8 s) irradiation to determine the effects of microwave and He-Ne laser pretreatment on enzyme activities, and biophoton emission of cotyledon. Then: (i) changes in the activities of enzymes in I. indigotica cotyledon (such as amylase, transaminase, and proteinase) were measured to investigate the effects of He-Ne laser and microwave pretreatment; and (ii) biophoton emission was measured to determine the speed of cell division and metabolism. Results from these experiments indicated that: (i) the activities of amylase, transaminase, and proteinase of the cotyledon pretreated by He-Ne laser and microwave were significantly increased; and (ii) the intensity of biophoton emission was enhanced significantly by He-Ne laser and microwave irradiation. These changes suggest that He-Ne laser and microwave pretreatment can improve the inner energy of seeds, lead to an enhancement of cotyledon enzymes, and speed up the metabolism of the cell, resulting in significantly increased biophoton emission. Moreover, the mechanism of action of the effects of laser and microwave radiation on the microcalorimetric parameters, enzyme activities, and biophoton emission of seeds is discussed on the basis of the results obtained.
