Isatis Indigotica Research Articles

Artificial neural network techniques to predict the moisture ratio content during hot air drying and vacuum drying of Radix isatidis extract

Artificial neural network techniques to predict the moisture ratio content during hot air drying and vacuum drying of Radix isatidis extract

A valid evaluation method for UPLC fingerprint analysis and moisture ratio prediction model: application to microwave vacuum drying of Radix isatidis extract

A valid evaluation method for UPLC fingerprint analysis and moisture ratio prediction model: application to microwave vacuum drying of Radix isatidis extract

Polysaccharide Nanoparticles from Isatis indigotica Fort. Root Decoction: Diversity, Cytotoxicity, and Antiviral Activity.

It has been revealed that numerous nanoparticles are formed during the boiling preparation of traditional Chinese medical decoctions and culinary soups. They may possess physiological effects different from those of constituent components and are worth paying attention to but are barely noticed and investigated as of yet. In this study, six groups of nanoparticles, whose size ranged from 57 to 300 nm, were successfully isolated from the decoction of Isatis indigotica Fort. root, according to their particle size by the means of size-exclusive chromatography. All of the obtained nanoparticles have a high content of polysaccharides, which distinguishes them from the disclosed BLG protein nanoparticles. They also have high similarities in other compositions, surface charge, and stimuli responses. However, four out of these six nanoparticles (F2, F3, F4, and F5) exhibited significant antiviral activity against influenza virus H1N1, and their antiviral activities and cytotoxicity towards MDCK cells varied with their sizes. It suggested that the antiviral efficacy of BLG decoction could also be from its nanoparticles besides its well-known antiviral phytochemicals. It also implied that the biological effects of these polysaccharide nanoparticles, including cytotoxicity and antiviral activity, may be correlative with the physicochemical properties, especially the particle size.

The Effects of Fig Tree (Ficus carica L.) Leaf Aqueous Extract on Seed Germination and Seedling Growth of Three Medicinal Plants

Fig tree cultivation land resources are not fully utilized and introducing them into sustainable medicinal agroforestry systems can effectively achieve resource protection and reuse. Laboratory and pot experiments were applied to study the allelopathic effects of fig tree (Ficus carica L.) leaf aqueous extract at five mass concentrations of 8.3, 10.0, 12.5, 16.7, and 25.0 g/L on the morphological and physiological indexes of mint (Mentha haplocalyx Briq.), dandelion (Taraxacum mongolicum Hand.-Mazz.), and woad (Isatis indigotica Fort.). The results showed that mint had the best seed germination rate. The leaf aqueous extract at lower concentrations had a strong promoting effect on the biomass and photosynthetic parameters of mint, dandelion, and woad. With the increase in leaf aqueous extract concentration, the superoxide dismutase, peroxidase, and catalase activity of mint, dandelion, and woad increased initially and then decreased, but the malondialdehyde content increased. The synthetic allelopathic indexes of the three medicinal plants were in the following order: mint > woad > dandelion. Both the low and medium concentration extracts (8.3 g/L–12.5 g/L) showed an obvious promoting effect, while high concentrations exhibited distinct inhibiting effects. In conclusion, mint is the most suitable medicinal plant to be interplanted with fig trees for introduction into medicinal agroforestry systems.

Photosynthetic Characteristics and Yield Response of Isatis indigotica to Regulated Deficit Irrigation in a Cold and Arid Environment

Water resource scarcity is an important factor restricting the sustainable development of agriculture in Northwest China. Regulated deficit irrigation can conserve water while maintaining high crop yields. A field experiment was conducted to evaluate the effect of regulated deficit irrigation on the photosynthetic characteristics, yield, and water use efficiency of woad (Isatis indigotica) under mulched drip irrigation from 2017 to 2019 in a cold and arid area of the Hexi Oasis irrigation region, China. Sufficient water was supplied during the seedling stage. The control consisted of adequate water supplied during the other growth stages, whereas mild, moderate, and severe water deficits were imposed during the vegetative growth period, and a mild and moderate water deficit was imposed during the fleshy root growth stage. A mild water deficit was imposed during the fleshy root maturity period. The results showed that the net photosynthetic rate, transpiration rate, and stomatal conductance under moderate and severe water deficit were significantly (p < 0.05) decreased compared with the control, respectively, during the vegetative growth period. The economic yield of mild water deficit during the vegetative growth and mild water deficit during the vegetative growth and fleshy root growth did not differ significantly (p > 0.05) from that of the control. Other treatments caused a 6.74–17.74% reduction in the economic yield of woad. The water use efficiency and irrigation water use efficiency were the highest in the mild water deficit during the vegetative growth period and the fleshy root growth period. Therefore, the application of a continuous mild deficit from the vegetative growth stage to the fleshy root growth stage with sufficient water supplied during other growth periods is recommended as the optimal irrigation regime for maximum yield, water use efficiency, and water-saving of woad.

Identification of key gene networks controlling vernalization development characteristics of Isatis indigotica by full-length transcriptomes and gene expression profiles

Isatis indigotica Fort., as a common Chinese medicinal raw material, will lose its medicinal value if it blooms early, so it is highly valuable to clarify the induction mechanism of the vernalization of I. indigotica at low temperature. In this study, the concentrations of soluble sugar, proline, glutathione and zeatin in two germplasms of I. indigotica with different degrees of low temperature tolerance (Y1 and Y2) were determined at 10 days, 20 days and 30 days of low-temperature treatment, and the full-length transcriptome of 24 samples was sequenced by Nanopore sequencing with Oxford Nanopore Technologies (ONT). After that, the data of transcripts involved in the vernalization of I. indigotica at low temperature were obtained, and these transcripts were identified using weighted gene co-expression network analysis (WGCNA). The results revealed the massive accumulation of soluble sugar and proline in Y1 and Y2 after low temperature induction. A total of 18,385 new transcripts, 6168 transcription factors and 470 lncRNAs were obtained. Differential expression analysis showed that gibberellin, flavonoids, fatty acids and some processes related to low temperature response were significantly enriched. Eight key transcripts were identified by WGCNA, among which ONT.14640.1, ONT.9119.1, ONT.13080.2 and ONT.16007.1 encodes a flavonoid transporter, 9-cis-epoxycarotenoid dioxygenase 3 (NCED3), growth factor gene and L-aspartate oxidase in plants, respectively. It indicated that secondary metabolites such as hormones and flavonoids play an important role in the vernalization of I. indigotica. qRT-PCR proved the reliability of transcriptome results. These results provide important insights on the low-temperature vernalization of I. indigotica, and provide a research basis for analyzing the vernalization mechanism of I. indigotica.

Cover Image:

Cover Caption: Guan et al. (Page 2058‐2074) identified glucobrassicin as an anti‐microtubule compound from roots of Northern Ban Lan (Isatis tinctoria). The cover shows a photograph of Isatis tinctoria plant and GFP‐labelled microtubules (green) was eliminated by glucobrassicin in Tobacco BY‐2 cells.

Effects of perlite, grass charcoal and vermiculite on root growth of isatis (Isatis tinctoria L. Woad) and soil nutrient migration

Effects of perlite, vermiculite, and grass charcoal on root growth of Isatis and soil nutrients migration were studied, and the plant growth indicators and some soil properties of upper (0~5cm) and lower layer (5~15cm) were analyzed. The experiment treatments were loess (CK), loess : perlite = 3 : 1(A), loess : grass charcoal = 3 : 1(B), loess : vermiculite = 3 : 1(C), loess: perlite: grass charcoal = 6 : 1 : 1(A1), loess: perlite: vermiculite = 6 : 1 : 1(B1), loess: grass charcoal: vermiculite = 6 : 1 : 1(C1). The results showed that soil pH of vermiculite-containing treatments in the upper layer and grass charcoal treatment in the lower layer decreased significantly. Before planting, the available potassium and phosphorus of the upper layer were significantly higher than those of the lower layer, and soil organic matter (SOM) was slightly higher than that of the lower layer. After planting, the decrease of available potassium in the upper layer was less than that of the lower layer, and the decrease of available phosphorus and increase of SOM in the upper layer were slightly higher than that of the lower layer. In sum, it is preferred to choose B (loess : grass charcoal = 3 : 1) and A1 (loess: perlite: grass charcoal = 6 : 1 : 1) to improve the soil nutrient and utilization efficiency. Bangladesh J. Bot. 50(3): 947-954, 2021 (September) Special

Growth-promoting effects of dark septate endophytes on the non-mycorrhizal plant Isatis indigotica under different water conditions

Growth-promoting effects of dark septate endophytes on the non-mycorrhizal plant Isatis indigotica under different water conditions

Effects of Two Polysaccharides From Traditional Chinese Medicines on Rat Immune Function.

: The aim is to study the immune function effect of two polysaccharides extracted from traditional Chinese herbs on rats. Ultrasonic-assisted extraction was used to extract the polysaccharide from traditional Chinese medicines. MTT assay was used to determine the effects of two polysaccharides on the conversion of pig peripheral T lymphocytes. For this, 24 Sprague–Dawley rats were selected for the clinical trial and divided into groups B (blank), CK (cyclophosphamide inhibitory control), AP (angelica polysaccharide), and RIP (radix isatidis polysaccharide). Except for group B, other groups can induce the immunodeficiency by using cyclophosphamide. Rats of the AP and RIP groups were given gavage of 1 mL of AP and RIP. The blood was sampled from the eyeball on days 0, 7, 14, 21, 28, and 35, respectively, to determine immune cells, IgG and IgM of immunoglobulin, body weight, and spleen index.Results: The average content of AP and RIP was 51.27 and 14.8%, and the extraction rate was 75.23 and 60.94%. The maximum stimulation index was 1.407 when the concentration of AP was 8,000 μg mL−1 and 1.5 when the concentration of RIP was 125 μg mL−1. Both kinds of polysaccharides can alleviate the decline of white blood cells, lymphocytes, monocytes, neutrophils, and serum IgG and IgM caused by cyclophosphamide. The two polysaccharides can regulate the rapid recovery of weight in immunosuppressed rats and increase the spleen index of immunosuppressed SD rats. The polysaccharides from the two traditional Chinese medicines can alleviate the immunosuppression caused by cyclophosphamide and promote the immune function of the body, which can be used as raw material resources of new veterinary medicine.

Anti-microtubule activity of the traditional Chinese medicine herb Northern Ban Lan (Isatis tinctoria) leads to glucobrassicin.

Traditional Chinese medicine (TCM) belongs to the most elaborate and extensive systems of plant-based healing. The herb Northern Ban Lan (Isatis tinctoria) is famous for its antiviral and anti-inflammatory activity. Although numerous components isolated from I. tinctoria have been characterized so far, their modes of action have remained unclear. Here, we show that extracts from I. tinctoria exert anti-microtubular activity. Using time-lapse microscopy in living tobacco BY-2 (Nicotiana tabacum L. cv Bright Yellow 2) cells expressing green fluorescent protein-tubulin, we use activity-guided fractionation to screen out the biologically active compounds of I. tinctoria. Among 54 fractions obtained from either leaves or roots of I. tinctoria by methanol (MeOH/H2 O 8:2), or ethyl acetate extraction, one specific methanolic root fraction was selected, because it efficiently and rapidly eliminated microtubules. By combination of further purification with ultra-high-performance liquid chromatography and high-resolution tandem mass spectrometry most of the bioactivity could be assigned to the glucosinolate compound glucobrassicin. Glucobrassicin can also affect microtubules and induce apoptosis in HeLa cells. In the light of these findings, the antiviral activity of Northern Ban Lan is discussed in the context of microtubules being hijacked by many viral pathogens for cell-to-cell spread.

Stabilization of natural and synthetic indigo on nanocellulose network - Towards bioactive materials and facile dyeing processes

Synthetic dyes are vastly used for colouring numerous materials, although the adverse effects on environment are well recognized. In addition to developing the existing dyeing technologies more efficient and cleaner, the valorisation of natural dyes can enhance the sustainable development of dyeing industry. Natural indigo, derived from Isatis tinctoria, is a bio-based alternative for indigo produced via chemical synthesis routes. Owing to the insoluble character of indigo pigment, the dye requires conversion into soluble leucoindigo form prior to dyeing, which is often accomplished by using harsh sodium dithionite vat technique. During the processing from plant to dye attached on a fabric, indigo is transferred from the soluble leucoindigo form to the oxidized insoluble indigo and once more back to leucoindigo. Additionally, the oxidation is difficult to control and with traditional vat technique maintaining the leucoindigo through the dyeing often requires adding more reducing agent chemicals. Maintaining the soluble form throughout the process would enable lower number of processing steps and reduce the use of harmful chemical agents. In the present study, the stabilization of leucoindigo on nanocellulose matrix carrier was investigated with spectroscopic and photophysical methods. According to the results, leucoindigo was successfully stabilized on nanocellulose suspension, most likely due to the limited rate of oxygen diffusion into the viscous medium. Visual observations revealed that the leuco-form was retained even longer with natural indigo than synthetic indigo. This enhanced stability was attributed to the presence of radical scavenging species in natural indigo since the synthetic indigo did not show notable antioxidant properties. Given the promising results the paste formulation was demonstrated to be applicable for creating patterns on cotton using a screen-printing technique. Since the leucoindigo was stabilized on nanocellulose carrier, the need for re-reduction prior to dyeing was avoided and the amount of harmful reducing chemicals was reduced. These findings also show that the characteristics of natural dyes that are often considered disadvantageous compared to synthetic dyestuff, i.e. presence of co-products in the mixture, can however, create more value to the dyed material through new functionalities.

Colour stability of wood coatings pigmented with natural indigo from Isatis tinctoria after accelerated weathering

AbstractAn experimental study of artificial weathering of two outdoor coatings pigmented with natural indigo from Isatis tinctoria was carried out. The two coatings, natural indigo‐pigmented boat varnish and outdoor paint, were exposed to cycles of ultraviolet light, water condensation and water spray for 40 days in a QUV test chamber. Synthetic indigo and commercial ultramarine pigment were employed as reference materials. The effects of artificial weathering on the coatings were evaluated with CIELab parameters, as well as visual inspection. Coating and pigment stability during artificial weathering were studied with reflectance spectrometry, hyperspectral imaging technology and Fourier Transform–infrared spectroscopy. According to the results, coatings with indigo proved to be more resistant to visual and spectral changes than unpigmented or commercially pigmented coatings.

Immunomodulatory effects of Radix isatidis polysaccharides in vitro and in vivo.

Radix isatidis (R. isatidis) is a commonly used traditional Chinese herbal medicine, which has been used for thousands of years in China and is believed to have the pharmacological properties of heat-clearing and detoxification. Heat-clearing and detoxification are theories of traditional Chinese medicine meaning that R. isatidis could treat febrile disease by clearing heat and reducing swelling. Polysaccharides isolated from R. isatidis by water extraction and alcohol precipitation have exhibited numerous biological activities, including antiviral and immunomodulatory effects. The present study was performed to investigate the immunomodulatory effects of water-soluble R. isatidis polysaccharides (RIPs) on RAW264.7 macrophages and murine splenocytes, and attempt to preliminarily identify the mechanism of immunomodulation. In vitro, RIPs had a low cytotoxicity, as shown by CellTiter 96® AQueous One Solution Cell Proliferation Assay. RAW264.7 cells treated with different concentrations of RIP displayed different morphological changes, from a round shape and aggregation to polygonal shape and dispersion in a dose-dependent manner. In the 5 mg/ml RIP-treated group, the changes of morphology were as same as the lipopolysaccharide-treated group. RIP also significantly enhanced the release of nitric oxide as shown by Griess method, and the secretion of TNF-α and IL-6 in RAW264.7 cells was confirmed by ELISA assay. Western blotting revealed a significant increase of toll-like receptor-4 (TLR-4) in RIP-treated RAW264.7, suggesting that TLR-4 may be associated with the immunomodulatory mechanism of RIP. Animal experiments also demonstrated through ELISA assays a significant increase in IFN-γ and IL-10 levels after the splenocytes of RIP-immunized mice were stimulated by inactivated herpes simplex virus type 2. The immune function of RIP-immunized mice was improved. The present study suggested that RIP could be potentially used as a novel immunomodulator.

Integrative analyses of targeted metabolome and transcriptome of Isatidis Radix autotetraploids highlighted key polyploidization-responsive regulators

BackgroundIsatidis Radix, the root of Isatis indigotica Fort. (Chinese woad) can produce a variety of efficacious compound with medicinal properties. The tetraploid I. indigotica plants exhibit superior phenotypic traits, such as greater yield, higher bioactive compounds accumulation and enhanced stress tolerance. In this study, a comparative transcriptomic and metabolomic study on Isatidis Radix autotetraploid and its progenitor was performed.ResultsThrough the targeted metabolic profiling, 283 metabolites were identified in Isatidis Radix, and 70 polyploidization-altered metabolites were obtained. Moreover, the production of lignans was significantly increased post polyploidization, which implied that polyploidization-modulated changes in lignan biosynthesis. Regarding the transcriptomic shift, 2065 differentially expressed genes (DEGs) were identified as being polyploidy-responsive genes, and the polyploidization-altered DEGs were enriched in phenylpropanoid biosynthesis and plant hormone signal transduction. The further integrative analysis of polyploidy-responsive metabolome and transcriptome showed that 1584 DEGs were highly correlated with the 70 polyploidization-altered metabolites, and the transcriptional factors TFs-lignans network highlighted 10 polyploidy-altered TFs and 17 fluctuated phenylpropanoid pathway compounds.ConclusionsThese results collectively indicated that polyploidization contributed to the high content of active compounds in autotetraploid roots, and the gene–lignan pathway network analysis highlighted polyploidy–responsive key functional genes and regulators.

Discovery of alkaloids from the leaves of Isatis indigotica Fortune with neuroprotective activity.

Seven alkaloids including five undescribed ones (1a/1b, 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 1a, 1b, 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 H2O2-induced injury in SH-SY5Y 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 1a and 1b were measured.

Effects of water and nitrogen coupling on the photosynthetic characteristics, yield, and quality of Isatis indigotica

Isatis indigotica is a commercial medicinal crop that is widely cultivated with high water and nutrient application, in the arid areas of northwest China. Rational irrigation and nitrogen application are key factors for successful crop management. The objective of this study was to determine the effect of water and nitrogen coupling on the photosynthetic characteristics, yield, and quality of Isatis indigotica produced in northwestern China. Field trials were conducted for 2 consecutive years at an irrigation test station. Data on photosynthetic parameters, yield, and quality were collected from individual Isatis indigotica for each treatment during 2018–2019. The application of nitrogen significantly increased photosynthetic rates and yield under the same irrigation conditions. However, the yields were reduced in the excess water treatments (W3N1 and W3N2) and in the excess nitrogen treatments (W1N3, W2N3, and W3N3) in contrast to the optimum W2N2 treatment. Moreover, the quality indicators of the W2N2 treatment decreased compared with CK, which was due to water stress and more photoassimilates being available to the roots, but the effective quality index value could be effectively improved by greatly increasing the yield.

Anti-Wrinkle Effect of Isatis indigotica Leaf Extract: Evaluation of Antioxidant, Anti-Inflammation, and Clinical Activity.

Isatis indigotica leaf is an oriental herbal medicine that has been known for various pharmacological effects. However, its anti-wrinkle activity has not been fully evaluated. Therefore, we evaluated the anti-wrinkle effect of I. indigotica leaf extract on human skin. The purified extract inhibited 85.4% of 2,2-diphenyl-1-1picrylhydrazyl and 72.2% of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radicals at a concentration of 1 mg/mL. Nitrite production was reduced by 30% after treatment with 50 μg/mL of extract. Three fractions from the extract downregulated the mRNA expression of matrix metalloproteinase-1 and -3 and upregulated the expression of interleukin 4. Among the three fractions, fraction 2 exhibited the highest activity. The major component of the extract was identified as 3,4,5-trimethoxycinnamic acid by liquid chromatography coupled with mass spectrometry. Molecular docking was conducted to predict the binding mechanism of 3,4,5-trimethoxycinnamic with matrix metalloproteinase-1 and -3, and their binding energies were −5.20 and −4.89 kcal/mol, respectively. In a clinical trial, five roughness values of visiometer and visual score were significantly reduced in treated groups compared with the placebo group after 8 weeks. I. indigotica leaf extract inhibits wrinkle formation, and could be a potential anti-wrinkle agent. This is the first clinical trial demonstrating its anti-wrinkle activity.

Network pharmacology study on the mechanism of the Chinese medicine Radix Isatidis (Banlangen) for COVID-19.

Radix Isatidis (Banlangen) is a well-known traditional Chinese medicine for the treatment of different diseases and prevention of many body disorders. Besides, it also plays a pivotal role in novel coronavirus pneumonia, coronavirus disease 2019 (COVID-19). However, few researchers know its active ingredients and mechanism of action for COVID-19. To find whether Banlangen has a pharmacological effect on COVID-19. In this research, we systematically analyze Banlangen and COVID-19 through network pharmacology technology. A total of 33 active ingredients in Banlangen, 92 targets of the active ingredients, and 259 appropriate targets of COVID-19 were obtained, with 11 common targets. The analysis of the biological process of gene ontology and the enrichment of Kyoto Encyclopedia of Genes and Genomes signaling pathway suggests that Banlangen participated in the biological processes of protein phosphatase binding, tetrapyrrole binding, the apoptotic process involving cysteine-type endopeptidase activity, etc. The COVID-19 may be treated by regulating advanced glycation end products/a receptor for advanced glycation end products signaling pathway, interleukin-17 signaling pathway, tumor necrosis factor signaling pathway, sphingolipid signaling pathway, and p53 signaling pathway. Banlangen has a potential pharmacological effect on COVID-19, which has the value of further exploration in the following experiment and clinical application.

A Network Pharmacology-Based Investigation to the Pharmacodynamic Material Basis and Mechanisms of the Anti-Inflammatory and Anti-Viral Effect of Isatis indigotica.

PurposeIsatis indigotica (Ii) is a cruciferous herb that is widely distributed in China, and its roots and leaves have been used in two renowned antipyretic detoxicate crude drugs in Chinese Pharmacopoeia, Radix (R) and Folium (F) Isatidis. However, the pharmacodynamic material basis and underlying mechanisms of the herbal efficacy remained to be elucidated.MethodsUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was adopted for the chemical profiling of R and F Isatidis. The active ingredients were screened out through the prediction of gastrointestinal absorption and druglikeness analysis using SwissADME. A herb-ingredient-target network was constructed through target prediction of the herbal active ingredients and anti-inflammation or anti-viral properties, followed by protein–protein interaction analysis. Then, the potential relevant signaling pathways were predicted by pathway enrichment. Finally, for verification, RAW 264.7 cell line was adopted to examine the anti-inflammatory and anti-viral activities of 6 representative ingredients in Ii.ResultsSeventy-three compounds have been identified from Ii through UPLC-Q-TOF-MS. A total of 17 potential active ingredients were screened through pharmacokinetics and drug-likeness evaluation using SwissADME. It was shown that key targets might include TNF, AKT1, SRC, IL2, CASP9, and CASP3 in our herb-ingredient-target network, and isovitexin, a flavonoid, tended to participate in the inflammatory response, indoles were more likely to affect the cell proliferation processes, and lignans might have a broader affinity to key targets than the other active ingredients, such as regulating immune system (targeting IL-2) and PI3K-Akt signaling pathway. In vitro, indigo and secoisolariciresinol diglucoside markedly reduced TNF-α expression in Poly (I: C)-incubated cells. Isovitexin significantly inhibited TNF-α expression, and isatin treatment markedly reduced IL-1β expression in LPS-incubated cells.ConclusionAs the pharmacodynamics material basis of Ii, indoles, lignans, and flavonoids are believed to confer beneficial properties through various cellular aspects with multiple signaling pathways involved.