Correlation analysis between the HPLC fingerprints and in vitro antibacterial activity of ethyl acetate extracts from Radix isatidis

Aim To study the correlation between the HPLC fingerprints and in vitro antibacterial activities of EtOAc extracts of Radix isatidis from various sources. Methods Ten batches of Radix isatidis EtOAc extracts were analyzed with HPLC and the fingerprints were established. The influence of EtOAc extracts on the thermogenic curve of growth of Escherchia coli was obtained by microcalorimetry. The chemical differences of EtOAc extracts of Radix isatidis from various sources in the HPLC fingerprints were probed with hierarchical clustering analysis and similarity analysis. The correlation between the HPLC fingerprints and in vitro antibacterial activities was analyzed with multivafiant correlation analysis. Results Close correlation existed between the HPLC fingerprints and in vitro antibacterial activities of EtOAc extracts of Radix isatidis. Conlusion The combination of HPLC fingerprints and antibacterial activities can be used to discover principle components of Radix isatidis on bioactivity.

Antiviral Activity of the Effective Monomers from Folium Isatidis Against Influenza Virus in Vivo

In order to evaluate the anti-influenza virus activity of the effective monomer from Folium Isatidis (FI) in vivo,we established mice model with viral pneumonia and divided them into 3 different dose groups,then observed their lung indexes,pulmonary pathological changes,pulmonary virus hemagglitination titers,living time and death rates.The results showed that the monomer could reduce the pulmonary index from 2.64 to 1.93,1.63 and 1.40 (P<0.01) and decrease the hemagglitination titer from 1.15 to 0.84,0.70 and 0.59 (P<0.01).In addition,different groups of FI could significantly lessen the mortality rate from 100% to 30%,25% and 15%,and prolong the living time from 5.1d to 6.5d,8.4d and 8.9d respectively(P<0.01).The high dose (75 mg/kg/d) has the similar effect with 100 mg/kg/d dose of virazole(P>0.05),and more effective than 200 mg/kg/d dose of antiviral liquor (P<0.05).

Determination of Five Organic Acids in Radix Isatidis by Column Partition Chromatography and Capillary Zone Electrophoresis柱分配色谱-毛细管区带电泳联用分析板蓝根中五种微量有机酸(英文)

Aim To determine five organic acids in Radix Isatidis . Method The extraction method and the column partition chromatographic conditions were studied. Then a capillary zone electrophoretic method was set up for the determination. Results The linear ranges of quinazolinone acid, n anthranilic acid, benzoic acid, salicylic acid, and syringic acid were 5 52-92 0 μg·mL -1 , 5 12-102 μg·mL -1 , 2 28-84 4 μg·mL-1 , 4 78-159 μg·mL -1 , and 1 74-87 0 μg·mL -1 respectively. Conclusion The established method is accurate and simple.

Effect of Radix Isatidis on the Expression of Moesin mRNA Induced by LPS in the Tissues of Mice

To investigate the effect of the anti-endotoxic part of Radix Isatidis on the expression of moesin mRNA in murine tissues induced by lipopolysaccharide （LPS）, the sample solution of F0z2 part from Radix Isatidis was intrapefitoneally administered to experimental mice, and the lipopolysaccharide （LPS） were injected into the tail vein, and then the tissues of liver, kidney and spleen were colleted and cut into slices. The mRNA was detected by moesin mRNA hybridization in situ. The staining results were observed under microscope. It was found that moesin mRNA expression was increased in the tissues of liver, kidndy and spleen in mice treated with LPS, while in the mice pre-treated with F022 part from Radix Isatidis, the LPS-induced moesin mRNA expressions in these tissues were inhibited in a dose-dependant manner. Our study showed that F022 part from Radix Isatidis can inhibit the LPS-induced expression of moesin mRNA in the tissues of liver, kidney and spleen in mice.

Particle Sizes and Antibacterial Activity of Radix Astragalus and Radix Isatidis Ultrafine Powders

Using 9200 laser particle size analyzer and KYKY-2800 scanning electron microscope, particle sizes and cellular morphology of Radix Astragalus and Radix Isatidis ultrafine powders were observed. According to the results, the particle size of 89. 1 % of Radix Astragalus ultrafine powders ranged from 1.729 [xm to 44.938 ｜xm, Z ）50 =4.368 ｜xm; the particle size of 93.411% of Radix Isatidis ultrafine powders ranged from 1.510 [xm to 44.938 ｜xm, Z ）50 = 8 .7 2 6 [xm. Radix As-tragalus and Radix Isatidis ultrafine powders were pulverized completely without intact cellular morphology. The antibacterial activity of Radix Astragalus and Radix Isatidis ultrafine powders against chicken-derived E. coli （078） was investigated. The results indicated that Radix Astragalus and Radix Isatidis ultrafine powders exhibited higher antibacterial activity against chicken-derived E. coli （078 ） compared with the corresponding coarse powders. This study laid a solid foundation for the development and application of Chinese medicine ultrafine powder preparations.

Screening of Anti-endotoxin Components from Radix Isatidis

To find a component with the strongest anti-endotoxin effect from the four components （F021 , F022, F023, and F024 ） of chloroform extract （F02） of Radix Isatidis, the protective effects of the five components on endotoxin-challenged mice and their effects on the production of TNFα and IL-6 by macrophages from the endotoxin-challenged mice were compared. It was found that the five components all had protective effects on endotoxin-challenged mice in terms of mortality. The mortalities of the mice challenged by 2.42 mg/kg endotoxin were 30 %, 50 %, 20 %, 50 % and 60 0% after treatment with F02, F021 , F022 , F023 , and F024. The 5 components all had inhibitory effects on the production of TNFα and IL-6 by macrophages from the mice. At a concentration of LPS of 50 ng/mL, the inhibitory rates of F02, F021 1, F022, F023, and F024 for the production of TNFα and IL-6 were 76.54 %, 30. 86 %, 78.60 %, 36. 63 %, 2.06 % and 75.85 %, 18.45 %, 77. 68 %, 24. 41 %, 3. 64% and at the dose of 100 ng/mL, the inhibitory rates were 49. 65 %, 16.86%, 66.97 %, 13. 39 %, 7.16 % and 59.78 %, 1.28 %, 61.86 %, 2.08 %, 1.44 %, respectively. It is concluded that the component F022 of Radix Isatidis has the strongest anti-endotoxin effect.

The Influence of Polysaccharides Extracted from Radix Isatidis on Immunity

The Radix Isatidis polysaccharides (IIP) injected ip at the dose of 50mg/kg daily for seven days increased the spleen index. the total numbero of leukocytes and lymphocytes,the percentage of ANAE+ lymphocytes and the intensity of delayed type hyperoensitivity in normal mice as well as in immunosuppressed mice.Moreover,IIP could improve the phagocytic function of reticuloendothelial system,promote the yield of hemolysin,and increase the NK activity in norwal mice at the same dose.

Anti-Endotoxic Effects of Syringic Acid of Radix Isatidis

The anti-endotoxic effect of syringic acid (SA) isolated from Radix Isatidis (Banlangen, BLG) was studied. SA was extracted and isolated from BLG and diluted into 1 % solution. The content of SA-pretreated endotoxin (ET) was quantitatively determined using Limulus test. The ability of fever induction of ET pretreated with SA was measured using endotoxin-induced fever test in rabbits. The LPS-induced death in mice pretreated with and without SA was compared. Results showed that after pretreatment with SA, 83. 16 % of ET was destroyed, the ET-induced fever in rabbits relieved markedly and the LPS-induced death rate in mice dropped from 68 % to 20 %. It was concluded that SA isolated from BLG had anti-endotoxic effects.

Associated Bacteriostasis Research of Coptidis Rhizoma-Folium Isatidis and Coptidis Rhizoma-Flos Poprli against Escherichia coli O2

[Objective] The paper was to explore the combined inhibitory effect of rhizoma coptidis-folium isatidis and rhizoma coptidis-flos poprli against Escherichia coli O2.[Method] Contrast test of single and associated bacteriostasis against known serotype E. coli O2 was conducted using microcheckerboard method.[Result] The MIC of rhizoma coptidis, folium isatidis and flos poprli were 1/8 extracting liquid, 1/8 extracting liquid and 1/2 extracting liquid, respectively. When combined with folium isatidis or flos poprli, the MIC of rhizoma coptidis was 1/8 extracting liq-uid or 1/16 extracting liquid compared with single use. When combined with rhizoma coptidis, the MIC of folium isatidis and flos poprli were 1/8 extracting liquid and 1/16 extracting liquid.[Conclusion] When rhizoma coptidis was combined with folium isatidis or flos poprli, the FIC values were 2 and 0.625, performing independent action and additive effect, respectively.

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

Background:To predict the moisture ratio of Radix isatidis extract during drying.Methods:Artificial neural networks were designed using the MATLAB neural network toolbox to produce a moisture ratio prediction model of Radix isatidis extract during hot air drying and vacuum drying,where regression values and mean squared error were used as evaluation indexes to optimize the number of hidden layer nodes and determine the topological structure of artificial neural networks model.In addition,the drying curves for the different drying parameters were analyzed.Results:The optimal topological structure of the moisture ratio prediction model for hot air drying and vacuum drying of Radix isatidis extract were“4-9-1”and“5-9-1”respectively,and the regression values between the predicted value and the experimental value is close to 1.This indicates that it has a high prediction accuracy.The moisture ratio gradually decreases with an increase in the drying time,reducing the loading,initial moisture content,increasing the temperature,and pressure can shorten the drying time and improve the drying efficiency.Conclusion:Artificial neural networks technology has the advantages of rapid and accurate prediction,and can provide a theoretical basis and technical support for online prediction during the drying process of the extract.

Influence of Radix Isatidis on the Endotoxin-induced Release of TNF-α and IL-8 from HL-60 Cells

Summary： The effects of active antiendotoxin chemical fraction isolated from Radix Isatidis （fraction D） on TNF-α and IL-8 secretion in HL-60 cells induced by lipopolysaccharide （LPS） were studied. The appropriate densities of cell suspension and fraction D solution were determined by MTT colorimetric method. Fraction D and LPS were added to HL-60 cell suspension with three different methods respectively. The contents of TNF-α and IL-8 in the cultured supernatant induced by LPS were detected by using ELISA method. The results showed that the absorbance （A） was directly proportional to the number of cells and the linearity was good in the range from 0.25×10^5 to 2×10^5 cell/mL cell suspension. The fraction D significantly inhibited the oversecretion of TNF-α and IL-8 in HL-60 cells induced by LPS at the concentration of 7.812 mg/mL which had no cytotoxicity. It was indicated that the antiendotoxin mechanism of the active fraction from Radix Isatidis was contributed to the inhibition of the oversecretion of cvtokines induced bv LPS.

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

Background:Drying is a necessary component of traditional Chinese medicine extracts.The heating principle of microwave vacuum drying is different from that of the conventional heat method.However,at present,there is paucity of information on the drying process of traditional Chinese medicine extract by microwave vacuum drying,and the results of such process are unclear.Methods:To study the dynamic changes in the chemical characteristics of microwave vacuum drying under different drying conditions,ultrahigh-performance liquid chromatography fingerprint profiles were established using Radix isatidis extract as a model drug and analyzed using similarity analysis,partial least squares-discriminant analysis,and semi-quantitative analysis.In addition,a backpropagation artificial neural network model was developed to predict the moisture ratio of the drying process.Results:Qualitative results showed that the similarity between different drying conditions was greater than 0.95,and 2 amino acid components(peaks 5 and 6)affected by process fluctuations were screened out.The quantitative results showed that the mass concentration of component 1 fluctuated after drying,while that of component 2 increased.The optimal backpropagation artificial neural network model structure used to predict the moisture ratio was 5-4-1,with regression and mean squared error values of 0.996 and 0.0003,respectively,after training,which were well fitted and had a strong approximation ability.Conclusion:Upon comparison of fingerprints and the evaluation of statistical methods,common components of Radix isatidis extract had little variation under different drying conditions,and the selected components provided a reference for the establishment of process evaluation indexes.The establishment of backpropagation artificial neural network provides a theoretical basis for the application of microwave vacuum drying technology and online monitoring of moisture ratio.

Mechanism of Radix isatidis in influencing pulmonary inflammation in tuberculosis rats by TLR4-NF-κB pathway

Objective:To investigate the mechanism of the effects of Radix isatidis on lung inflammation in tuberculosis rats through TLR4-NF-κB pathway.Methods:Forty-five SD rats were randomly divided into three groups(n=15 in each group):control group,model group,and isatis root polysaccharide group.Through the tail vein injection of H37RV Mycobacterium tuberculosis model,the rats in the isatis root polysaccharide group received the intervention of Radix isatidis polysaccharides on the second day of infection.The serum inflammatory factors and the expression levels of TLR4-NF-κB pathway and inflammatory factors in lung tissue of each group were detected and compared.Results:A large number of Mycobacterium tuberculosis infection in the lung tissue of the model group,and clustered outside the cell.In the isatis root polysaccharide group,the CFU level was significantly lower compared with the model group,and the number of Mycobacterium tuberculosis was less than that of the model group(P<0.05),mainly distributed in tuberculous nodules and giant cells.The level of serum inflammatory factors in the model group was significantly higher than that in the control group(P<0.05).The levels of IL-1β,IL-6 and IFN-γin the serum of the isatis root polysaccharide group were significantly lower than those in the model group and significantly higher than those in the control group(P<0.05).The TLR4-NF-κB pathway and inflammatory factor mRNA levels in the lung tissue of the model group were significantly higher than those of the control group(P<0.05).The levels of TLR4,NF-κB,IL-1β,IL-6 and IFN-γin the lung tissue of the isatis root polysaccharide group were significantly lower than those of the model group and significantly higher than those of the control group(P<0.05).The expression level of TLR4-NF-κB pathway was significantly up-regulated in the lung tissue of model rats(P<0.05).The levels of TLR4 and NF-κB in the lung tissue of the isatis root polysaccharide group were significantly lower than those of the model group and significantly higher than those of the control group(P<0.05).Conclusion:Radix isatidis can alleviate lung inflammation caused by Mycobacterium tuberculosis infection by regulating TLR4-NF-κB pathway.

超声波辅助提取大青叶中总黄酮及其抗氧化性研究

以大青叶(Folium Isatidis)为原料,采用单因素试验法,研究了乙醇浓度、超声时间、料液比和提取次数等因素对大青叶中总黄酮提取率的影响,得到大青叶中总黄酮的最适提取工艺条件为乙醇浓度70%,超声时间30 min,料液比(g/mL)1∶25和提取次数2次。该试验结果可为大青叶资源的进一步开发利用提供参考。

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