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Four alkaloids, previously identified in Isatis species, were tested for their inhibitory effect on histamine release. Whereas tryptanthrin, indirubin and deoxyvasicinone did not inhibit histamine release, the effect of indolin-2-one exceeded that of the mast cell stabilizing drug disodium chromoglycate.
The aim of the study was to evaluate the effects of two kinds of Chinese medicinal herbs, Isatis tinctoria L (ITL) and Daphne giraldii Nitsche (DGN), on a rat model of chronic Pseudomonas aeruginosa lung infection mimicking cystic fibrosis (CF). Compared to the control group, both drugs were able to reduce the incidence of lung abscess (p < 0.05) and to decrease the severity of the macroscopic pathology in lungs (p < 0.05). In the great majority of the rats, the herbs altered the inflammatory response in the lungs from an acute type inflammation, dominated by polymorphonuclear leukocytes (PMN), to a chronic type inflammation, dominated by mononuclear leukocytes (MN). DGN also improved the clearance of P. aeruginosa from the lungs (p < 0.03) compared with the control group. There were no significant differences between the control group and the two herbal groups with regard to serum IgG and IgA anti-P. aeruginosa sonicate antibodies. However, the IgM concentration in the ITL group was significantly lower than in the control group (p < 0.03). These results suggest that the two medicinal herbs might be helpful to CF patients with chronic P. aeruginosa lung infection, DGN being the most favorable.
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.
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 TNFalpha 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 % after treatment with F02, F021, F022, F023, and F024. The 5 components all had inhibitory effects on the production of TNFalpha 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 TNFalpha 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 Y, 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 effects of active antiendotoxin chemical fraction isolated from Radix Isatidis (fraction D) on TNF-alpha 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-alpha 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 x 10(5) to 2 x 10(5) cell/mL cell suspension. The fraction D significantly inhibited the oversecretion of TNF-alpha 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 cytokines induced by LPS.
AIM: To study the anti-endotoxic effect of o-aminobenzoic acid (OABA) isolated from Banlangen(BLG). METHODS: OABA was extracted and isolated from BLG and diluted into 0.5% solution. The concentration of endotoxin (ET) pretreated with OABA was quantitatively detected using Limulus test. The inhibition of ET-induced fever by OABA was measured in rabbits. The rates of lipopolysaccharides (LPS)-induced death in mice pretreated with or without OABA were then compared. The influence of OABA on the release of TNF-alpha and NO from macrophages induced by LPS was examined in mice. RESULTS: After pretreatment with OABA, 84.4% of ET was destroyed. The ET-induced fever in rabbits decreased significantly and the rate of LPS-induced death in mice dropped from 70% to 20%. The release of TNF-alpha and NO induced by LPS in mice was inhibited dose-dependently when the concentration of OABA was between 0.125% and 0.5%. CONCLUSION: OABA isolated from BLG has an anti-endotoxic effect.
Cycloanthranilylproline (1) and its derivatives (2--4) were isolated from field-collected fruit-bodies of a myxomycete Fuligo candida and their structures were elucidated by spectral data. Compound 4, which was contained in the water-soluble fraction of the extract of this myxomycete, was unstable and quite susceptible to decarboxylation to yield compound 2, which was a major constituent of the EtOAc-soluble fraction of this extract.
OBJECTIVE: To research into the cause of distinction in antiendotoxic activation of Radix Isatidis from different cultivated populations. METHOD: 19 samples of Radix Isatidis were gathered. 12 portions of seeds were cultivated in the same field in Shanghai, and the selected and artificially bred autotetraploid seeds were planted in 5 different locations. All the samples were compared through antiendotoxic experiment. RESULTS: There are some marked distinction among the 19 samples from different populations. The distinction of the 12 samples cultivated in the same field didn't disappear. The activation of autotetraploid from 5 different locations didn't vary on the whole. CONCLUSION: Germplasm is the main factor to cause activation distinctions of Radix Isatidis, while environment is less important.
27 samples of indigowoad root and 5 samples of indigowoad leaf from different species and in different habital were collected. The chemical constituents were analyzed by HPLC, and ten original chemical features as a whole obtained. Meanwhile, extrinsic bacteriostasis experiments were carried out for each of the 32 samples. Canonical correlation analysis was applied to the intercorrelation between the chemical and the pharmacologic data, and five chemical features were found to be the effective constituents. In the end, the 32 samples were divided into five classes by the fuzzy clustering technique ISODATA(interachive self-organizing data analysis technique A).
OBJECTIVE: To study the anti-endotoxin activity and mechanism of F022 from Radix Isatidis. METHOD: The production of TNF-alpha and IL-6 of murine peritoneal macrophages stimulated by LPS was measured by ELISA. The temperature in rabbits was tested after i.v. administration of LPS. The lethality of BCG-primed mice was induced by LPS. RESULT: If F022 was added to macrophages culture simultaneously with LPS or 1 h before addition of LPS, production of TNF-alpha and IL-6 by macrophages was remarkably inhibited in vitro. F022 inhibited the fever induced by LPS in rabbits and protected BCG-primed mice from LPS induced lethality if given before administration of LPS. CONCLUSION: The anti-endotoxin effect of F022 may inhibit LPS binding to its receptor, and it may be a LPS receptor antagonist.
OBJECTIVE: To study the pharmacokinetics and bioavailability of syringic acid(SA) in rabbits by HPLC. METHOD: After the rabbits were injected SA injection via ear vein and peritoneal cavity, blood was collected also via ear vein at different time-point. Then the concentration of SA in blood was determined by HPLC and the concentration-time curve was mapped to simulate a compartment model. So the pharmacokinetic parameters were calculated out. Several days after this injection, the drug was administered to the rabbits via i.p. and using the same method, another concentration-time curve was obtained. By comparing the AUC of the two curves, bioavailability was obtained. RESULTS: The concentration-time curve of SA in rabbits after i.v. fitted in a two-compartment open model and the absolute bioavailability was 86.27%. CONCLUSION: The results were valuable for the clinical application of new anti-endotoxic drug from Radix Isatidis.
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OBJECTIVE: To investigate the difference of anticytomegaloviral activity of three kinds of traditional Chinese medicines which are the injections of Jinye Baidu, Radix Isatidis and Indigowoa in vitro. METHOD: The inhibitory activity of three traditional Chinese medicines against human cytomeglovirus (HCMV AD169) infected human embryo lung fibroblasts (HELF) was observed by cytopathic effect method (CPE) and MTT method in vitro. According their value of A, anticytomegaloviral activity has evaluated. RESULT: Experimental study in vitro showed that the 50% toxicity dose (TD50) of Jinye Detoxifying, Radix Isatidis root and Indigowoa were 20, 10.23, 20.23 g x L(-1) respectively; the 50% inhibitory concertration (IC50) were 5.65, 3, 5.71 g x L(-1) respectively; the therapeutic index (TI) were 3.54, 3.41 and 3.54 respectively. It suggested that three traditional Chinese medicines had anticytomeglovirus activity and their effect increased with their concentration. CONCLUSION: Three traditional Chinese medicines of the parenteral solution of Jinye Detoxifying, Radix Isatidis root and Indigowoa have antiviral activity when they are diluted in 1:200. They are safe and valuable drug for inhibiting cytomeglovirus infection.
OBJECTIVE: To evaluate the antiviral effect of different chemical fractions isolated from Folium Isatidis on herpes simplex virus type I (HSV- I). METHOD: Anti- HSV- I effects of Folium Isatidis were investigated in Hep-2 cell by adopting MTT colorimetric assay and observing cytopathic effect (CPE). Treatment index (TI) was used to evaluate the antiviral activity of different chemical fractions from Folium Isatidis. The antiviral mechanism of Folium Isatidis was investigated by changing the different ways of drug administration. RESULT: The fractions II approximately V from Folium Isatidis inactivate HSV- I directly. None of the chemical fractions had antiviral effects of adsorption-blocking. The chemical fractions except fraction III and fraction V inhibited the biological synthesis of HSV- I. The fraction IV significantly reduce death rate of mice infected with HSV- I. CONCLUSION: The fraction IV from Folium Isatidis has powerful anti-HSV-lI effect in vitro and in vivo.
OBJECTIVE: To observe the different extracts from Radix Isatidis on multiplication of mice lymphocytes. METHOD: Lymphocytes were separated and cultured. Immunological activities of different extracts from Radix Isatidis were studied by thermodynamics and the results were tested by the conventional pharmacological experiments. RESULT: The results showed that the water extract and residue had significant immunological effects while organic solvent extracts had immunological activity to some extent. CONCLUSION: The comparison of immunological activity among the extracts from Radix Isatidis were as follows: residue after extracting > general extract > nBuOH extract > EtOAc extract > CHCl3 extract > P E extract.
OBJECTIVE: To develop the chromatographic fingerprints of composite Folii Isatidis injection by HPLC. METHOD: The separation was performed on a Diamonsil C18 column with a mobile phase consisting of methanol-water-phosphoric acid as gradient eluent at the flow rate of 1.0 mL x min(-1). The UV detection was set at 254 nm. RESULT: A standard HPLC fingerprint procedure was developed for composite Folii Isatidis injection, with 20 common peaks and a similarity threshold of 9.0 established. CONCLUSION: This method was accurate, repeatable and useful for the quality control of composite Folii Isatidis injection.
OBJECTIVE: To develop a quantitative method for determination of the total organic acids and salicyclic acid in the extract of Radix Isatidis. METHOD: The total organic acids were determined by acid-base titration and the salicylic acid was determined by HPLC. RESULT: It was shown that contents of total organic acids and salicylic acid in the extract of Radix Isatidis were 13.0% and 0.22%, respectively. CONCLUSION: The method can control the quality of this extract effectively and accurately.
OBJECTIVE: To research the mechanism of the inhibition effects of BWE on cell attachment of influenza virus by capillary electrophoresis. METHOD: The morphologic difference of red cells after treating with BWE infected by influenza virus was detected with microscope, capillary electrophoresis and HA. RESULT: The pretreatment of the normal cells with BWE inhibited the attachment of influenza to the cells, while no meaningful inhibition was observed when influenza virus was pretreated before being inoculated to cells. CONCLUSION: The results indicate that the inhibition effects of BWE on cell attachment of influenza virus may be an important mechanism of anti-influenza activity of Radix Isatidis Extracts.