Anti-arthritic activity of a lipophilic woad (Isatis tinctoria) extract

A dichloromethane extract of Isatis tinctoria was tested in the adjuvant-induced arthritis model in rats. The extract (150 mg/kg p. o.) leads to a significant reduction of paw oedema. Radiographic, histological and clinical assessment confirmed reduced damage of cartilage and signs of inflammatory response in comparison to untreated control. No significant differences were observed in the tissular levels of cyclooxygenases 1 and -2, and of inducible nitric oxide synthase in Isatis treated and untreated animals. High dose treatment with Isatis extract for two weeks did not result in macroscopic lesions of the gastric mucosa.     

Prevention of experimentally induced irritant contact dermatitis by extracts of Isatis tinctoria compared to pure tryptanthrin and its impact on UVB-induced erythema

Lipophilic extracts of Isatis tinctoria L. exhibit significant activity against several clinically relevant targets of inflammation. The alkaloid tryptanthrin was identified as one of the active principles in woad and characterised as a potent dual inhibitor of COX-2 and 5-LOX. Here, the anti-inflammatory efficacy of topical application of three different Isatis extracts and tryptanthrin was investigated in human volunteers. Two different models were used, namely the sodium lauryl sulphate (SLS)-induced irritant contact dermatitis (ICD) and UVB-induced erythema. Twenty healthy volunteers without any skin disease participated in the study. Cumulative irritant contact dermatitis was induced on test fields on the volunteers' backs by twice daily application of 0.5 % sodium lauryl sulphate over a period of four days. Half of the test fields were treated with the test substances during the eliciting phase, while the remaining test fields were treated over a period of 4 days after induction of dermatitis. In the second model, a UVB erythema on the volunteers' lower backs was induced using the double minimal erythema dose (MED). Twenty-four hours after irradiation the test fields were treated with the test substances over a period of 3 days. All reactions were assessed visually and by non-invasive bioengineering methods (evaporimetry and chromametry). Treatment with extracts during the ICD eliciting phase led to a significantly smaller increase of visual scores and transepidermal water loss compared to the untreated test field. For tryptanthrin this benefit was also observed, but the improvement was not statistically significant. When treatment was performed after completing the eliciting phase, accelerated resolution of the irritant reaction could not be observed. In the UVB erythema model anti-inflammatory effects of the test substances were not observed.     

A comparative study on the skin penetration of pure tryptanthrin and tryptanthrin in Isatis tinctoria extract by dermal microdialysis coupled with isotope dilution ESI-LC-MS

The indolo[2,1- b]quinazoline alkaloid tryptanthrin has recently been identified as a pharmacologically active compound in Isatis tinctoria, with potent dual inhibitory activity on prostaglandin and leukotriene synthesis. To investigate the skin penetration of tryptanthrin from solutions of pure compound and Isatis extracts, we developed and validated a cutaneous microdialysis model using ex vivo pig foreleg. Microdialysis was performed by placing linear probes in the dermis of the skin in situ, and tryptanthrin concentrations in the dialysates were determined by isotope dilution electrospray ionization LC-MS in the selected ion mode. Measurable concentrations of tryptanthrin were detected 30 min after application. A dose-dependent increase in tryptanthrin concentrations in the dialysate was observed for the Isatis extracts, but not for pure tryptanthrin. Microscopic analysis showed that the pure compound crystallized from the solution but remained in an amorphous state in the extracts.     

Quantitative determination of the dual COX-2/5-LOX inhibitor tryptanthrin in Isatis tinctoria by ESI-LC-MS

Isatis tinctoria L. is an old European and Chinese dye plant and anti-inflammatory herb from which the potent cyclooxygenase-2 and 5-lipoxygenase inhibitor tryptanthrin (1) (indolo-[2,1-b]-quinazoline-6,12-dione) was recently isolated as one of the active principles. An HPLC method for the quantitative analysis of the compound in plant material was developed. Reproducible extraction was achieved by accelerated solvent extraction (ASE). Detection by UV at 254 and 387 nm and by electrospray-MS were compared. The low tryptanthrin content in the herb and possible interferences required isocratic high-performance liquid chromatography coupled with electrospray-MS in single ion mode. More than 70 Isatis samples of different origin were analyzed. The tryptanthrin content in leaf samples varied from 0.56 to 16.74 x 10(-3) %.     

Quantification of active principles and pigments in leaf extracts of Isatis tinctoria by HPLC/UV/MS

An HPLC method has been developed and validated for the quantification of the pharmacologically active principles tryptanthrin (1), 1,3-dihydro-3-[(4-hydroxy-3,5-dimethoxyphenyl)methylene]-2 H-indol-2-one (indolinone) (3), indirubin (4), alpha-linolenic acid (2), and indigo (5), an isomer of indirubin, in extracts from the traditional anti-inflammatory plant Isatis tinctoria (woad). The chromatographic separation was performed on a C-18 column with a linear gradient of acetonitrile in water containing 0.1% formic acid. The method combines UV and electrospray MS detection in the positive ion mode for the detection of the alkaloids, with a switch to the negative mode for the analysis of alpha-linolenic acid. The method was applied to the analysis of woad extracts obtained by supercritical fluid (SFE) CO2 extraction, and by pressurized liquid extraction (PLE) with dichloromethane and methanol, respectively. While the highest concentration of alpha-linolenic acid was found in the SFE extract (7.43%), the concentrations of tryptanthrin , indolinone, indirubin and indigo were the highest in the dichloromethane extract (0.30, 0.035, 2.48 and 0.84%, respectively). Compound 3 was not detected in the methanolic extract and only traces of compounds 1, 4 and 5 and low amount of alpha-linolenic acid (0.39%) were present in this extract.     

Glucosinolate pattern in Isatis tinctoria and I. indigotica seeds

The glucosinolate patterns in seeds of five ISATIS TINCTORIA and two ISATIS INDIGOTICA accessions were assessed with a recently developed and validated LC-MS assay for direct analysis of glucosinolates without prior desulfatation. Glucosinolate peaks were identified with in-source fragmentation and detection of the sulfate anion ( M/Z = 97), and by MS/MS experiments. The glucosinolate patterns of the seeds showed characteristic differences compared to leaves. Glucoisatisin and epiglucoisatisin were diagnostic of seed samples. Qualitative and quantitative differences in glucosinolate patterns between both ISATIS species were found for seed samples, enabling a differentiation of the two medicinal plants at the level of seed material.     

板蓝根饮片产地加工炮制技术研究

目的:筛选板蓝根饮片产地加工炮制技术,为保证板蓝根饮片质量提供依据。方法:对新鲜采收的板蓝根药材在4种不同干燥程度下切制后所得饮片[A.切片后60℃干燥;B.切片后自然干燥;C.自然干燥(13~15 d)后切片,再60℃干燥;D.自然干燥(30~50 d)后切片,再60℃干燥],从外观评分及水分、灰分、浸出物、主要成分(R,S)-告依春的含量方面进行比较。结果:各饮片外观性状评分从高到低为C>D>A>B,水分从高到低为B>A>D>C;灰分各法差别不大;浸出物和(R,S)-告依春含量从高到低均为C>A>B>D。结论:4种加工炮制技术所得板蓝根饮片质量不同,将鲜品自然干燥(约15 d)到一定程度后再切制及干燥后所得板蓝根饮片的质量相对最好。     

Anti-inflammatory activity of extracts from Conyza canadensis

The petroleum ether and ethanolic extract from the epigean part of Conyza canadensis exhibits a significant anti-inflammatory effect on rats with a carrageenin and formalin oedema. Eight sesquiterpenic hydrocarbons with the highest anti-inflammatory activity were found in the petroleum ether fraction (beta-santalene, beta-himachalene, cuparene, alpha-curcumene, gamma-cadinene and three other unidentified hydrocarbons). Of these substances, beta-himachalene was further studied and its anti-inflammatory activity was demonstrated.     

Identification and isolation of the cyclooxygenase-2 inhibitory principle in Isatis tinctoria

Various extracts prepared from the traditional dye and medicinal plant Isatis tinctoria L. were submitted to a broad in vitro screening against 16 anti-inflammatory targets. Dichloromethane (DCM) extracts from dried leaves showed a marked cyclooxygenase (COX) inhibitory activity with a preferential effect on COX-2 catalysed prostaglandin synthesis. A supercritical fluid extraction (SFE) procedure employing CO2-modifier mixtures was developed by which the bioactivity profile and chromatographic fingerprint of the DCM extract could be reproduced. High-resolution activity directed on-line identification of the COX-2 inhibitory principle, using a combination of LC-DAD-MS with a microtitre-based bioassay, led to the identification of tryptanthrin (1) as the constituent responsible for essentially all COX-2 inhibitory activity in the crude extract. Following on-line identification, 1 was isolated at preparative scale and its structure confirmed by comparison with synthetic tryptanthrin. In an assay with lipopolysaccharide stimulated Mono Mac 6 cells, tryptanthrin (1) was of comparable potency (IC50 = 64 nM) than the preferential COX-2 inhibitors nimesulide (IC50 = 39 nM) and NS 398 (IC50 = 2 nM). The SFE extract and 1 showed no cytotoxicity in Mono Mac 6 and RAW 264.7 cells when tested at 100 microg/ml and 10 microM, respectively.     

Anti-inflammatory activity of aqueous and ethanolic extracts of Zygophyllum gaetulum

Zygophylle or Zygophyllum gaetulum Emberger and. Maire is a Moroccan medicinal plant which has been used as an anti-inflammatory, antidiabetic, antispasmodic and antidiarrheic. The present study was carried out to study and compare the anti-inflammatory effect of ethanolic extract with aqueous extract of Z. gaetulum. Organic extract of Z. gaetulum was obtained in soxhlet apparatus. Aqueous extract was obtained by infusion. The Wistar albinos rats of either sex weighing 200-300 g aged 2-3 months were used for this experiment. The rats were housed under standard environmental conditions. The anti-inflammatory activity was estimated by measuring the oedema induced by carragenin according to the method of Winter and al. Ethanolic extract of Z. gaetulum reduced the increase of the paw volume with a percentage of inhibition of 46% (p<0.01), this percentage was 47.48% (p<0.01) with aqueous extract. The inhibition decrease in time, it arrived to 39% (p<0.01) at the sixth hour while the activity of aqueous extract decrease a lot. In conclusion, Z. gaetulum is an interesting plant which the aqueous and éthanolic extracts could be used scientifically in the treatment of inflammation.     

板蓝根不同提取部位抗菌活性的实验性研究

目的研究并比较板蓝根不同提取部位的抗菌作用。方法以不同极性溶剂革取板蓝根．得水煎液总提取物、石油醚提取物、乙酸乙酯提取物、氯仿提取物、正丁醇提取物及革取后物质等不同极性部位。以金黄色葡萄球菌和大肠埃希菌为试验对象,采用试管稀释法比较板蓝根不同提取部位的抗菌作用。结果板蓝根水提部位、乙酸乙酯部位、正丁醇部位、氯仿部位及石油醚部位均具有一定的抗菌作用．而革后部位没有抗菌作用。其活性比较：正丁醇部位〉乙酸乙酯部位〉氯仿部位〉水提部位〉石油醚部位。结论板蓝根能有效抑制金葡球菌和大肠埃希菌。具有广谱抗菌作用。板蓝根抗菌作用并不局限于一个部位,它是通过多种有效成分、多环节、多途径地发挥协同作用而表现出抗菌功效。     

Tryptanthrin content in Isatis tinctoria leaves--a comparative study of selected strains and post-harvest treatments

Tryptanthrin is a pharmacologically active compound in the anti-inflammatory herb Isatis tinctoria, with potent inhibitory activity on prostaglandin and leukotriene synthesis and on inducible NO synthase. The tryptanthrin content of five defined woad strains was analyzed in dependence of the time of harvest and post-harvest treatment. Tryptanthrin was determined by a validated ESI-LC-MS isotope dilution assay with d(8)-tryptanthrin as internal standard. The tryptanthrin concentration in freeze-dried leaf samples was low. Drying at ambient temperature led to a significant increase of tryptanthrin concentration, but the highest concentrations were found when leaves were dried at 40 degrees C. Tryptanthrin content in fermented woad leaves was below the limit of quantification. Tryptanthrin appears thus to be a product of post-harvest processes, but details of its formation remain to be elucidated.     

Anti-inflammatory activity of extracts and a saponin isolated from Melilotus elegans

The crude methanol extract of Melilotus elegans Ser. (Fabaceae), a plant widely used in Ethiopian traditional medicine for the treatment of asthma, haemorrhoid and lacerated wounds showed a significant anti-inflammatory activity against carrageenin-induced rat paw oedema. At a dose corresponding to 333.3 mg per kg body weight of dry plant material, the methanol extract displayed a strong inhibitory effect that was comparable to the inhibitory effect of 1 mg/kg of indomethacin in the same test system. Bioassay guided fractionation of the alcoholic extract led to the isolation of an oleanene-type triterpene saponin identified as azukisaponin V (1) ((3-O-[alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D-glucopyranosyl(1 --> 2)-beta-D-glucuronopyranosyl]-soyasapogenol B). The structure of the compound was identified by using MS and extensive one- and two-dimensional NMR experiments (1H, 13C, COSY, HMQC, HMBC and NOESY). One hour after injection of carrageenin, inhibition of oedema exerted by 1 was approximately ten times higher than that of indomethacin on a molar basis.     

In vivo antimalarial activity of ginseng extracts

Context: Novel antimalarial agents are in demand due to the emergence of multidrug resistant strains. Ginseng, a medicinal plant with antiparasitic activity, contains components that can be used to treat the tropical disease malaria.

Objective: Ginsenosides and polysaccharides are active components of ginseng. This study aimed to elucidate the ability of these compounds to inhibit the replication of Plasmodium yoelii in an attempt to determine whether the medicinal uses of ginseng are supported by pharmacological effects. New antimalarial compounds may be developed from ginsenosides and water-soluble ginseng polysaccharides (WGP).

Materials and methods: Ginsenosides and ginseng polysaccharides were prepared from ginseng. Antimalarial activities were examined by 4-day tests and repository tests. Macrophage phagocytosis was tested in normal and malaria-bearing mice.

Results: Ginseng polysaccharides could inhibit residual malaria infection. After a 6-day treatment, the parasitemia reductions of WGP and acidic ginseng polysaccharide (WGPA) were 55.66% and 64.73% at 200?mg/kg/day, respectively. Ginsenosides showed significant antimalarial activity on early infection. Protopanaxadiol-type ginsenosides caused 70.97% chemosuppression at 50?mg/kg/day, higher than 52.8% of total ginsenosides at the same dose.

Discussion and conclusion: Protopanaxadiol-type ginsenosides have remarkably suppressive activity during early infection, while acidic ginseng polysaccharides have significant prophylactic activity against malaria by stimulating the immune system. We propose that the activity of ginsenosides is dependent upon non-specific carbohydrate interactions and that the activity of ginseng polysaccharides is due to immunological modulation. Ginsenosides and ginseng polysaccharides might have a potential application in antimalarial treatments.     

Antidyslipidemic activity of Indigofera tinctoria

Indigofera tinctoria is a perennial shrub, which belongs to the family Papilionaceae. As a part of our drug discovery program we have investigated the antidyslipidemic activity of the alcoholic extract from Indigofera tinctoria as well as its three other components, that is, chloroform, butanol and aqueous fractions in dyslipidemic hamsters that were fed a high fat diet. The chloroform fraction showed a significant decrease in the plasma triglycerides (TG, 52%) (P < 0.001), total cholesterol (TC, 29%) (P < 0.05), glycerol (Gly, 24%) and free fatty acids (FFA, 14%). This decrease was also accompanied by an increase in high density lipoproteins (HDL) by 9% and an increased HDL-C/TC ratio of 52% at the dose of 250 mg/kg of body weight.     

Inhibitory activity of tryptanthrin on prostaglandin and leukotriene synthesis

The indolo[2,1- b]quinazoline alkaloid tryptanthrin has previously been identified as the cyclooxygenase-2 (COX-2) inhibitory principle in the extract ZE550 prepared from the medicinal plant Isatis tinctoria (Brassicaceae). We here investigated the potential inhibitory activity of tryptanthrin and ZE550 on COX-2, COX-1 in cellular and cell-free systems. A certain degree of selectivity towards COX-2 was observed when COX-1-dependent formation of thromboxane B(2) (TxB(2)) in HEL cells and COX-2-dependent formation of 6-ketoprostaglandin F(1alpha) (6-keto-PGF(1alpha)) in Mono Mac 6 and RAW 264.7 cells were compared. Preferential inhibition of COX-2 by two orders of magnitude was found in phorbol myristate acetate (PMA) activated bovine aortic coronary endothelial cells (BAECs). Assays with purified COX isoenzymes from sheep confirmed the high selectivity towards COX-2. The leukotriene B(4) (LTB(4)) release from calcium ionophore-stimulated human granulocytes (neutrophils) was used as a model to determine 5-lipoxygenase (5-LOX) activity. Tryptanthrin and the extract ZE550 inhibited LTB(4) release in a dose dependent manner and with a potency comparable to that of the clinically used 5-LOX inhibitor zileuton.     

Anti-inflammatory activity of dichloromethane extract of Heterotheca inuloides in vivo and in vitro

The dichloromethane extract from the dried flowers of Heterotheca inuloides Cass. was investigated on several pharmacological models of inflammation in vivo and in vitro. It showed anti-inflammatory activity on the croton oil-induced oedema test in mouse ear, at 1 mg/ear. The compound isolated from this extract, 7-hydroxy-3,4-dihydrocadalin, showed anti-inflammatory effect on the same experimental model (ED50 of 0.9 mumol/ear), as well as on COX-1 and COX-2 catalysed prostaglandin biosynthesis assays, with IC50 values of 22 microM and 526 microM, respectively. No effect was observed on carrageenan-induced oedema and on fMLP/PAF-induced exocytosis of human neutrophils. The COX-1 inhibitory effect showed by 7-hydroxy-3,4-dihydrocadalin might be related to the anti-inflammatory activity on the topical oedema induced by croton oil.