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1.
J Ethnopharmacol ; 152(2): 340-8, 2014 Mar 14.
Artículo en Inglés | MEDLINE | ID: mdl-24480566

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Sutherlandia frutescens (syn. Lessertia frutescens) is an indigenous plant in Southern Africa and has been extensively studied from the ethnobotanical point of view. Amongst the various traditional uses, several illnesses involving the immune system have been reported. Due to some of the therapeutic effects observed, in relation to the traditional uses reported by the "khoi san" and "nama" people on cancer related illnesses, the plant has been given the local name kankerbos (cancerbush). Recently the plant has also been used amongst HIV/AIDS patients to stimulate the immune system. MATERIALS AND METHODS: Leaves of Sutherlandia frutescens were extracted sequentially with ethanol, 50% ethanol/water, and water at 50 and 100°C. The polysaccharides were extracted with water and fractionated by ion exchange chromatography and gel filtration to obtain enriched polysaccharide fractions. The bioactivities of the fractions were tested in the complement assay. Some of the fractions were treated with the enzyme pectinase, and the fragments thus produced were separated by gel filtration and their activities tested. Monosaccharide compositions and linkage analyses were determined for the relevant fractions. RESULTS: The leaves of Sutherlandia frutescens contain polysaccharides of the pectin type. Fractions from both the water extracts of 50 and 100°C were bioactive. Fractions chosen for further studies showed that the fragment with the highest M(W) after the pectinase treatment had a substantially higher biological effect than the parent molecules. Based on a comparison of the different fractions it was concluded that galactose-rich regions were important for the bioactivity, these being of the AGII and AGI type, with the latter probably being more important than the former. Fragments rich in xylose also gave higher activity than those without it. CONCLUSIONS: Our theory that the polysaccharides present in the leaves of Sutherlandia frutescens could be of importance as immunomodulating agents was confirmed. It was also shown that certain types of polysaccharides had a higher effect in the complement system than others. Thus both the water extracts obtained at 50 and 100°C contain interesting biologically active polysaccharides.


Asunto(s)
Proteínas del Sistema Complemento/efectos de los fármacos , Fabaceae/química , Extractos Vegetales/farmacología , Polisacáridos/farmacología , Cromatografía por Intercambio Iónico , Proteínas del Sistema Complemento/inmunología , Etanol/química , Humanos , Factores Inmunológicos/química , Factores Inmunológicos/aislamiento & purificación , Factores Inmunológicos/farmacología , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Hojas de la Planta , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Relación Estructura-Actividad , Temperatura , Agua/química
2.
Phytochemistry ; 93: 136-43, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23582214

RESUMEN

In Malian traditional medicine the roots of Cochlospermum tinctorium are used in the treatment of gastric ulcer, but extending harvesting is causing a growing concern of a dramatic reduction in the wild plant population. In the present study cultivation of C. tinctorium is evaluated, and structural components and bioactive properties of crude water extracts and isolated polysaccharide fractions from roots of wild and cultivated C. tinctorium are compared. The crude water extracts were shown to contain starch, pectin- and inulin-type polysaccharides, in addition to phenolic substances and protein, while the isolated acidic polysaccharide fractions contained mainly monosaccharides typical for pectins. The monosaccharide compositions of the polysaccharide fractions from roots of wild versus cultivated plants were comparable, albeit the yields in the cultivated roots were lower. Furthermore, the crude extracts and isolated polysaccharide fractions from wild and cultivated roots exhibited similar complement fixating activities, but were not able to activate macrophages. The crude extracts from cultivated roots were also shown to be moderate radical scavengers. The present study has shown that roots of cultivated C. tinctorium contain the same types of bioactive polysaccharides as the wild roots. However, in order to utilize roots of cultivated C. tinctorium in traditional medicine the cultivation method should be improved.


Asunto(s)
Bixaceae/química , Macrófagos/efectos de los fármacos , Extractos Vegetales/farmacología , Raíces de Plantas/química , Polisacáridos/farmacología , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Ratones , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Relación Estructura-Actividad , Agua/química
3.
J Ethnopharmacol ; 143(1): 221-7, 2012 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-22732727

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Aqueous extracts of bark and leaves of C. cordifolia are traditionally used in Mali (West Africa) in the treatment of wounds and gastric ailments like abdominal pain, gastritis and gastric ulcers. AIM OF THE STUDY: To evaluate and compare the anti-ulcer and immunological activities, as well as the toxicity of polysaccharide rich water extracts from the bark and leaves of C. cordifolia. MATERIALS AND METHODS: Gastric ulcers were induced in rats and the inhibition of ulcer formation was calculated based on lesion index. Immunological activities were measured by complement fixation and macrophage activation. Toxicity was tested on brine shrimps. The two extracts were characterised by GC, Yariv-precipitation and quantification of phenolic compounds. An ethnomedical survey on C. cordifolia was carried out in Siby (Mali, West-Africa) to generate more knowledge about the traditional use. RESULTS: Bark and leaf extracts from C. cordifolia significantly inhibited the formation of gastric lesions in rodents in a dose depending manner. CCbark50 showed a high complement fixation activity in vitro. No toxicity was found. The ethnomedical survey showed that C. cordifolia was mainly used for treating pain and wounds. CONCLUSIONS: Our results shows that the bark and the leaves comprise a dose dependant anti-ulcer activity in an experimental rat model (no statistical difference between the plant parts). Clinical studies should be performed to evaluate the effect of both bark and leaves of C. cordifolia as a remedy against gastric ulcer in human.


Asunto(s)
Antiulcerosos/uso terapéutico , Cola/química , Fenoles/uso terapéutico , Fitoterapia , Extractos Vegetales/uso terapéutico , Polisacáridos/uso terapéutico , Úlcera Gástrica/tratamiento farmacológico , Animales , Antiulcerosos/farmacología , Artemia , Activación de Complemento/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Medicinas Tradicionales Africanas , Dolor/tratamiento farmacológico , Fenoles/farmacología , Corteza de la Planta , Extractos Vegetales/farmacología , Hojas de la Planta , Polisacáridos/farmacología , Ratas , Ratas Sprague-Dawley , Úlcera Gástrica/inmunología , Úlcera Gástrica/patología , Heridas y Lesiones/tratamiento farmacológico
4.
Carbohydr Polym ; 89(1): 259-68, 2012 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-24750632

RESUMEN

The bark of Cola cordifolia used in Malian traditional medicine contains unusual types of polysaccharides with immunomodulating activities. We report for the first time on the structure of a polymer designated CC1P1 having the repeating structure [2→)[α-D-Gal(1→3)]α-L-Rha(1→4)α-d-GalA(1→] as determined by NMR and GC/MS. α-Linked Gal is unusual in pectins. The Mw of 135 kDa was determined by SEC-MALLS. CC1P2 (1400 kDa), another polymer, having the same backbone, but this was substituted with α-4-OMe-GlcA, α-2-OMe-Gal and α-Gal as terminal units. CC1P1 shows a high complement-fixing activity, IC50 being 2.2 times lower than the positive pectin control PMII (IC50 appr. 71 µg/mL) while IC50 of CC1P2 is 1.8 times lower. The simple structure of CC1P1 did not activate macrophages, while CC1P2 (100 µg/mL) showed the same potency as the positive controls PMII (100 µg/mL) and LPS (500 ng/mL). No cytotoxicity was detected.


Asunto(s)
Cola , Polisacáridos/química , Polisacáridos/farmacología , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Esterificación , L-Lactato Deshidrogenasa/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Malí , Medicinas Tradicionales Africanas , Ratones , Monosacáridos/análisis , Óxido Nítrico/metabolismo , Corteza de la Planta/química , Plantas Medicinales , Polisacáridos/aislamiento & purificación , Relación Estructura-Actividad , Árboles
5.
Glycobiology ; 20(12): 1654-64, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-20729344

RESUMEN

The leaves of the tree Opilia celtidifolia have a long tradition for being used in Mali and other West African countries against various ailments such as for wound healing. Previous studies on polysaccharides from these leaves showed the presence of pectic-like polymers with an effect on the human complement system as well as the ability to activate macrophages. The present study shows that bioactive arabinogalactans isolated by water of 50°C could be separated into two acidic fractions, Oc50A1 and Oc50A2. The former could, by gel filtration on Sephacryl S-400, be separated into two fractions, which were further purified on a Superdex 200 column to give the fractions Oc50A1.I.pur and Oc50A1.II.pur. These fractions were subjected to chemical and biological studies. The polysaccharides consisted mainly of heavily branched type II arabinogalactans and minor amounts of rhamnogalacturonan I regions. The isolated polymers had a high human complement-fixating ability, as well as the ability to stimulate rat macrophages and dendritic cells (DCs) and to induce B cell proliferation. These effects were especially pronounced for the higher molecular weight fraction of Oc50A1.I.pur. The fractions Oc50A1.I.pur and Oc501.II.pur stimulated secretion of pro-inflammatory cytokines from purified B cells or DCs. Collectively, these results indicate that the arabinogalactan type II polymers present in the leaves of O. celtidifolia may be used to develop medical devices for regulating inflammatory processes.


Asunto(s)
Galactanos/química , Galactanos/farmacología , Extractos Vegetales/química , Extractos Vegetales/farmacología , Hojas de la Planta/química , Árboles/química , Animales , Linfocitos B/citología , Linfocitos B/metabolismo , Proliferación Celular/efectos de los fármacos , Activación de Complemento/efectos de los fármacos , Pruebas de Fijación del Complemento/métodos , Proteínas del Sistema Complemento/química , Proteínas del Sistema Complemento/metabolismo , Células Dendríticas/citología , Células Dendríticas/metabolismo , Galactanos/aislamiento & purificación , Humanos , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Activación de Macrófagos/efectos de los fármacos , Macrófagos/citología , Macrófagos/metabolismo , Malí , Extractos Vegetales/aislamiento & purificación , Ratas , Cicatrización de Heridas/efectos de los fármacos
6.
Glycobiology ; 18(12): 1074-84, 2008 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-18809620

RESUMEN

The Malian medicinal plant Biophytum petersianum Klotzsch (Oxalidaceae) is used as a treatment against various types of illnesses related to the immune system, such as joint pains, inflammations, fever, malaria, and wounds. A pectic polysaccharide obtained from a hot water extract of the aerial parts of B. petersianum has previously been reported to consist of arabinogalactans types I and II (AG-I and AG-II), probably linked to a rhamnogalacturonan backbone. We describe here further structural characteristics of the main polysaccharide fraction (BP1002) and fractions obtained by enzymatic degradations using endo-alpha-d-(1-->4)-polygalacturonase (BP1002-I to IV). The results indicate that in addition to previously reported structures, rhamnogalacturan type II and xylogalacturonan areas appear to be present in the pectic polymer isolated from the plant. Atomic force microscopy confirmed the presence of branched structures, as well as a polydisperse nature. We further tested whether the BP1002 main fraction or the enzymatically degraded products could induce immunomodulating activity through stimulation of subsets of leukocytes. We found that macrophages and dendritic cells were activated by BP1002 fractions, while there was little response of T cells, B cells, and NK cells. The enzymatic treatment of the BP1002 main fraction gave important information on the structure-activity relations. It seems that the presence of rhamnogalacturonan type I is important for the bioactivity, as the bioactivity decreases with the decreased amounts of rhamnose, galactose, and arabinose. The demonstration of bioactivity by the plant extracts might indicate the mechanisms behind the traditional medical use of the plant.


Asunto(s)
Células Dendríticas/efectos de los fármacos , Macrófagos/efectos de los fármacos , Magnoliopsida/química , Pectinas/química , Pectinas/farmacología , Animales , Células Presentadoras de Antígenos/metabolismo , Células Dendríticas/metabolismo , Leucocitos/metabolismo , Linfocitos/metabolismo , Macrófagos/metabolismo , Microscopía de Fuerza Atómica , Modelos Moleculares , Peso Molecular , Pectinas/aislamiento & purificación , Extractos Vegetales/química , Extractos Vegetales/metabolismo , Poligalacturonasa/metabolismo , Ratas , Ratas Endogámicas , Relación Estructura-Actividad
7.
J Ethnopharmacol ; 115(3): 423-31, 2008 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-18053663

RESUMEN

AIM OF THE STUDY: The present study is aimed to determine the bioactivity and structure of polysaccharides present in the leaves from the Malian medicinal plant Opilia celtidifolia [Guill. & Perr. Endl. ex Walp (Opiliaceae)]. MATERIALS AND METHODS: The polysaccharides from the leaves of Opilia celtidifolia were isolated from water extracts of the leaves using gelfiltration and anion exchange chromatography giving the fractions Oc50A1 and Oc50A2. Monosaccharide composition was determined by gas chromatography of the derived TMS-derivatives of the methyl-glycosides. Linkages were determined of the partly methylated, partly acetylated alditol acetates obtained after a process including reduction, methylation, hydrolysis, reduction and acetylation followed by GC-MS. Effects on the complement system and the macrophages were determined using specific methods aimed for studying those activities. RESULTS: The polysaccharide fractions isolated from the leaves of Opilia celtidifolia has high complement fixing activity and induce nitrite oxide release from macrophages in a dose dependent manner. The fractions had an ICH50 of 0.5 and 0.9 microg/ml respectively in the complement fixing assay. They induced the release of 7.2 and 7.3 microM of nitrite oxide from macrophages respectively at a dose of 100 microg/ml. The monosaccharide composition in Oc50A1 and Oc50A2, analysed, showed the presence of arabinose (26.7 and 13.2%), galactose (31.5 and 28%) and galacturonic acid (5.3 and 7.8%) respectively. The Yariv test confirmed the presence of arabinogalactan type II in both fractions. Structural analyses did also show the presence of terminal and 1-4 linked galacturonic acid and terminal and 1-2 linked rhamnose. Endo-polygalacturonanase treatment was performed to isolate the heavily substituted parts of the polysaccharides. These parts contained the same monosaccharides in similar proportion, and showed stronger dose dependent complement fixing activity and also stimulated macrophages to release nitrite oxide. CONCLUSIONS: The leaves of Opilia celtidifola contains polysaccharides of pectic type that have both complement fixing and macrophage stimulating activity.


Asunto(s)
Macrófagos/efectos de los fármacos , Monosacáridos/farmacología , Plantas Medicinales/química , Polisacáridos/farmacología , Animales , Pruebas de Fijación del Complemento , Relación Dosis-Respuesta a Droga , Eritrocitos/efectos de los fármacos , Eritrocitos/metabolismo , Hemólisis/efectos de los fármacos , Macrófagos/metabolismo , Malí , Medicinas Tradicionales Africanas , Monosacáridos/administración & dosificación , Monosacáridos/aislamiento & purificación , Óxido Nítrico/metabolismo , Hojas de la Planta , Polisacáridos/administración & dosificación , Polisacáridos/aislamiento & purificación , Ovinos
8.
Glycobiology ; 17(12): 1299-310, 2007 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-17726087

RESUMEN

The aim of this paper was to further elucidate the structure and the immunomodulating properties of the pectic polymer GOA2, previously isolated from Glinus oppositifolius. Enzymatic treatment of GOA2 by endo-alpha-d-(1 --> 4)-polygalacturonase led to the isolation of three pectic subunits, GOA2-I, GOA2-II, and GOA2-III, in addition to oligogalacturonides. GOA2-I was shown to consist of 1,2-linked Rhap and 1,4-linked GalpA in an approximately 1:1 ratio, and NMR-analysis showed that the monomers were linked together in a strictly alternating manner. The galactose units in GOA2-I were found as terminal-, 1,3-, 1,6-, 1,4-, 1,3,4-, and 1,3,6-linked residues, while the arabinofuranosyl existed mainly as terminal- and 1,5-linked units. A rhamnogalacturonan-I type structure was suggested being the predominant part of GOA2-I. According to linkage analysis GOA2-II and GOA2-III contained glycosidic linkages characteristic for rhamnogalacturonan-II type structures. GOA2 was shown by sedimentation velocity in the analytical ultracentrifuge, to have a broad degree of polydispersity with a mode s(20,w) value of approximately 1.9 S, results reinforced by atomic force microscopy measurements. The polydispersity, as manifested by the proportion of material with s(20,w) > 3 S, decreased significantly with enzyme treatment. The abilities of GOA2, GOA2-I, GOA2-II, and GOA2-III to induce the proliferation of B cells, and to exhibit complement fixing activities were tested. In both test systems, GOA2-I showed significantly greater effects compared to its native pectin GOA2. GOA2-I was in addition shown to exhibit a more potent intestinal immune stimulating activity compared to GOA2. The ability of GOA2 to induce secretion of proinflammatory cytokines was examined. Marked upregulations in mRNA for IL-1beta from rat macrophages and IFN-gamma from NK cells were found.


Asunto(s)
Molluginaceae/metabolismo , Pectinas/química , Polímeros/química , Animales , Proliferación Celular , Galactosa/química , Humanos , Inflamación , Células Asesinas Naturales/metabolismo , Macrófagos/efectos de los fármacos , Ratones , Monosacáridos/química , Extractos Vegetales/farmacología , Ratas
9.
Phytochemistry ; 68(7): 1046-58, 2007 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-17337024

RESUMEN

An immunomodulating pectic polymer, GOA1, obtained from the aerial parts of the Malian medicinal plant Glinus oppositifolius (L.) Aug. DC. (Aizoaceae) has previously been reported to consist of arabinogalactans type I and II, probably linked to a rhamnogalacturonan backbone. To further elucidate the structure of the polymer GOA1, enzymatic degradation studies and weak acid hydrolysis were performed. Five different glycosidases were used, endo-alpha-D-(1-->4)-polygalacturonase, exo-alpha-L-arabinofuranosidase, endo-alpha-L-(1-->5)-arabinanase, endo-beta-D-(1-->4)-galactanase and exo-beta-D-galactosidase. It appears that GOA1 may contain a structural moiety consisting of a 1,3-linked galactopyranosyl (Galp) main chain with 1,6-linked Galp side chains attached to position 6 of the main chain. The 1,6-linked Galp side chain may be branched in position 3 with arabinofuranosyl (Araf) side chains. A 1,4-linked Galp backbone which might carry side chains or glycosyl units attached to position 3 is also a structural element in the polymer. We further show that GOA1 induce proliferation of B cells and the secretion of IL-1beta by macrophages, in addition to a marked increase of mRNA for IFN-gamma in NK-cells. To elucidate structure-activity relations the native polymer and the digested fractions were tested for complement fixing activity and intestinal immune stimulating activity. The partial removal of Araf residues after enzymatic degradations did not affect the bioactivities, while the acid hydrolysed fraction showed reduced complement fixing activity. A decrease in Araf units, 1,3,6-linked Galp units and a partial hydrolysed rhamnogalacturonan backbone, in addition to a reduction in molecular weight are factors that might have contributed to reduced bioactivity.


Asunto(s)
Aizoaceae/química , Factores Inmunológicos/química , Pectinas/química , Animales , Línea Celular , Línea Celular Tumoral , Citocinas/genética , Relación Dosis-Respuesta a Droga , Femenino , Galactanos/química , Galactanos/farmacología , Expresión Génica/efectos de los fármacos , Humanos , Hidrólisis , Factores Inmunológicos/farmacología , Células Asesinas Naturales/efectos de los fármacos , Células Asesinas Naturales/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Espectroscopía de Resonancia Magnética , Ratones , Ratones Endogámicos C3H , Pectinas/farmacología , Ganglios Linfáticos Agregados/efectos de los fármacos , Ganglios Linfáticos Agregados/inmunología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
10.
Biomacromolecules ; 8(2): 644-9, 2007 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-17253763

RESUMEN

Leaves of different cabbage species are used both as food and as wound healing remedies in traditional medicine. This supposed wound healing activity might be connected to presence of immunomodulating water soluble polysaccharides. To study this, three different cabbage varieties, white cabbage (W), kale (K), and red kale (RK), were pretreated with 80% ethanol and then extracted with water at 50 degrees C and 100 degrees C for isolation of polysaccharide-containing fractions. The fractions were analyzed for monosaccharide composition, glycosidic linkages, Mw distribution, protein content, and phenolic compounds and then tested for complement-fixing activity. All fractions contained pectin type polysaccharides with linkages corresponding to homogalacturonan and hairy regions. Those extracted at 50 degrees C contained higher amounts of neutral side chains and were more active in the complement-fixation test than those extracted at 100 degrees C. The fractions can be ranged by decreasing activity: K-50 > RK-50 > W-50 approximately = K-100 > RK100 approximately = W-100. Studies on structure-activity relationships (SAR) employing multivariate statistical analysis strongly suggest that the magnitude of the measured activity is influenced by the content of certain side chains in the polymers. High activity correlates to large neutral side chains with high amounts of (1-->6)- and (1-->3,6)-linked Gal and low amounts of (1-->4)-linked GalA but not on molecular weight distribution of the polymers.


Asunto(s)
Brassica/química , Proteínas del Sistema Complemento/metabolismo , Pectinas/química , Pectinas/inmunología , Pruebas de Fijación del Complemento , Galactosa , Cicatrización de Heridas
11.
Mol Nutr Food Res ; 50(8): 746-55, 2006 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-16865748

RESUMEN

This study was done to investigate whether white cabbage contained polysaccharides with immunostimulatory activity using the complement-fixing test as an indicator. The main polysaccharide isolated was of pectin nature. Methanolysis and (13)C-NMR showed that the polymers consisted of highly esterified alpha-galactopyranoside (alpha-GalpA), significant amounts of alpha-arabinose furanoside (alpha-Araf), beta-Galp and lesser amounts of rhamnose in the pyranose form (Rhap) and xylose in the pyranose form (Xylp). Linkage analyses showed that the alpha-GalpA residues were mainly 1,4-linked with small amounts of 1,3,4-linkages. The alpha-Araf residues were mainly terminally (t)- and 1,5-linked, whereas beta-Galp was t-, 1,3-, 1,6-, and 1,3,6-linked. Positive Yariv reaction indicated polymers with arabinogalactan type 2 like structures. alpha-Rhap was mainly present as 1,2- and 1,2,4-linked residues and Xylp was t- and 1,4-linked. The molecular weight varied greatly and was from 10 to 150 kDa. Cabbage polymers had biological activity and this complement-fixing activity was greatly affected by hydrolytic removal of Araf from pectic side chains.


Asunto(s)
Brassica/química , Proteínas del Sistema Complemento/efectos de los fármacos , Pectinas/química , Pectinas/aislamiento & purificación , Arabinosa/química , Conformación de Carbohidratos , Galactosa/química , Hidrólisis , Espectroscopía de Resonancia Magnética , Pectinas/farmacología , Polímeros/química , Ramnosa/química , Almidón/análisis
12.
Biomacromolecules ; 7(1): 48-53, 2006 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-16398497

RESUMEN

Biophytum petersianum Klotzsch (syn. Biophytum sensitivum (L.) DC) is a medicinal plant having a traditional use, among others, as a wound healing remedy in Mali and other countries. As a water extract of the aerial parts of the plant is a frequently used preparation, we decided to look for a bioactive polysaccharide in this extract. One of the obtained polysaccharide fractions, BP100 III, isolated from a 100 degrees C water extract from the aerial parts of B. petersianum and having a monosaccharide composition typical for pectic substances, was shown to exhibit potent dose-dependent complement fixating activity. The BP100 III fraction was subjected to degradation by endo-alpha-d-(1-->4)-polygalacturonase, and three fractions were obtained by gel filtration. The highest molecular weight fraction, BP100 III.1, had a more potent activity in the complement test system than the native polymer, while the two lower molecular weight fractions were less active than the native polymer. The major part of BP100 III.1 consists of galacturonic acid and rhamnose, with branches being present on both the rhamnose and galacturonic acid residues. Arabinogalactan type II is also present in the polymer, indicating that BP100 III.1 has a structure typical of the hairy region of pectins. The major part of the two other fractions is a galacturonan, containing a strikingly high number of branch points, some to which xylose is attached. These results indicate that the pectic substance in B. petersianum contains both rhamnogalacturonan and xylogalacturonan regions.


Asunto(s)
Proteínas del Sistema Complemento/inmunología , Magnoliopsida/química , Plantas Medicinales/química , Polisacáridos/química , Polisacáridos/inmunología , Animales , Pruebas de Fijación del Complemento , Humanos , Hidrólisis , Espectroscopía de Resonancia Magnética , Malí , Peso Molecular , Monosacáridos/química , Poligalacturonasa/metabolismo , Polisacáridos/aislamiento & purificación , Polisacáridos/metabolismo , Ovinos
13.
Biomacromolecules ; 7(1): 71-9, 2006 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-16398500

RESUMEN

Structures of three pectic arabinogalactans, one from Vernonia kotschyana (Vk2a) and two from Cochlospermum tinctorium (Ct50A1 and Ct50A2), and their complement fixation and induction of B cell proliferation in vitro were compared. The polysaccharide Vk2a expressed potent biological activity in both assays compared with Ct50A1 and Ct50A2. Vk2a possessed a very high molecular weight (1150 +/- 20 kDa) compared with Ct50A1 and Ct50A2 which both showed a polydisperse nature with the highest molecular weight polymers in each fraction estimated at approximately 105 kDa (Ct1a) and 640 +/- 100 kDa (Ct2a), respectively. The HMW polymers showed complement fixation in the same range as the native fractions. The arabinogalactan II content was low in Vk2a (2%) compared with that in Ct50A1 (23%) and Ct50A2 (12%). The high molecular weight polymers were subjected to digestion with a beta-d-(1, 3)-galactanase-rich fraction from Driselase, oligomers were isolated by HPAEC, and their finer structures were determined by MALDI- and ES-qoToF-MS, linkage, and monosaccharide composition analyses. Vk2a consists of both a galacturonan core and a rhamnogalacturonan core rich in neutral side chains. The backbones of both Ct-polysaccharides consist mainly of RG-I regions with numerous neutral side chains dominated by galactosyl residues, whereas the homogalacturonan regions seem to be small. Differences in the chain lengths of the 6-linked galacto-oligosaccharides attached to the 3-linked galactan core could not be related to the differences in the potencies of the biological activities observed.


Asunto(s)
Bixaceae/química , Proteínas del Sistema Complemento/efectos de los fármacos , Proteínas del Sistema Complemento/inmunología , Galactanos/química , Galactanos/farmacología , Mitógenos/farmacología , Vernonia/química , Aminoácidos/química , Animales , Antiulcerosos/química , Antiulcerosos/farmacología , Linfocitos B/citología , Linfocitos B/efectos de los fármacos , Linfocitos B/inmunología , Células Cultivadas , Pruebas de Fijación del Complemento , Femenino , Glicosilación , Espectroscopía de Resonancia Magnética , Ratones , Mitógenos/química , Mitosis/efectos de los fármacos , Peso Molecular , Monosacáridos/química , Océanos y Mares , Pectinas/química , Pectinas/farmacología , Relación Estructura-Actividad
14.
J Ethnopharmacol ; 101(1-3): 204-14, 2005 Oct 03.
Artículo en Inglés | MEDLINE | ID: mdl-15996844

RESUMEN

Glinus oppositifolius (L.) Aug. DC. (Aizoaceae) is a Malian medicinal plant used against various types of illnesses related to the immune response, like joint pains, inflammations, fever, malaria and wounds. Two pectin type polysaccharides, GOA1 and GOA2, being isolated from a 50 degrees C water extract from the aerial parts of Glinus oppositifolius were investigated for their activity towards the complement system and different leukocyte subsets because of the assumed effects on conditions related to the immune system. The polysaccharide polymer in GOA1 was shown to contain considerable amounts of the neutral sugars arabinose (26.4 mol%) and galactose (42.9 mol%), and methylation analysis indicated the presence of arabinogalactans type I (AG-I) and type II (AG-II). GOA2 was rich in galacturonic acid (68.3 mol%), along with rhamnose, arabinose and galactose. Structural studies indicated that rhamnose and galacturonic acid might constitute a rhamnogalacturonan backbone, often found in pectic substances, with side chains consisting of arabinose and galactose. Both GOA1 and GOA2 were shown to exhibit potent dose-dependent complement fixating activities, and induced chemotaxis of macrophages, T cells and NK cells.


Asunto(s)
Molluginaceae/química , Pectinas/aislamiento & purificación , Aminoácidos/análisis , Carbohidratos/análisis , Quimiotaxis/efectos de los fármacos , Pruebas de Fijación del Complemento , Humanos , Peso Molecular , Óxido Nítrico/análisis , Pectinas/análisis , Pectinas/química , Pectinas/farmacología , Proteínas/análisis
15.
Carbohydr Res ; 340(11): 1789-801, 2005 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-15979597

RESUMEN

Structure and immunological characteristics of the pectic arabinogalactan Vk2a (previously reported as Vk100A2a) from the roots of Vernonia kotschyana Sch. Bip. ex Walp. were investigated after enzymatic digestion of the galacturonan moiety and the side chains of the rhamnogalacturonan structure of Vk2a. endo-alpha-D-(1-->4)-Polygalacturonase digestion released the high molecular weight 'hairy region' (Vk2a-HR) and oligogalacturonides. Vk2a-HR consisted of GalA (4-linked) and Rha (2- or 2,4-linked) in a 1:1 ratio, with 60% of Rha branched at C-4. The Rha located in the rhamnogalacturonan core was branched randomly by Gal units. Vk2a-HR was rich in neutral sugars such as Araf 5- (12.2%) and 3,5-substituted (12.8%) and terminally- (14.1%) linked and Gal 4- (13.0%), 3- (0.9%), 6- (2.2%) and 3,6- (1.1%) substituted. Arabinans with chain lengths up to 11 units were identified. Araf residues were attached to C-3 of alpha-L-(1-->5)-Araf chains and to C-4 of Gal residues. Single Gal units and chains of beta-D-(1-->6)-linked galacto di- to penta-saccharides were attached to a beta-D-(1-->3)-galactan core. All the enzyme resistant fractions expressed potent complement fixation and induction of B-cell mitogenic activity, and the present study indicates that there may be several and possibly structurally different active sites involved in the bioactivity of Vk2a. The bioactive sites may be located both in the more peripheral parts of the molecule but also in the inner core of the 'hairy region' or in larger enzyme-resistant chains.


Asunto(s)
Galactanos/química , Pectinas/química , Animales , Sitios de Unión , Conformación de Carbohidratos , Carbohidratos/química , Cromatografía en Gel , Cromatografía Líquida de Alta Presión , Proteínas del Sistema Complemento/química , Eritrocitos/metabolismo , Galactosa/química , Cromatografía de Gases y Espectrometría de Masas , Humanos , Iones , Espectrometría de Masas , Metilación , Ratones , Ratones Endogámicos C3H , Mitógenos , Oligosacáridos/química , Raíces de Plantas/metabolismo , Polisacáridos/química , Ovinos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Relación Estructura-Actividad
16.
Carbohydr Res ; 340(1): 115-30, 2005 Jan 17.
Artículo en Inglés | MEDLINE | ID: mdl-15620674

RESUMEN

Two polysaccharides, a pectin (Vk100A2b) and a pectic arabinogalactan (Vk100A2a) with mean Mw 2 x 10(4) and 1.15 x 10(6)Da, respectively, were isolated from the dried powdered roots of Vernonia kotschyana Sch. Bip. ex Walp. by hot water extraction followed by fractionation on DEAE-Sepharose fast flow and Sephacryl S-400 HR. The pectin showed low-complement fixation activity and no influence on proliferation of B or T cells, while the pectic arabinogalactan showed a potent, dose-dependent complement fixation activity and a T cell independent induction of B-cell proliferation. Both polysaccharides induced chemotaxis of human macrophages, T cells and NK cells. exo-alpha-L-arabinofuranosidase and exo-beta-D-galactosidase digestion followed by component sugar and methylation analysis indicated that Vk100A2a consisted of a highly branched rhamnogalacturonan core with approximately 50% of the rhamnose 1,2,4-substituted, side chains rich in terminal-, 1,5-linked and 1,3,5-branched arabinose and terminal-, 1,4-, 1,6-linked and 1,3,6-branched galactose. The enzyme resistant part of Vk100A2a still showed strong complement fixating activity, suggesting that this activity may at least in part be expressed by carbohydrate structures present in the enzyme resistant, inner portion of the polymer.


Asunto(s)
Galactanos/química , Galactanos/inmunología , Inmunidad/inmunología , Pectinas/química , Pectinas/inmunología , Vernonia/química , Vernonia/clasificación , Quimiotaxis , Pruebas de Fijación del Complemento , Proteínas del Sistema Complemento/inmunología , Citometría de Flujo , Galactanos/aislamiento & purificación , Galactanos/metabolismo , Humanos , Macrófagos/citología , Macrófagos/inmunología , Óxido Nítrico/análisis , Pectinas/aislamiento & purificación , Pectinas/metabolismo , Raíces de Plantas , Linfocitos T/citología , Linfocitos T/inmunología
17.
Carbohydr Res ; 339(4): 753-62, 2004 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-14980816

RESUMEN

Crude water-soluble polysaccharides were isolated from Acanthus ebracteatus by hot water extraction followed by ethanol precipitation after pre-treatment with 80% ethanol. The crude polysaccharides were separated into neutral and acidic polysaccharides by anion-exchange chromatography. The neutral polysaccharide (A1001) was rich in galactose, 3-O-methylgalactose and arabinose, whereas the acidic polysaccharide (A1002) consisted mainly of galacturonic acid along with rhamnose, arabinose and galactose as minor components indicating a pectin-type polysaccharide with rhamnogalacturonan type I (RG-1) backbone. 3-O-Methylgalactose is also present in the acidic fraction. Both neutral and acidic fractions showed potent effects on the complement system using pectic polysaccharide PM II from Plantago major as a positive control. A small amount of 3-O-methylgalactose present in the pectin seemed to be of importance for activity enhancement in addition to the amount of neutral sugar side chains attached to RG-1. The relationship between chemical structure and effect on the complement system of the isolated polysaccharides is considered in the light of these data. The presence of the rare monosaccharide 3-O-methylgalactose may indicate that this can be used as a chemotaxonomic marker. The traditional way of using this plant as a medical remedy appears to have a scientific basis.


Asunto(s)
Acanthaceae/química , Tallos de la Planta/química , Plantas Medicinales/química , Polisacáridos/química , Polisacáridos/farmacología , Cromatografía en Gel , Cromatografía por Intercambio Iónico , Pruebas de Fijación del Complemento , Proteínas del Sistema Complemento/efectos de los fármacos , Cromatografía de Gases y Espectrometría de Masas , Hemólisis/efectos de los fármacos , Concentración de Iones de Hidrógeno , Espectroscopía de Resonancia Magnética , Peso Molecular , Polisacáridos/aislamiento & purificación , Refractometría
18.
J Ethnopharmacol ; 84(2-3): 279-87, 2003 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-12648827

RESUMEN

Trichilia emetica is a tree or bush that has many different traditional uses in Mali, amongst others the leaves have been used as a woundhealing remedy. As polysaccharides have been isolated from other plants that have a long tradition as woundhealing remedies, it was of interest to study the polysaccharides of T. emetica as well. The polysaccharides were extracted at 50 and 100 degrees C, and the polymeric material was separated into neutral and acidic polymers by anion exchange chromatography. The complement fixation ability of the different fractions was determined. The Acidic fraction 4 of the 100 degrees C extract was the most active, but most of the other fractions were also quite active. Structural studies showed that the most active fraction was a pectin of the rhamnogalacturonan type 1 with side chains of the arabinogalactan type II. Removal of terminal arabinofuranosides lead to a drop of the activity indicating that this structural unit may be involved in the bioactive site of the molecule.


Asunto(s)
Pruebas de Fijación del Complemento , Meliaceae/química , Polisacáridos/farmacología , Animales , Proteínas del Sistema Complemento/efectos de los fármacos , Eritrocitos/inmunología , Humanos , Hidrólisis , Malí , Peso Molecular , Monosacáridos/química , Hojas de la Planta/química , Polisacáridos/química , Ovinos/inmunología
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