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Characterisation and immuno-stimulating activity of polysaccharides from Thai medicinal plants

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Water-soluble polysaccharides were isolated from the tubers of Butea superba Roxb. and Pueraria candollei Wall. Ex Benth. var. mirifica (Shaw et Suvat.) C. Niyomdham, the leaves of Centella asiatica (L.) Urb, Ocimum basilicum L., Psidium guajava and Andrographis paniculata (Burn. f.) Nees, the stems of Cymbopogon citratus (Stapf ExG), and the fruits of Psidium guajava and Scaphium scaphigerum. The immunological impacts of the polysaccharides on T-lymphocyte proliferation in vitro was investigated by flow cytometric (immunofluorescence) analysis using staphylococcal enterotoxin B (SEB) as a positive control. It was found that the polysaccharides enhanced T-lymphocyte proliferation, ranging from 4.5 to 27.0% at a concentration of 100 microg mL(-1), while the activity of SEB was 13.3%. The medicinal plants showing the highest immuno-stimulating activity were the tubers of Butea superba Roxb. The water-extracted tubers contained 60.0% (w/w) carbohydrates with 6.6% (w/w) uronic acid. The major constituent monosaccharides of the tubers were 28.2 mol% galactose, 10.5 mol% arabinose and 36.4 mol% glucose.

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The expression of surface antigen CD69 in immune response cells is typically associated with the early stage(s) of cell activation, with maximal expression levels within 4 h of appropriate antigenic or mitogenic stimulation, and maintenance of these high expression levels for 18-24 h. The expression profiles of CD69 in human peripheral blood mononuclear cells (PBMC) cultured with actinomycin D prior to mitogenic stimulation were evaluated by direct immunofluorescence using flow cytometry. Pretreatment of PBMC suspensions with low, non-toxic levels of actinomycin D stimulated CD3+ T-lymphocytes to express CD69 in a concentration-dependent manner. Furthermore, CD4+ T-lymphocytes were the primary cells responding in this fashion. Secondary mitogenic stimulation following antibiotic treatment potentiated cellular CD69 expression in these assays. CD69 expression was profoundly suppressed with in vitro actinomycin D concentrations >/=1-2 microg/ml, presumably by interference with cellular transcription/translation mechanisms. Parallel thymidine incorporation assays indicated that actinomycin D effectively inhibited thymidine uptake in a concentration-dependent manner, with complete inhibition at >/=0.1 microg/ml. The evaluation of cell cycling dynamics following antibiotic treatment, with and without secondary mitogen stimulation, indicated no substantial changes in DNA synthesis over controls. The diversity of these responses suggests that expression of CD69 may not solely reflect mitogenic activation status but may, under some conditions, result from induced cellular stress.
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Hot water polysaccharide extracts of Anacyclus pyrethrum (L.) Link. (family Compositae) Citrullus colocynthis (L.) Schrad. (family Cucurbitaceae) and Alpinia galanga (L.) Willd. (family Zingiberaceae) were tested for their immunostimulating activity in mice. The fractions from Anacyclus pyrethrum and Alpinia galanga showed a marked stimulating effect on the reticulo-endothelial system (RES) and increased the number of peritoneal exudate cells (PEC), and spleen cells of mice. In this case, the optimum doses were 50 and 25 mg/kg for the two fractions, respectively. On the other hand, the polysaccharide extracts of both Anacyclus pyrethrum and Alpinia galanga markedly enhanced the proliferation of the murine spleen cells in vitro using two tests (in vitro and in vivo effect). The results of the in vivo effect at a doses of 50 and 25 mg/kg, showed a stimulation index better than obtained with the in vitro effect at 50 and 25 microg/ml for Anacyclus pyrethrum and Alpinia galanga, respectively. While the extract of Citrullus colocynthis showed much weaker and variable immunostimulating activity.
Article
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.
Article
Aeginetia indica Roxbert (Dok Din Daeng, DDD), a parasitic plant that grows on bamboo, is extensively used in Thai traditional medicine to treat various diseases. There have been no published studies on the pharmacological, toxicological or immunological effects of DDD, indigenous to Thailand. The study reported here was focused on the immunological effects (T cells) of the whole plant extract using water (WDDD) or ethanol (EDDD) as a solvent. The extracts were administered to female B6C3F1 mice by gavage for WDDD (10-100%) and intraperitoneally (i.p.) for EDDD (0.25-250 mg/kg) for 28 days. Only mice administrated the highest dose of EDDD exhibited an increase in absolute spleen and liver weights. Three T cell functional assays, including anti-CD3 antibody-mediated T cell proliferation, the mixed leukocyte response (MLR) and the cytotoxic T lymphocyte (CTL) response, were employed to determine the effects of DDD extracts on splenic T cell activities. Exposure to WDDD enhanced the responses in all three assays with significant changes observed in the anti-CD3 and MLR assays. Exposure to EDDD also enhanced the responses in all three assays with significant changes observed in the MLR and CTL assays. Additionally, significant increases in the MLR and anti-CD3 responses were also observed when EDDD was used to treat cells in vitro. Finally, exposure to WDDD decreased both the percentage and absolute number of regulatory CD4(+)CD25(+) T cells in the spleen, which was consistent with a significant increase in interferon-gamma (IFN-gamma) production from Con A-stimulated splenocytes. Overall, this study demonstrated that the extracts from A. indica Roxbert had a T cell stimulatory activity.
Thai plant names dictionary. Bangkok: The Forest Herbarium Royal Forest Department
  • T Smitinand
Smitinand, T. (2001). Thai plant names dictionary. Bangkok: The Forest Herbarium Royal Forest Department.
Thai medicinal plants: Recommended for primary health care system
  • N R Frnsworth
  • N Bunyapraphaisara
Frnsworth, N.R., Bunyapraphaisara, N. (1992). Thai medicinal plants: Recommended for primary health care system. Bangkok: Medicinal Plants Information Center, Mahidol University.