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Objective: To investigate the immunomodulatory activity of polysaccharides from the roots of Brassica rapa. Methods: The crude polysaccharide from roots of B. rapa (BRP) was extracted and purified to further investigate the active fraction of BRT for inducing macrophage phagocytosis. Results: Effects on RAW264.7 cells demonstrated that BRP behaved better phagocytic capacity and had potent immunomodulatory activity, including increasing production of nitric oxide (NO), tumor necrosis factor α (TNFα) and upregulating mRNA levels of inducible NO synthase (iNOS) and TNFα. Furthermore, modulation of macrophage by BRP was indicated to be mediated via the activation of Akt and nuclear factor-kappa B (NF-κB). Conclusion: The beneficial effects of BRP could be used as an immunotherapeutic adjuvant in treatment of inflammatory diseases.
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Abstract The use of Echinacea purpurea (EP), a plant native from North America, is widely diffused throughout the world, presenting many pharmacological applications, mainly for the treatment of infections of respiratory and urinary tracts. Due to the widespread commercialization of EP-based products, an effective evaluation of their pharmacological properties is essential to assure efficacy during clinical use. In this study, in vitro tests were performed to evaluate the antimicrobial activity of dried extracts of EP by the microdilution method. In addition, a phagocytosis model was employed to assess the immunomodulatory potential of the extracts. The increase in reactive oxygen species production, as well as the intracellular proliferation rate of Cryptococcus gatti after phagocytosis by macrophages in the presence of EP dried extracts were also evaluated. The analyzed samples showed no significant antibacterial activity; however, a slight antifungal activity was verified. Positive effects of EP extracts on the modulation of cellular immune response were observed in different experiments, indicating that this mechanism may contribute to the control and treatment of infections.
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OBJECTIVE@#To evaluate the cytotoxic, apoptotic, mutagenic and immunomodulatory activities of Kaempferia galanga Linn. (KG) extract and ethyl-p-methoxycinnamate (EPMC) in vitro.@*METHODS@#The present study investigated the cytotoxic [using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide test], apoptotic (using a mitochondrial membrane potential assay), mutagenic (using a micronucleus test) and immunomodulatory (using flow cytometry) activities of the ethanolic extract of KG and its bioactive component, EPMC, against two cholangiocarcinoma (CCA) cell lines, CL-6 and HuCCT1, and one normal human cell line, OUMS-36T-1F.@*RESULTS@#Both KG extract and EPMC exhibited moderate cytotoxic activity against both CCA cells. The cytotoxic activity was supported by their concentration-dependent induction of apoptosis. CL-6 was most sensitive (3-4 fold) and selective to 5-fluorouracil (5-FU), compared with KG extract and EPMC [median half inhibiting concentration (IC) and selectivity index (SI) were 23.01 μg/mL and 17.32; 78.41 μg/mL and 4.44; 100.76 μg/mL and 2.20, respectively for 5-FU vs. KG extract vs. EPMC]. HuCCT1 was relatively more sensitive and selective to 5-FU and EPMC than KG extract [median IC and SI were 66.03 μg/mL and 6.04; 60.90 μg/mL and 3.65; 156.60 μg/mL and 2.23, respectively for 5-FU vs. EPMC vs. KG extract]. EPMC produced relatively potent cytotoxic activity against polymorphonuclear cells (IC = 92.20 μg/mL). KG extract and EPMC exhibited concentration-dependent mutagenic activity, as well as inhibition of tumor necrosis factor-α and interleukin-6.@*CONCLUSION@#Considering cytotoxic, apoptotic, immunomodulatory and mutagenic activities, further development of KG as a drug candidate is likely to focus on the oral pharmaceutical formulation of a standardized KG extract rather than isolated compounds.
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Objective: To extract and separate a polysaccharide from Polygonum multiflorum, characterize its structural features and study its immunomodulatory activity. Methods The polysaccharide from P. multiflorum (PMT) was isolated and purified by water extraction and ethanol precipitation following Q-Sepharose Fast Flow ion exchange chromatography column. Molecular weight of PMT was determined by high performance gel permeation chromatography-multiple angle laser light scattering (HPGPC-MALLS), and monosaccharide composition was analyzed by HPLC with PMP (1-phenyl-3-methyl-5-pyrazolone) pre-column derivatization, respectively. The structure of PMT was characterized by proton nuclear magnetic resonance spectrum (2D-NMR). The immunomodulatory activities were tested by MTT, neutral red colorimetric assay and Griess method. Results: PMT was a kind of α-1,4-glucan, and its molecular mass was 3.96 × 105. PMT promoted the proliferation and phagocytosis of RAW 264.7 cells, and significantly induced the increase of NO production in a dose-dependent manner. Conclusion: The polysaccharide from P. multiflorum is a linear α-1,4-glucan with potent immunomodulatory activity, which would be potentially developed as an effective drug.
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To recognize the potential medicinal value of the Dendrobium sonia, polysaccharide (DSP) was extracted, purified, and investigated for its immunomodulatory activity. In vitro, DSP was shown to enhance the viability (MTT assay) and phagocytosis of macrophages. In cyclophosphamide-induced immunosuppressed mice, DSP increased serum levels of TNF-α, IL-6 and IFN-γ (enzyme-linked immunosorbent assay, ELISA), and ameliorated the imbalance of the community of gut microbiota as detected by 16S ribosomal RNA gene sequencing. These results suggest that DSP might be beneficial for patients under immunosuppressed conditions.
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Oviductus ranae (OR) is a traditional Chinese medicine, which was first recorded in the Compendium of Materia Medica in the Ming Dynasty. OR contains high amounts of proteins and elicits therapeutic effects on neurasthenia, insomnia, and respiratory symptoms, which are related to oxidative stress and immunodeficiency. This study aimed to obtain the potential of OR for the development of functional food possessing antioxidant and immune-enhancement functions in the same dose. In antioxidant evaluation, OR can significantly decrease malondialdehyde and protein carbonyls (P < 0.05, P < 0.01) and significantly increase total superoxide dismutase and glutathione in a dose-dependent manner (P< 0.05, P < 0.01) against ethanol-induced oxidative stress in mice at 0.1, 1.0, and 3.0 g/kg BW. In immunomodulatory evaluation, OR could significantly enhance the phagocytosis of liver macrophages (P < 0.05, P < 0.01), delayed-type hypersensitivity response (P < 0.05, P < 0.01), hemolytic activity (P < 0.05), antibody-producing cells (P < 0.05), and natural killer cell activity (P < 0.05) in the same dose range described in antioxidant evaluation compared with those in the normal control. OR slightly influenced lymphocyte proliferation, peritoneal macrophage phagocytosis, and immune organ indices in mice. Thus, 3.0 g/kg BW OR showed potential for the development of functional food with antioxidant and immune-enhancement activities
Subject(s)
Animals , Male , Mice , Medicine, Chinese Traditional/instrumentation , Antioxidants/analysis , Functional Food/analysis , ImmunomodulationABSTRACT
Objective To investigate the physicochemical properties and immunomodulatory activities of crude polysaccharides and their fractions from Sophorae Flavescentis Radix. Methods The crude polysaccharide (SFP-100) was obtained successively by boiling Sophorae Flavescentis Radix in water, ethanol precipitating, dialyzing and freeze drying. SFP-100 was separated withDEAE-cellulose column to obtain three fractions, and these fractions were fruther separated with Sephadex G-100 column to obtain their sub-fractions. The sugar content was determined by phenol-sulfuric acid method, the molecular distribution was determined with gel filtration chromatography, and the monosaccharide composition was analyzed with capillary electrophoresis after PMP derivatization. The immunobiological activities were estimated by measuring the proliferation of mouse spleen lymphocytes as well as the IFN-secretion in mouse splenocytes and the TNF-α secretion in mouse peritoneal macrophages. Results The yield of SFP-100 from Sophorae Flavescentis Radix was 4.83％ and sugar content was 71.62％. SFP-100 was separated into three fractions SFP-100-A, SFP-100-B and SFP-100-C, whose yields were 3.5％, 25.6％ and 16.7％, and the sugar content was 85.99％, 72.09％ and 24.30％, respectively.The monosaccharide composition and their molar ratio for SFP-100-A, SFP-100-B and SFP-100-C were Ara∶Glc∶Gal=7.16∶91.02∶1.82, Xyl∶Ara∶Glc∶Rha∶Gal∶GalA=0.05∶1.00∶0.85∶0.04∶0.35∶0.43, and Xyl∶Ara∶Glc∶Rha∶Gal∶GlcA∶GalA=0.20∶1.00∶0.33∶0.36∶0.45∶0.55∶14.37, respectively. SFP-100-B was further separated into two sub-fractions SFP-100-B-a and SFP-100-B-b with Sephadex G-100, and the other two sub-fractions SFP-100-C-a and SFP-100-C-b were also obtained with the same Sephadex G-100 from SFP-100-C. SFP-100-B-a showed a main wide peak, which had the relative molecular weight 1.02×105 and monosaccharide composition Ara∶Glc∶Rha∶Gal = 1.00∶0.06∶0.02∶0.29. Meanwhile, the relative molecular weight and monosaccharide composition were 5.43×104 and Xyl∶Glc =1.00∶3.81 for the main peak of SFP-100-C-a, and 2.75×104 and Ara∶Glc∶Rha∶Gal∶GlcA∶GalA=1.00∶2.10∶0.57∶0.74∶1.09∶33.75 for the main peak of SFP-100-C-b, respectively. The crude polysaccharides SFP-100 and its fractions, SFP-100-B and SFP-100-C, as well as the sub-fractions, SFP-100-B-a, SFP-100-B-b and SFP-100-C-a, increased the proliferation of spleen cells, all in a dose-dependent manner. Further, SFP-100 could improve the secretion of IFN-γ and TNF-α in spleen cells, while SFP-100-B, SFP-100-C, SFP-100-B-a and SFP-100-B-b could stimulate the secretion of IFN-γ. Conclusion The crude polysaccharides of Sophorae Flavescentis Radix and their fractions showed a good immunomodulatory activity, which may be related to the clinical use of Sophorae Flavescentis Radix for the anti-HBV and anti-inflammatory therapy.
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Objective To extract, separate, and purify polysaccharide from Ganoderma lucidum with alkali from the residue after water extraction, characterize the basic physicochemical properties and structural features in detail, and study the immunomodulatory activity in vitro. Methods The polysaccharide LZJ-015 was isolated and purified from the dry fruiting bodies of G. lucidum by alkaline extraction and ethanol precipitation following Q-Sepharose Fast Flow ion exchange chromatography column. Monosaccharide composition and molecular weight of LZJ-0.15 was analyzed by high performance liquid chromatography (HPLC) with PMP precolumn derivatization and high performance gel permeation chromatography-multiple angle laser (HPGPC-MALLS), respectively. The detailed structure of polysaccharide LZJ-0.15 was characterized by 1H-NMR, 13C-NMR, 1H-1H COSY, and 1H-13C HSQC spectrum. The immunomodulatory activity of G. lucidum polysaccharide was test by RAW264.7 cells phagocytose neutral red experiment. Results The molecular weight, molecular radius, and Mw/Mn of LZJ-0.15 were determined to be 24 700, 46.6 nm, and 1.019, respectively. The monosaccharide composition was confirmed to be mainly composed of glucose (92.3%). LZJ-0.15 was →3) Glc (β1→and→6) Glc (β1→linked glucan indicated by NMR spectrum. Moreover, G. lucidum polysaccharide exhibited good immunomodulatory activity in our study. Conclusion G. lucidum polysaccharide LZJ-0.15 from alkaline extraction showed better immune activity than the polysaccharide extracted from water, which would be potentially developed as an effective immunomodulatory agent.
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We aimed at analyzing the structure of extracellular polysaccharide A from Grifola frondosa (EXGFP-A) and testing its immunomodulatory activity. Structural analysis shows that EXGFP-A was a contained α-D-glucoside bond and pyranose ring. GC analysis reveals that EXGFP-A was mainly composed of rhamnose, arabinose, xylose, mannose, glucose, galactose, by the molar ratio of 0.28:0.31:0.30:0.06:7.98:0.61. The results of MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay indicates when EXGFP-A was at a concentration of 80 μg/mL and treatment time of 48 h, RAW264.7 cells proliferation index reached a maximum of 137.5%. Meanwhile, the AO staining showed that EXGFP-A activated RAW264.7 cells and improved the level of intracellular nucleic acid metabolism. In addition, in a certain range of concentration, EXGFP-A was able to increase the release of NO in RAW264.7 cells, and upregulate the mRNA expression of immunological factor TNF-α, IL-1β, IL-6, IL-12, IFN-γ and iNOS of RAW264.7 cells. Our results confirm that EXGFP-A had immunomodulatory activity. Our findings provided scientific basis for the structural analysis and application of Grifola frondosa polysaccharide.
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Animals , Mice , Cytokines , Metabolism , Grifola , Chemistry , Nitric Oxide Synthase Type II , Metabolism , Polysaccharides , Allergy and ImmunologyABSTRACT
In the present work, Guduchi Ghana (concentrated form of aqueous extract of Guduchi) was prepared according to the method described in classical text – Sidhdha Yoga Samgraha and the other sample of aqueous extract was purchased from the market for the assessment of the immunomodulatory activity. It was done by haemagglutination antibody titre method for humoral immunity and footpad swelling method for cell mediated immunity on wistar albino rats. Results of present studies suggest that Guduchi Ghana prepared by classically was found to possess significant immunostimulatory action on immune system but market sample of it exhibited significant immunosuppression effect in dose dependent manner when compare with control group at a dose of 50 mg/kg orally.
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Tinospora cordifolia is used in Ayurveda as "Rasayanas" to improve the immune system and the body resistance against infections. Polysaccharides are the main constituents which are considered to be responsible for immune enhancement. In this study, immunomodulatory activity of three polysaccharide enriched fractions was evaluated using the polymorphonuclear leukocyte function test. Sugar composition was determined by GC-MS analysis of the derivatised fractions. The active polysaccharide fractions mainly constitute glucose, fructose and arabinose as monomer units.
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Withania somnifera (L) Dunal is a well known Indian medicinal plant widely used in the treatment of many clinical conditions in India. It is an important drug commonly known as Asgand which has been used either single or in combination with other drugs in Unani as well as Ayurvedic system of medicine for centuries. It has been described by Dioscorides (78 AD) in his book “Kitab-ul-Hashaish”. Asgand consists of the roots of Withania somnifera which has various therapeutic actions such as anti-inflammatory (Muhallil-e-Warm), sedative (Musakkin), alterative (Muaddil) and aphrodisiac (Muqawwi-e-Bah). Keeping in view the medicinal properties of Withania somnifera Dunal (Asgand), an attempt has been made in this review paper to explore various dimensions of the drug including phytochemical and pharmacological studies carried out on this drug.
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The immunomodulatory activity of Shirishavaleha prepared from two different parts of Shirisha (Albizia lebbeck Benth), i.e., Twak (Bark) and Sara (Heartwood) as main ingredients was evaluated for humoral antibody formation and cell-mediated immunity in established experimental models. The study used Wistar rats of either sex weighing 200 ± 40 g, while the test drug was administered orally at a dose of 1.8 g/kg. Hemagglutination titer and body weight were recorded to assess effects on humoral immunity; immunological paw edema was assessed for cell-mediated immunity. Shirishavaleha prepared from heartwood shows significant enhancement in antibody formation, attenuation of body weight changes, and suppression of immunological paw edema, while Shirishavaleha prepared from bark shows weak immunomodulatory activity. The study therefore concludes that Shirishavaleha prepared from heartwood has significant immunomodulatory activity.
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Tridax procumbens L., Asteraceae, has been extensively used in Ayurvedic system of medicine for various ailments. Previous studies on the extracts of T. procumbens revealed remarkable immunomodulatory activity of TPEIF (T. procumbens ethanol insoluble fraction) extract. The dried methanol extract of T. procumbens was dissolved in distilled water, and then fractioned by re-extracting with chloroform, ethyl acetate and n-butanol subsequently. Immunomodulatory activities of these fractions were determined in vivo. The amounts of total phenolic compounds were also determined. Ethyl acetate and n-butanol fractions showed the significant immunomodulary activity. However, the ethyl acetate fraction exhibited the highest total phenolic content. Therefore, ethyl acetate fraction was subjected to further separation by chromatographic methods. Two phytochemicals SA-3 and SA-4 were obtained by repeated purification in sufficient amount to screen them for the immunomodulatory activity by the in vivo models i.e. neutrophil adhesion and delayed type hypersensitivity. In addition, the n-butanol fraction was subjected to silica gel column chromatography (CC); SA-6 was isolated from it. Mice were treated with two doses of SA-3, SA-4 and SA-6 (2 and 4 mg/kg) for fifteen days. Immune responses to T-dependent antigen SRBCs were observed using parameters like DTH and Neutrophil adhesion. Overall, SA-4 and SA-6 showed dose relative immunostimulatory effect on in vivo immune functions in mice. From these results, it can be suggested that these compounds may be used as potential immunostimulators. The structures of isolated phytochemicals were determined by UV, IR, NMR, and MS spectroscopic methods.
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To study the immunomodulatory activity of saline extracts of leaves of Aloe vera Linn. (Family: Liliaceae) on the albino mice. The saline extract of leaves of Aloe vera was administered orally according to their body weight in mice. The assessment of immunomodulatory activity on specific and nonspecific immunity was studied by administration of test extract. The method of pyrogallol induced immunosupression was employed with slight modification to study the immunomodulatory potential of the extract. Humoral antibody response to SRBC measurement of antibody titer by haemagglutination reaction was done and cellular immune response (Foot pad reaction test) the edema was induced in the right paw of mice by injecting SRBC (0.025x109 cells) in the sub planar region. Pyrogallol-induced suppression of humoral as well as cell mediated immune response was significantly attenuated by daily oral treatment with saline extract of Aloe vera. Vitamin E treated group exhibited similar attenuation of the suppression in immune responses. Aloe vera extract at the dose of 100 mg/kg was found to suppress delayed type hypersensitivity reaction induced by SRBCs in mice. As evidenced by marked increase in haemagglutination titers in mice was also observed. The study demonstrates that A. vera triggers both specific and non-specific responses to a greater extent. The study comprised the acute toxicity and preliminary phytochemical screening of A. vera. From the results obtained and phytochemical studies the immunostimulant effect of Aloe vera could be attributed to the alkaloids content.
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Rutaceae is a taxon with species very well distributed in Brazilian semi-arid area, commonly used in folk medicine. Species from this genus have diverse biological activity described in literature. In this work, immunomodulatory and bactericidal activity are described for chloroform and ethyl acetate extracts of three of them (Esenbeckia grandiflora Mart., Pilocarpus spicatus A.St.-Hil. and Galipea simplicifolia Schult.). Initially all the samples had their cytotoxicity evaluated, aiming to determine the LC50. The immunomodulatory potential was evaluated in cultures of murine splenocytes stimulated or not with concanavalin A and in a mixed lymphocyte reaction (MLR) using splenocytes from BALB/c (H-2d) mice immunized with splenocytes from C57Bl/6 (H-2b) mice. Four samples had higher values of lymphoproliferation inhibition in concanavalin A-stimulated cultures and were evaluated in MLR. The antibacterial activity of extracts was also evaluated and the minimal inhibitory concentrations (MIC) for two active samples were 1.0 and 5.0 mg/ml for extracts from Esenbeckia grandiflora Mart. and Galipea simplicifolia Schult., respectively. Thus, our results reinforce data of literature relating biological activity for many species of the Rutaceae family and encourage studies with these species aiming to discover active compounds, candidates to new medicines.
A família Rutaceae apresenta espécies vegetais muito bem distribuídas no Semi-Árido Brasileiro e comumente usadas em medicina popular. Espécies dessa família tem diversas atividades biológicas descritas na literatura. Neste trabalho, atividades imunomoduladora e bactericida são descritas para o extrato acetato de etila e clorofórmico de três espécies da família (Esenbeckia grandiflora Mart., Pilocarpus spicatus A.St.-Hil. e Galipea simplicifolia Schult.). Todas as amostras foram inicialmente avaliadas quanto à sua citotoxicidade, com objetivo de determinar a LC50. O potencial imunomodulador foi avaliado em culturas de esplenócitos murinos estimulados ou não com concanavalina A e também em reação mista linfocitária (RML), usando também esplenócitos de camundongos BALB/c (H-2d) imunizados com esplenócitos de camundongos C57Bl/6 (H-2b). Quatro amostras tiveram os mais elevados valores percentuais de inibição da proliferação de linfócitos ativados pela concanavalina A e foram avaliados em RML. A atividade antibacteriana dos extratos foi também avaliada e a concentração mínima inibitória (CMI) para duas amostras ativas foi de 1.0 e 5.0 mg/mL, respectivamente para as espécies Esenbeckia grandiflora Mart. and Galipea simplicifolia Schult. Assim, os dados aqui apresentados reforçam informações da literatura científica relacionados à atividade biológica para muitas espécies da família Rutaceae e incentivam outros estudos com estas visando descobrir substâncias ativas, potenciais candidatas a novos fármacos.
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The family Boraginaceae is widely distributed in Brazil and in the Northeastern region some species are popularly used to treat symptoms of rheumatism, painful menstruation and dyspepsia. In this work we studied Cordia superba Cham. and C. rufescens A. DC., native from Brazilian Semi-arid region, in order to investigate their immunomodulatory activity. Six extracts were prepared from aerial parts of C. superba and C. rufescens. The cytotoxicity was evaluated using splenocytes from BALB/c mice. The immunomodulatory activity was determined by in vitro assays using activated mouse macrophages and lymphocytes. Peritoneal macrophages obtained from BALB/c mice were stimulated with IFN-gamma and LPS in the presence/absence of the samples. The NO production was measured indirectly through Griess method. Three samples inhibited the production of nitric oxide in values near 50 percent at a concentration of 100 µg/mL. To evaluate the effects of the extracts on lymphocytes, splenocytes from BALB/c mice were incubated with the samples and concanavalin A. Proliferation inhibition was determined by analysis of ³H-thymidine uptake. Samples from the two species had a strong inhibitory activity on lymphocyte proliferation and IL-2 production. Two chloroform extracts prepared from aerial parts of C. rufescens had the lowest IC50 values (7.6 and 11.0 µg/mL).
A família Boraginaceae é amplamente distribuída no Brasil e na região nordeste algumas espécies são usadas popularmente no tratamento de reumatismo, dores menstruais e dispepsias. Neste trabalho foram estudadas as espécies Cordia superba Cham. and C. rufescens A. DC., nativas da região semi-árida brasileira, objetivando investigar a atividade imunomoduladora. Seis extratos foram preparados a partir de partes aéreas das espécies. A citotoxicidade foi avaliada usando culturas de esplenócitos de camundongos BALB/c. A atividade imunomoduladora foi determinada por ensaios in vitro usando macrófagos e linfócitos murinos ativados. Macrófagos peritoneais obtidos de camundongos BALB/c foram estimulados com IFN-gama and LPS na presença/ausência das amostras. A produção de NO foi medida indiretamente através do método de Griess. Três amostras inibiram a produção de NO em valores próximos a 50 por cento (100 µg/mL). Os efeitos das amostras sobre os linfócitos foram avaliados cultivando esplenócitos de camundongos BALB/c em presença destas amostras e de concanavalina A. A proliferação foi determinada pela análise da incorporação de ³H-tritiada. Amostras de duas espécies apresentaram uma forte atividade inibidora sobre a proliferação de linfócitos e sobre a produção de IL-2. Dois extratos clorofórmicos (partes aéreas de C. rufescens) tiveram os menores valores de IC50 (7,6 and 11,0 µg/mL).