Limonium duriusculum (de Girard) Kuntze Exhibits Anti-inflammatory Effect Via NF-κ B Pathway Modulation

HIGHLIGHTS L. duriusculum n-BuOH extract reduces inflammatory responses both in vitro and in vivo. L. duriusculum n-BuOH extract inhibits NF-κB-dependent transcriptional responses. L. duriusculum n-BuOH extract decreases the expression of TNF-α and IL-6 genes.

Abstract Limonium duriusculum is used in folk medicine to treat inflammatory disorders and has gained attention due to its richness in apigenin. The present investigation was performed to evaluate and confirm its anti-inflammatory properties, in cell lines and animal models. The potential anti-inflammatory properties of n-butanol (n-BuOH) extract of L. duriusculum (BEL) and isolated apigenins were examined on NF-κB transcriptional activity in TNFα- or LPS-stimulated cells, and on in vivo acute inflammatory models (carrageenan induced paw edema and peritonitis). BEL treatment was able to inhibit the activity of an NF-κB reporter gene in HCT116 cells both in the absence and in the presence of exogenous TNFα, used as NF-κB pathway inducer. This anti-inflammatory effect was even more potent compared to Apigenin (APG1) and was confirmed using monocyte-derived THP-1 cells treated with LPS to stimulate NF-κB-dependent transcription of IL-6 and TNFα mRNAs. Apigenin7-O-β-(6''-methylglucuronide) (APG2) was instead inactive both in HCT116 and THP-1 cells. BEL (oral, 200 mg/kg) led to paw swelling inhibition, vascular permeability and peritoneal leukocyte and PN migration diminution. Apigenins (intraperitoneal, APG1, APG2: 20 mg/kg) also evoked a significant anti-edema effect, early vascular permeability and leukocyte influx reduction. Collectively, this study demonstrates for the first time the effectiveness of L. duriusculum to inhibit NF-κB-dependent transcriptional responses in HCT116 and THP-1 cells. In vivo studies also established that L. duriusculum possesses a potential anti-inflammatory effect, confirm its traditional, empirical use, that could be attributed to its richness in apigenin.

Avaliação da atividade imunomoduladora das pectinas in natura e modificadas obtidas de goiabas / Evaluation of immunomodulatory activity of non-modified and modified pectins from guava fruit

As pectinas presentes nas frutas, assim como sua versão modificada, estão entre as biomoléculas mais promissoras no campo da imunomodulação. Este estudo teve como objetivo avaliar o potencial imunomodulador de pectinas obtidas de goiabas verdes e maduras, bem como suas versões modificadas. As goiabas cv. Tailandesas foram avaliadas ao longo do amadurecimento, sendo acompanhadas as alterações da cor da polpa e casca, firmeza, produção de etileno e respiração, perda de massa e o teor de açúcares solúveis. Após a caracterização dos frutos, foram extraídas as pectinas e estas foram caracterizadas quanto ao conteúdo monossacarídico, peso molecular, presença de oligossacarídeos e grau de esterificação. As pectinas in natura de goiaba madura e verde, bem como a versão modificada desta última apresentaram frações de alto e baixo peso molecular, alta proporção de ácido galacturônico e alto teor de esterificação. Por outro lado, as pectinas modificadas derivadas de goiaba madura apresentaram maior desesterificação, com perda de frações de menor peso molecular, menor proporção de ácido galacturônico e baixo teor de esterificação. As pectinas foram incubadas com células THP-1 e RAW 264.7, e apesar da alta viabilidade celular e ausência de efeito citotóxico, resultou em expressiva produção de espécies reativas de oxigênio. De modo geral, as pectinas in natura de goiaba verde e madura, e pectina modificada de goiaba verde promoveram estímulo da produção de citocinas diversas, em especial inflamatórias, como IL-1ß, IL-12, CCL5, CXCL10 e CXCL9, para células THP-1 e IL-10 (antiinflamatória), TNF-α e MCP-1, demonstrando seu potencial imunomodulador, já para células RAW 264.7 as pectinas estimularam a produção de IL-10, TNF-α e MCP-1, demonstrando seu potencial imunomodulador. A pectina modificada derivada da goiaba madura não promoveu a indução significativa de nenhuma citocina. Estes resultados sugerem que as pectinas obtidas a partir de goiabas têm potencial imunomodulador e devem ser estudadas em outros modelos celulares e / ou em concentrações mais altas e modelos in vivo, para que esses benefícios possam realmente ser comprovados

The pectin present in fruits and their modified version are the most promising biomolecules in the immunomodulatory field. This study aimed to evaluate the immunomodulatory potential of pectins obtained from unripe and ripe guavas (Psidium guajava L.) cv. Thailandesa as well as their modified versions. The guavas were characterized during ripening regarding cell wall solubilization, sugar content, firmness, mass loss and ethylene production rate and respiration during 10th following harvest. After fruit characterization, pectins were extracted and characterized for monosaccharide content, molecular weight, presence of oligosaccharides and degree of esterification. Pectins from unripe and ripe guava as well as the modified versions of pectins had high and low molecular weight fractions, a high proportion of galacturonic acid and high esterification content. On the other hand, modified pectins derived from ripe guava showed higher de-esterification, with loss of lower molecular weight fractions, a lower proportion of galacturonic acid and low esterification content. The pectins were incubated with THP-1 and RAW 264.7 cells, and despite the high cell viability and absence of cytotoxic effect, the treatment resulted in the expressive production of reactive oxygen species. In general, pectins from ripe and unripe guava and modified pectin from unripe guava stimulated the production of diverse cytokines, especially inflammatory cytokines, such as IL-1ß, IL 12, CCL5, CXCL10, and CXCL9, for THP-1, while IL-10 (anti-inflammatory), TNF-α and MCP-1 were stimulated in RAW 264.7 cells, demonstrating their immunomodulatory potential. Modified pectin derived from ripe guava did not promote any significant induction of the cytokines investigated. These results suggest that pectins obtained from guavas have immunomodulatory potential and deserve a more in-depth investigation using other cellular models and/or use of higher concentrations and in vivo models tests so their immunomodulatory benefits can be prove

Effectual components of Polygonum ciliinerve protects against Staphylococcus aureus infection with immunomodulatory functions in C57BL/6 mice

Abstract Purpose: To investigate the efficacy and mechanisms of root tuber of Polygonum ciliinerve (Nakai) ohwi (rPC) which has been used to treat bacterial infection in traditional Chinese medicine. Methods: With the mouse model of Staphylococcus aureus (S. aureus) pneumonia, the phenotype of rPC treated mice, including body weight, mortality, lung slices and bacterial burden were evaluated. Furthermore, inflammatory factors in bronchoalveolar lavage (BAL) were determined by ELISA and the distribution of T cells in lung was assessed by immunofluorescence assay. Results: rPC treatment could dose-dependently reduce weight loss and mortality in S. aureus-infected mice. Upon 10 mg/ml rPC treatment, S. aureus-infected mice showed about 8 grams increase in body weight (P<0.001) and 50% enhancement in mortality. The integrity of lung tissue and bacterial burden were also improved by rPC treatment. Moreover, rPC was found to modulate the immune response in infection. Conclusion: rPC has therapeutic potential for S. aureus infections and pneumonia with immunomodulatory functions.

Extractos vegetales de tres especies del género Baccharis inducen la proliferación de células mononucleares humanas / Plant extracts of three species of the genus Baccharis induce human mononuclear cells proliferation

Las plantas de uso en medicina tradicional constituyen una fuente importante de compuestos con actividad inmunomoduladora; entre ellas las especies del género Baccharis, conocidas popularmente como "Jaguareteka´a" en nuestro país, son ampliamente empleadas. En este estudio se evaluó la actividad inmunomoduladora de extractos metanólicos de tres especies del género Baccharis (B. trimera, B. notosergilay B. punctulata) sobre la proliferación de células mononucleares humanas de sangre periférica. Los extractos de las tres especies estudiadas estimularon la proliferación de las células mononucleares. Específicamente, el extracto de B. notosergila estimuló la proliferación celular a todas las concentraciones probadas (5, 10, 25 y 50 µg/mL), mientras que los extractos de B. trimera y B. punctulata mostraron este efecto a 5 y 10 µg/mL. Además, por presentar mayor inducción de la proliferación, se realizó un fraccionamiento con diferentes solventes del extracto metanólico de B. notosergila y B. punctulata. La fracción de acetato de etilo de ambos extractos vegetales aumentó la proliferación celular, sugiriendo que compuestos de polaridad media son los responsables de esta actividad. Estos resultados demuestran que los extractos de B. trimera, B. notosergila y B. punctulata poseen actividad inmunomoduladora sobre células mononucleares humanas y servirán de base a otros estudios para determinar el o los componentes activos de los extractos sobre el sistema inmune(AU)

Plants used in traditional medicine are an important source of compounds with immunomodulatory activity. Species of the genus Baccharis, popularly known as "Jaguareteka'a" in our country, are used in folk medicine for the treatment of liver, gastrointestinal, inflammatory and infectious diseases. In this study, we evaluated the immunomodulatory activity of methanolic extracts of three species of the genus Baccharis (B. trimera, B. notosergila and B. punctulata) on the proliferation of human peripheral blood mononuclear cells. Extracts of the three species studied stimulated the proliferation of mononuclear cells. The extract of B. notosergila stimulated cell proliferation at all concentrations tested, while extracts of B. trimera and B. punctulata stimulated at 5 and 10 µg/mL. In addition, we carried out a separation with different solvents of the methanolic extract of B. notosergila and B. punctulata. The ethyl acetate fraction of both plant extracts induced the proliferation of immune cells. These results show that the extracts of B. trimera, B. notosergila and B. punctulata had immunomodulatory activity on human mononuclear cells. Future work will be required to identify the components responsible for the activity on the immune system(AU)

Cordyceps sinensis promotes immune regulation and enhances bacteriostatic activity of PA-824 via IL-10 in Mycobacterium tuberculosis disease

PA-824 is a novel bicyclic nitroimidazole anti-tuberculosis (TB) drug. Cordyceps sinensis (Berk.) Sacc. (CS) was proven to be a good immunomodulatory compound. This research aimed to investigate the effect of CS on PA-824 in Mycobacterium tuberculosis (M.tb) infected mice (female CBA/J mice, 6 to 8 weeks of age and 20±2 g of weight). Mice were randomly assigned to 4 groups: PA-824, CS, PA-824+CS, and control. To verify the effect of PA-824 and CS on M.tb, after drug administration, mice lungs were harvested and bacterial colony formations were measured. Cells were isolated from infected lungs and spleens to analyze the percentage of CD4+ T cells (CD11a positive). Lung cells were cultured to detect the secretion of interferon-γ (IFN-γ) and interleukin-10 (IL-10) by ELISA. IFN-γ and IL-10 double-positive CD4+ cells in peripheral blood were measured by flow cytometry. The expression levels of IL-2 and IL-10 in mice lungs were analyzed by real-time PCR and western blot. Results showed that PA-824 combined with CS led to the lowest lung colony-forming units (CFU) counts among treated groups. Furthermore, this beneficial outcome might be associated with the decreased CD11a on CD4+ cells in mice lungs and spleens. Moreover, the suppressed secretion of IFN-γ and IL-10, and IL-10 expressions, as well as the decreased IFN-γ and IL-10 double-positive CD4+ cells in blood, could also be associated with the positive effect. However, no significant effect on IL-2 production was found. The combination of PA-824 and CS had more effective bacteriostatic and immunomodulatory effects on M.tb infected mice than PA-824 alone. In conclusion, CS has the potential to be an effective adjuvant in TB treatment.

Immunomodulatory activity of Tityus serrulatus scorpion venom on human T lymphocytes

Background Tityus serrulatus scorpion venom (TsV) contains toxins that act on K + and Na + channels and account for the venoms toxic effects. TsV can activate murine peritoneal macrophages, but its effects on human lymphocytes have been poorly investigated. Considering that lymphocytes may play an important role in envenomation, we assessed whether TsV affects the expression of phenotypic (CD3, CD4, and CD8) and activation (CD69, CD25, and HLA-DR) markers, cell proliferation, and cytokine production in peripheral blood mononuclear cells. Methods Cytotoxicity of TsV was evaluated via the MTT assay. Cell proliferation, expression of phenotypic and activation markers, and release of cytokines were assessed using flow cytometry, after treatment with non-cytotoxic concentrations of TsV. The combined use of carboxyfluorescein diacetate succinimidyl ester and monoclonal antibodies against phenotypic and activation markers enabled us to simultaneously assess cell proliferation extent and cell activation status, and to discriminate among cell subpopulations. Results TsV at concentrations of 25 to 100 g/mL were not cytotoxic towards peripheral blood mononuclear cells. TsV did not induce significant changes in lymphocyte subpopulations or in the expression of activation markers on CD4 + and CD8 + T cells. TsV inhibited the phytohemagglutinin-stimulated lymphocyte proliferation, particularly in the CD8 + CD25 + T lymphocyte subset. TsV alone, at 50 and 100 g/mL, did not induce peripheral blood mononuclear cell proliferation, but elicited the production and release of IL-6, a proinflammatory cytokine that plays an important role in innate and adaptive immune responses. Conclusions TsV is a potential source of molecules with immunomodulatory action on human T lymphocytes.

Efeito anti-inflamatório dos macrolídeos em doenças pulmonares da infância / Anti-inflammatory effects of macrolides in childhood lung diseases

Os macrolídeos são fármacos com efeitos antimicrobianos especialmente contra patógenos intracelulares. Vários estudos têm demonstrado possíveis efeitos anti-inflamatórios dos macrolídeos. Esses medicamentos inibem a produção de algumas interleucinas e podem reduzir a inflamação neutrofílica pulmonar. Ensaios clínicos têm demonstrado efeitos benéficos dos macrolídeos em diversas doenças pulmonares crônicas. O objetivo deste estudo foi revisar os dados recentes da literatura médica sobre os efeitos anti-inflamatórios dos macrolídeos nas doenças respiratórias da infância, através da pesquisa da base de dados Medline (PubMed) dos seguintes termos em inglês: "macrolide and cystic fibrosis"; "macrolide and asthma"; "macrolide and bronchiolitis obliterans"; e "macrolide and acute bronchiolitis" Foram selecionados artigos publicados em revistas científicas internacionais entre 2001 e 2012. Estudos clínicos e evidências in vitro comprovam o efeito anti-inflamatório dos macrolídeos em doenças respiratórias. Alguns ensaios clínicos demonstram benefícios na administração de macrolídeos em pacientes com fibrose cística; porém, o risco de resistência bacteriana deve ser considerado na análise desses benefícios. Tais benefícios são controversos em outras doenças respiratórias, e seu uso rotineiro não está indicado. Mais estudos clínicos controlados são necessários para avaliar a eficácia desses medicamentos como anti-inflamatórios. Dessa forma, poderemos definir melhor os benefícios dos macrolídeos no tratamento de cada uma das situações clínicas especificadas.

Macrolides are drugs that have antimicrobial effects, especially against intracellular pathogens. Various studies have shown that macrolides might also have anti-inflammatory effects. Macrolides inhibit the production of interleukins and can reduce pulmonary neutrophilic inflammation. Clinical trials have demonstrated beneficial effects of macrolides in various chronic lung diseases. The objective of this study was to review recent data in the medical literature on the anti-inflammatory effects of macrolides in childhood lung diseases by searching the Medline (PubMed) database. We used the following search terms: "macrolide and cystic fibrosis"; "macrolide and asthma"; "macrolide and bronchiolitis obliterans"; and "macrolide and acute bronchiolitis". We selected articles published in international scientific journals between 2001 and 2012. Clinical studies and in vitro evidence have confirmed the anti-inflammatory effect of macrolides in respiratory diseases. Some clinical trials have shown the benefits of the administration of macrolides in patients with cystic fibrosis, although the risk of bacterial resistance should be considered in the analysis of those benefits. Such benefits are controversial in other respiratory diseases, and the routine use of macrolides is not recommended. Further controlled clinical trials are required in order to assess the efficacy of macrolides as anti-inflammatory drugs, so that the benefits in the treatment of each specific clinical condition can be better established.

Immunomodulating effect of ampicillin and cephalosporin in sheep vaccinated with sheep poxvirus vaccine

In this study we use 20 sheep divided into four groups [one group was injected with the vaccine, the second was taken the vaccine and ampicillin, the third group was taken the vaccine and cephalosporin and the fourth act as a control]. The immunity was measured humoral immunity to sheep pox virus vaccine by neutralization test and cell mediated immunity by lymphocyte transformation assay as well as phagocytosis. The results revealed that an increase of the immune status in group injected with antibiotics with vaccine than that received the vaccine only and than the control group. There were an increase of lymphocyte transformation test, neutralization test and phagocytosis. It could be concluded that ampicillin and cephalosporin directly increase the immune status in sheep vaccinated with sheep poxvirus vaccine

Prophylactic effect of Bovine lactoferrin against acute toxoplasmosis in immunocompetent and immunosuppressed mice

This study evaluated the immune-potentiating effect of administrating bovine Lactoferrin [LF] to immunocompetent [IC] and immunosuppressed [IS] mice prior to infection with tachyzoites of T. gondii RH strain. Mice were IS with cyclophosphamide. LF was given in seven of them as oral doses on alternate days. Immunological and parasitological assessments showed that LF induced statistical significance comparable resistance against acute toxoplasmosis in IC and IS mice. This was verified by elevated splenic CD4[+] T lymphocytes, reduced tachyzoites' viability and infectivity, with diminished parasite burdens. So, mice mortality declined and their survival was prolonged. This indicated that LF have prophylactic efficacy against human toxoplasmosis in risky persons with alleviating immune balance

Influence of biphenyl dimethyl dicarboxylate [DDB] on social play behavior of young rats

Biphenyl Dimethyl Dicarboxylate [DDB] is a synthetic analogue of schisandrin C, a component of Fructus schisandrae. It is a widely prescribed agent for the improvement of liver function of hepatitis patients and has been recently proved to possess immunomodulatory properties. In our earlier work, DDB was shown to interfere with behavior. The present work was to assess the possible effect of DDB on the play behavior. The study was carried out on young Sprague Dawley Rats. DDB was administered orally for seven consecutive days [100mg/kg/day]. Animals were isolated for about 28hr before the day of the experiment. Each pair was placed in an aquarium glass cage where the play behavior of a pair of animals was recorded by a special Infrared Video Camera allowing super night shots, for 25 min. Three major categories were assessed, namely "play social behavior", "non-play social behavior" and "non-social behavior". DDB was found to affect the play behavior of the partner rat where the "pounce on" and "pinning" were significantly increased. Parallel changes in brain NE were found. In conclusion, DDB was shown to change the play behavior of the partner rat

Endogenous and exogenously-induced immunomodulation of tumour-host responsiveness

In spite of the availability of multiple effector mechanisms of the immune system to combat tumour growth and metastases, their impairment frequently accompanies the appearance of cancer. Factors contributing to this impairment may be related to properties of the host and/or the tumour itself and may be with respect to their origin -endogenous or exogenour. Based on the unique biological behavior of prostate cancer (PCa), and its apparent escape from immune surveillance in the presence of tumour immuno genicity, continuing investigation of endogenous and exogenous factors thought to be relevant to its pathogenesis have been made. For this purpose further studies of the suggested role of human seminal plasma (SePl) and the synthetic oestrogen, diethylstiboestrol (DES), as representative endogenous and exogenous immunomodulatory factors (IMF) of tumour-host responsiveness, together with evaluation of human prostatic tissue extracts and leuprolide (the luteinizing-hormone-releasing-hormone proposed as an alternate to DES therapy) have been made by evaluating their effect on the lytic activity of natural killer (NK) cells. SePl and prostate extracts significantly suppressed NK cell lysis. Physicochemical studies suggest SePl and prostate IMF to be associated with high and low molecular weight macromolecules; and implicate the participation of transglutaminase and prostaglandins. Comparative study of therapeutic levels of DES vs. leuprolide on NK cell lysis demonstrated significant suppression by DES vs. a negligible effect of leuprolide. Metastases are highly prevalent in PCa, and contribute significantly to its morbidity and mortality. Further knowledge of the range of effects of endogenous and exogenous IMF on effector mechanisms of tumour-host responsiveness, to include suppression of NK cells, and elucidation of their nature, may contribute toward our understanding of the unique biological behavior of tumours of the prostate, in addition to improvement in their clinical management.