ABSTRACT
Achyranthes japonica Nakai (A. japonica) is a medicinal herb found widely distributed throughout Korea. The biological activities of A. japonica are well-documented and include anti-fungal, anti-inflammatory, and immunity enhancement. The objective of the present study was to investigate the immune-related activities of A. japonica extract in dogs. The extract was acquired by ethanol extraction and purified by filtration. To examine the effect of A. japonica extract on immune cell viability, human lymphocytes, such as Jurkat T-cells and Ramos B-cells, were exposed to the extract. After treatment with the extract, the number of Ramos B-cells was increased, whereas Jurkat T-cells remained unaffected. Griess assay revealed decreased nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated mouse macrophage Raw 264.7 cells after exposure to A. japonica extract. To evaluate the in-vivo effect in dogs, feed containing A. japonica extract was provided to 8 dogs for 2 months. Blood samples were collected before, during, and after consumption of the feed. Peripheral blood mononuclear cells (PBMCs) were isolated from the blood samples and the number of T-cells and B-cells were assessed using flow cytometry with anti-dog fluorescein isothiocyanate (FITC)-conjugated CD3 and anti-dog phycoerythrin (PE)-conjugated CD21 antibodies, respectively. We observed a significant increase in the average number of B-cells in the PBMCs during ingestion of the feed containing A. japonica. In addition, enzyme-linked immunosorbent assay (ELISA) revealed a decrease in the levels of tumor necrosis factor-alpha (TNF-α), a pro-inflammatory cytokine, in 3 out of 8 dogs and increased levels of interleukin-10 (IL-10), an anti-inflammatory cytokine, in 4 out of 8 dogs. Taken together, we believe that these changes indicate that A. japonica extract is beneficial in improving the immunity of dogs by stimulating B-cells and inducing production of anti-inflammatory responses.
Achyranthes japonica Nakai (A. japonica) est une herbe médicinale retrouvée largement distribuée à travers la Corée. Les activités biologiques d'A. japonica sont bien documentées et inclus des effets antifongique, anti-inflammatoire et de stimulation de l'immunité. L'objectif de la présente étude était d'examiner les activités reliées à l'immunité d'un extrait d'A. japonica chez des chiens. L'extrait fut obtenu par extraction à l'éthanol et purification par filtration. Pour examiner l'effet de l'extrait d'A. japonica sur la viabilité de cellules immunitaires, des lymphocytes humains, tels que les cellules T Jurkat et les cellules B Ramos, furent exposés à l'extrait. Après traitement avec l'extrait, le nombre de cellules B Ramos était augmenté, alors que celui des cellules T Jurkat était inchangé. L'épreuve de Griess a révélé une diminution de production d'oxyde nitreux (NO) chez les macrophages de souris Raw 264,7 stimulés par le lipopolysaccharide (LPS) à la suite de l'exposition à l'extrait d'A. japonica. Afin d'évaluer les effets in vivo chez les chiens, de la nourriture contenant l'extrait d'A. japonica fut donnée à huit chiens pour une période de 2 mois. Des échantillons sanguins furent prélevés avant, durant et après consommation de l'aliment. Des mononucléaires du sang périphérique (PBMCs) furent isolés des échantillons sanguins et le nombre de cellules T et de cellules B fut évalué en utilisant la cytométrie de flux et des anticorps anti-CD3 de chien conjugués à l'isothiocyanate de fluorescéine (FITC) et des anticorps anti-CD21 de chien conjugués à la phycoérythrine (PE), respectivement. Nous avons observé une augmentation significative du nombre moyen de cellules B dans le PBMCs durant l'ingestion de la nourriture contenant A. japonica. De plus, une épreuve immuno-enzymatique (ELISA) a révélé une diminution des niveaux du facteur alpha nécrosant des tumeurs (TNF-α), une cytokine pro-inflammatoire, chez trois des huit chiens et des niveaux augmentés d'interleukine-10 (IL-10), une cytokine anti-inflammatoire, chez quatre des huit chiens. Pris globalement, nous croyons que ces changements indiquent qu'un extrait d'A japonica est bénéfique pour améliorer l'immunité chez les chiens en stimulant les cellules B et en induisant la production de réponses anti-inflammatoires.(Traduit par Docteur Serge Messier).
Subject(s)
Achyranthes/chemistry , Anti-Inflammatory Agents/pharmacology , Interleukin-10/blood , Lymphocytes/drug effects , Plant Extracts/pharmacology , Tumor Necrosis Factor-alpha/blood , Animals , Anti-Inflammatory Agents/chemistry , Cell Line , Cell Survival/drug effects , Dogs , Female , Gene Expression Regulation/drug effects , Humans , Interleukin-10/genetics , Interleukin-10/metabolism , Lymphocytes/physiology , Male , Mice , Nitric Oxide/metabolism , Plant Extracts/chemistry , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Pesticides are chemical substances used to kill unwanted fungi, weeds and insects. In many countries, there is currently concern regarding the adverse effects of pesticides on health. It has been reported that pesticides may cause cancer, respiratory diseases, organ diseases, system failures, nervous system disorders and asthma, which are closely connected with immune disorders. Therefore, this study reviewed the immunotoxicity of pesticides that are currently used or prohibited from being used, especially their effects on leukocytes such as T cells, B cells, NK cells and macrophages. These immune cells play crucial roles in innate and adaptive immune systems to protect hosts. Pesticides are known to have possible toxicological modes of action to induce oxidative stress, mitochondrial dysfunction, and endoplasmic reticulum (ER) stress in living organisms. According to previous studies, pesticides such as atrazine (ATR), organophophorus (OP) compounds, carbamates, and pyrethroids were shown to inhibit the survival and growth of leukocytes by inducing apoptosis or cell cycle arrest and interfering with the specific immunological functions of each type of immune cells. These results suggest the immunotoxicity of pesticides toward specific immune cells. To substantiate the overall immunocompromised effects of pesticides, there is a need to collect and thoroughly analyze additional information regarding other immunological toxicities.
Subject(s)
Immune System/physiopathology , Pesticides/adverse effects , Animals , Humans , Immune System/drug effectsABSTRACT
Fludioxonil is fungicide used in agriculture, which is present in fruits and vegetables. In this study, the effects of fludioxonil on human immune cell viability, apoptosis, cell cycle arrest, and mitochondrial membrane potential were examined in human immune cells, such as Jurkat T cells and Ramos B cells. To examine the cell viability, Jurkat T cells and Ramos B cells were treated with fludioxonil (10-9-10-5 M) for 24 h and 48 h. Water soluble tetrazolium salt assay showed that fludioxonil decreased Jurkat T cell and Ramos B cell viability. Jurkat T cell viability decreased at 24 and 48 h, but Ramos B cell viability decreased only at 48 h. JC-1 dye revealed decreased mitochondrial membrane potential in fludioxonil-treated Jurkat T cells and Ramos B cells. To evaluate apoptosis, annexin-V conjugated FITC, AF488, and propidium iodide (PI) were used and to evaluate cell cycle arrest PI was used. Apoptosis and cell cycle arrest were induced by fludioxonil (10-7-10-5 M) in the Jurkat T cells at 24 and 48 h and Ramos B cells at 48 h. Moreover, the protein levels of pro-apoptotic proteins, such as p53, BAX, and cleaved caspase 3, were increased and anti-apoptotic protein Bcl-2 was decreased by fludioxonil. Expression of the Fas receptor related to the extrinsic apoptosis pathway was increased by fludioxonil. Additionally, cyclin D1 and cyclin E1 were decreased by fludioxonil. In the present study, fludioxonil induced immunotoxicity in human T cells and B cells through apoptosis and cell cycle arrest. Therefore, the present study suggests that fludioxonil induces the cellular toxicity in immune cells.
