Resveratrol and immune system -
Irradiation protocol was approved by the Atomic Energy Commission of Syria according to the IAEA regulations and safety measures. The abdomen was opened through a midline incision; then a 25G needle was inserted in the inferior vena cava to collect blood into a citrate containing tube for the immuno-magnetic separation IMS analysis of CECs; then, the animals were sacrificed via cervical dislocation and the liver was dissected, rinsed with PBS.
Hepatocytes and Kupffer cells isolated from each liver sample were rinsed with PBS, then suspended in 1 ml of Tris-Urea-CHAPS lysing buffer 50 mM Tris.
Cl, pH 7. Supernatant was directly used to quantify the cytokines using Rat Th Cytokine panel plex kit, BioLegend ® , Cat. Briefly, 25 µl of the cleared supernatant sample was incubated with beads coated with each anti-cytokine antibodies then washed and incubated with detection antibodies before washing and analyzing.
Standard cytokine panel cocktail was utilized to build the standard curves, which were then used to calculate cytokine concentrations. BD Biosciences FCAP software was deployed to calculate cytokine concentrations.
Three replicates of each liver tissue sample were measured. Hepatocytes and Kupffer cells were isolated from Wistar rat livers according to a modified method previously described by Shulman and Nahmias Frozen liver tissue samples were brought into room temperature and rinsed with PBS, then each liver sample weighing 50 mg was minced in a sterile petri dish containing 5 ml of collagenase dissolved in Krebs Ringer Buffer 0.
Krebs Ringer Buffer recipe: Kupffer cells recovered from the band in the supernatant, and hepatocytes recovered from the pellet. Cells were then washed with PBS and used in other assays.
PCR was performed using 0. Mx3OO5P QPCR system Agilent technologies, Germany was used to quantify the expression of target genes according to the procedure described in previous works Khalil et al. Primer sequences Eurofins Genomics, Germany for the reference and target genes were:.
Total protein was extracted from isolated hepatocytes and Kupffer cells by adding 1 ml of ice-cold nondenaturing lysis buffer to 0. Cells were suspended in lysis buffer by gentle agitation for seconds with a vortex mixer set at medium speed, then the suspension was kept on ice for 15 to 30 min.
IL levels were measured using Interleukin ELISA kit Sigma Aldrich ® , St. Specimens were cut into 5 µm thick sections by cryostat SLEE MNT Cryostat Medical GmbH, Germany , then stained with hematoxylin Flukachemie Gm bHcH, Switzerland and eosine Qualikems laboratory Reagent, India according to standard histological staining procedures.
Stained liver tissues were imaged using a light microscope BX53, Olympus, Japan connected to a personal computer. Endothelial cells were isolated from the whole blood using Dynal M IgG1 immune magnetic beads Dynal AS, Oslo, Norway.
Beads were then washed three times with PBS—BSA—NaN3 to remove excess of antibodies then resuspended with buffer until use. Separation of beads rosetted with endothelial cells from the blood samples required a minimum of 2 min exposure to the magnet.
Three washes were performed to completely remove non-rosetted cells. Endothelial cells were counted using 0. The quality of images was improved by Deltapix software version 1. All experiments were repeated three times, and data were statistically analyzed using a Mann—Whitney U test and presented as the mean ± SD.
All statistical analyses were performed with GraphPad Prism 7. For endothelial cell quantitation, the results were presented as Mean cell number ± SEM. In addition, RSV administration increased IL levels significantly by approximately 2.
No alterations in IL-4, IL-5, and IL levels were observed after administrating RSV at any used dose Figures 2A, B, E. However, our results showed that IL-4, IL-5, IL-6, IL, and IL levels were significantly increased in irradiated rats compared to non-irradiated non-resveratrol treated control C group.
This increase was ~3-fold Figure 2A , 2. Figure 2 The cytokine profiles in Wistar rat liver produced by Th2 cells, which include A IL-4, B IL-5, C IL-6, D IL10, and E IL in six rat groups. Whereas IL-9, IL A and F , GM-CSF, and IL significantly increased by about 3.
ILA and GM-CSF which enhance pro-inflammatory cytokine production Brembilla et al. The same result was obtained for IL, which somehow decreased by about 0. Figure 3 The cytokine profiles in Wistar rat liver produced by Th9,Th17, and Th22 cells which include A IL-9, B ILA, C ILF, D GM-CSF and E IL in six rat groups.
IL is a very important anti-inflammatory cytokine, which maintains normal tissue homeostasis Iyer and Cheng, The results of the current study found that RSV induces the production of IL, the most significant anti-inflammatory cytokine among all measured cytokines.
In order to identify the origin of this important anti-inflammatory cytokine induced by RSV administration, we isolated the hepatocytes and Kupffer cells. RSV increased IL mRNA expression and IL levels in both hepatocytes and Kupffer cells.
This increase is further confirmed by an increase in IL10 protein levels extracted from isolated cells Figure 4B. The concentrations of IL in KCs and in HCs isolated from livers of irradiated rats pretreated with RSV increased by about 2. Figure 4 Effects of a 2 Gy single dose of whole body irradiation and RSV on IL10 relative mRNA gene expression and protein levels in Hepatocytes A , C and Kuppfer cells B, D of Wistar rats.
A histological study on liver tissues was conducted to evaluate the effects of RSV on the histology of hepatic tissue and on the number of Kupffer cells, which are the most abundant immune cells in the liver.
Figure 5 demonstrates that irradiation increased the number of Kupffer cells Figure 5B in the liver of irradiated rats compared to the non-irradiated control Figure 5A. Pretreatment with resveratrol enhanced the CEC counts in peripheral blood, which implies radioprotective effect.
Radiation exposure has been reported to induce several cytokines and growth factors in vivo and in vitro such as TNF-α, IL-1 β , IFN- γ , GM-CSF, IL-4, IL-5, IL-6, IL, IL, and IL, and TGF- β Zhang et al.
IL-2, TNF-α, and INF- γ are pro-inflammatory cytokines produced by Th1 cells and amplifying the inflammatory response Martin et al. These cytokines were upregulated by irradiation with 2 Gy dose of gamma radiation, and this result was expected as irradiation leads to upregulation of pro-inflammatory cytokines such as TNF-α, IL-1 β , INF- γ , IL-6 Shan et al.
RSV administration before irradiation resulted in a slight inhibition of TNF-α production Figure 1B. It also reduced the levels of INF- γ significantly compared to irradiated non-resveratrol treated control Figure 1C.
This is consistent with a previous study in which resveratrol could significantly reduce the levels of pro-inflammatory factors: TNF-α, IL-1 β , and IL- 6 and greatly promote the expression level of SIRT1 pathway, which is the main pathway of pro-inflammatory inhibition by RSV Schaue et al.
However, in our experiments, IL-2 generated by Th1 cells and IL-6 produced mainly by Th2 cells were the only pro-inflammatory which was upregulated by RSV in rat liver subjected to irradiation Figures 1A and 2C.
Also, it was demonstrated that mRNA and serum levels of IL-2, IL-6, and IL increased in patients post oral administration of resveratrol Multhoff and Radons, In addition, RSV was found to enhance the expression of IL-1 β and IL-6 in the peripheral blood lymphocytes Wendling et al.
These results are in accordance with our results. In fact, the enhanced production of IL-1 β and IL-6 characterizes a pro-inflammatory status contributing to T helper lymphocyte differentiation and function BaGen et al.
Thus, this increase in IL-6 and IL-2 may be involved in tissue repair and regeneration post-irradiation. Furthermore, the immune cells exposed to RSV in the vascular compartment expressing significant levels of IL-1 β or IL-6 are triggered for the adaptive immune response Malaguarnera, GM-CSF is a pro-inflammatory cytokine that acts at the interface between innate and adaptive immunity Rutz et al.
RSV also ominously reduces the production of this pro-inflammatory cytokine Figure 3D which is in agreement with the results reporting that RSV reduces the expression of GM-CSF in TNF-α activated human umbilical vein endothelial cells Wendling et al.
In fact, the Th17 cells play very important roles in autoimmune diseases, which are key initiators of pro-inflammatory responses through recruiting neutrophils and macrophages to injured tissues and via their production of IL, which in turn plays an important role in host defense against infection of extra cellular pathogens.
Moreover, Th17 cells produce IL, which, similar to IL, is beneficial to the host in many inflammatory disorders Schwager et al. However, depending on the target tissue, IL can be pathogenic due to its inherent pro-inflammatory properties, which are further enhanced when IL is released together with other pro-inflammatory cytokines, in particular IL Astry et al.
In fact, RSV, in our study prevents the link of IL and IL Thus, RSV plays a protective role against the unexpected consequences from IL and IL operating together Figures 2B, E. RSV exhibits an anti-inflammatory profile in macrophages Walle et al. Our results support this role of RSV in KCs of the liver by producing IL, which has a strong anti-inflammatory role in the immune response.
RSV enhances hepatocytes as reported in the current study to produce IL response, but as demonstrated in Figures 2D and 3 , the IL production by HCs is less important versus production of IL by KCs that might be due to the increased number of KCs by RSV effect Figure 5D exhorting KCs to produce more anti-inflammatory response mediated by IL in our study Figures 4A, B.
However, this finding indicates that RSV may promote IL mediated immune resolution as reported previously Chung, ; Bonizzi and Karin, ; Ogawa et al. Actually, this result is of a great importance and leads us to conclude that RSV effects after whole body irradiation are mainly via KCs derived-IL and not via HCs.
Moreover, IL plays a very important role in mediating host anti-inflammatory response. Therefore, identifying the cellular sources of IL in addition to the molecular mechanisms that regulate IL expression is critical to developing therapeutic strategies directed against pathology associated with impaired IL production Iyer and Cheng, In fact, these results give a great essential key role to liver KCs in immune response post whole body irradiation as a cellular and molecular target of future pharmacological molecule such as RSV.
Moreover, RSV enhances the expression of the anti-inflammatory cytokine IL-4 produced by Th2 cells Figure 2A post irradiation. In fact, this effect of RSV to enhance the anti-inflammatory response by IL-4 and IL is in accordance with a similar study in mouse bone marrow mesenchymal stem cells for therapeutic purposes reported by other groups Wang et al.
Regarding IL-5 and IL produced by Th2 cells, our data presented in Figures 2A, B, E demonstrated that these cytokines were not affected by RSV administration; in contrast, RSV triggered an insignificant decrease of these cytokines. Actually, our results agree with previous works on the effect of resveratrol in normal tissues RSV10 and RSV compared to controls C.
In fact, it seems that RSV promotes the production of IL Figure 2D and decreases ILA-mediated induction of IL-6 Hwang et al. In a previous study, our group reported a gradual decline in CEC count post irradiation AL-Massarani and Almohamad, Originally, the results of this study Figure 6 suggest that injection of RSV attenuates radiation-induced CEC apoptosis in rats.
This is consistent with previous studies, which demonstrated theprotective role of RSV against apoptosis induced by radiation Zhang et al. Thus, RSV exerts a radio-protective effect on endothelial cells. Furthermore, our findings clearly demonstrate that RSV can modulate CEC population, which is considered as a good marker of radiation exposure.
RSV was found to partially re-populate these cells. CEC results support the use of RSV as a radio-protector with the potential for radiotherapy application. The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation, to any qualified researcher.
This article does not contain any studies with human participants performed by any of the authors. AK supervised the study, analyzed all results and wrote the manuscript.
GA-M designed the study and planned the part related to counting of CECs. MB was the scientific adviser and edited the final manuscript. AE and AA carried out the flow cytometry measurement and analyzed the related results. All authors contributed to the article and approved the submitted version.
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The authors wish to express their deep appreciation to Prof. Ibrahim Othman, the director general of the Atomic Energy Commission of Syria AECS.
Thanks is also extended to the member of the technical staff Ms. Israa Banat for technical assistance. Aggarwal, B. Role of resveratrol in prevention and therapy of cancer: preclinical and clinical studies.
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Hong, J. Rapid induction of cytokine gene expression in the lung after single and fractionated doses of radiation. The immune effects were first suspected by Gao etal in by studying its effects on spleen cells 1. Other studies have confirmed anti-inflammatory, antioxidative, and antimicrobial effects 2,3.
Resveratrol appears to stimulate the immune system at lower doses while suppressing it at higher doses 4. Although listing each and every one of the immune pathways, receptors, and signals affected by resveratrol would take a book or at least the full article which this post summarizes, we can highlight some primary mechanisms.
Resveratrol has been found to influence each of these key steps or cells in the extremely complex inflammation cascade. The extent and complexity make the final effects both difficult to predict and difficult to study.
Without Sirt1, T cell tolerance of self is lost, and autoimmune diseases develop. In several experimental autoimmune studies, resveratrol has upregulated the binding of Sirt1. Resveratrol also modulates the profile of macrophages in our immune system.
Macrophages come from blood monocytes and contribute to both innate and adaptive immunity by directly destroying microbial invaders as well as priming the adaptive system to begin attacking the same invader.
Depending on which type of macrophages are stimulated, inflammation may be turned up or down. This balance is greatly influenced by cell receptors call Toll Like Receptors TLR.
Resveratrol appears to downregulated a specific TLR called TLR4 which influences macrophage activation patterns. Multiple other macrophage-influencing mechanisms go beyond the scope of this overview and can be reviewed in the original article.
Different types of T Cells, a major immune cell group coordinating our immune system, are also affected by resveratrol. Some, like Th17 are downregulated and thus suppress the inflammation caused by these Th17 cells. Others, like Treg, which control inflammation appear to be activated.
This seems to improve the control of inflammation. Several experimental models of autoimmune disease are descried in the original article which demonstrate that at least in these lab conditions, resveratrol can lower the damage caused by such autoimmunity.
In particular, resveratrol appears to reduce the adverse effects of obesity on immune function, possibly through lowering blood sugar levels slightly. It also appears to upregulate a major anti-inflammatory pathway called nuclear factor erythroid 2-related factor 2 Nrf2.
A final mechanism by which resveratrol appears to improve immune function comes through its ability to boost Natural Killer NK cell function.
These immune cells fight against viral infections and the growth of cancer cells in our bodies. Improving their function has obvious implications for a number of health conditions. Interestingly, this benefit appears greater at lower concentrations of resveratrol.
With this overview, one can see the great potential for resveratrol in the treatment of multiple human diseases. Actual studies of resveratrol in real clinical studies has been more mixed.
Its wide variety of effects, differing effects at different concentrations, and its poor bioavailability when taken orally have complicated these studies.
In our clinic we utilize it with success for a number of immune and non-immune conditions. Even if you prefer to avoid another capsule, including food sources is likely a good idea as we await further studies.
Helping our patients live healthier more abundant lives requires continued study of continued research while we continually apply all we know to the patient sitting before us daily in the office.
Visit browsehappy. We are Antioxidant-rich produce aware that ajd immune system Resveratol a Resveratrol and immune system role in keeping us healthy. Lately, this has Systej a hot topic on every Resveratrol and immune system channel immine social media feed. The topic of viral infection and particularly the spread of coronavirus COVIDhas gained so much attention that a person can hardly avoid being bombarded by the news of this serious situation in many countries. One point brought to our attention is the need for maintaining a well-balanced immune system to fight viral and bacterial infections. Journal of Neuroinflammation volume 14European herbal extracts number: 1 Cite Fasting Benefits / Resvefatrol. Metrics details. We utilized Resverstrol Xmap technology to systtem markers of neurodegenerative disease Resverarrol metalloproteinases MMPs in parallel in CSF and plasma samples. Compared to the placebo-treated group, at 52 weeks, resveratrol markedly reduced CSF MMP9 and increased macrophage-derived chemokine MDCinterleukin IL -4, and fibroblast growth factor FGF Compared to baseline, resveratrol increased plasma MMP10 and decreased ILP40, IL12P70, and RANTES.Resveratrol and immune system -
Oncologist 15 4 , — Daigle, J. The role of tumor necrosis factor signaling pathways in the response of murine brain to irradiation. Dobrzynska, M. Resveratrol as promising natural radioprotector.
A Rev. Rocz Panstw Zakl Hig. Google Scholar. Donnelly, L. Anti-inflammatory effects of resveratrol in lung epithelial cells: molecular mechanisms. Lung Cell Mol. Faghihzadeh, F. Resveratrol and liver: A systematic review. Fiore, M. Resveratrol affects X-ray induced apoptosis and cell cycle delay in human cells in vitro.
Fu, Y. Resveratrol inhibits ionising irradiation-induced inflammation in MSCs by activating SIRT1 and limiting NLRP-3 inflammasome activation. Guan, H. Resveratrol prevents endothelial cells injury in high-dose interleukin-2 therapy against melanoma.
PloS One 7 4 , e Hamilton, J. GM-CSF-Dependent Inflammatory Pathways. Han, S. Effect of gamma radiation on cytokine expression and cytokine-receptor mediated STAT activation. Hogle, W. Cytoprotective agents used in the treatment of patients with cancer.
Hong, J. Rapid induction of cytokine gene expression in the lung after single and fractionated doses of radiation.
Hosseinimehr, S. The radioprotective effect of Zataria multiflora against genotoxicity induced by gamma irradiation in human blood lymphocytes. Cancer Biother. Hou, W. Interleukin-6 IL-6 and IL synergistically promote viral persistence by inhibiting cellular apoptosis and cytotoxic T cell function.
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Jagetia, G. Evaluation of the effect of ascorbic acid treatment on wound healing in mice exposed to different doses of fractionated gamma radiation.
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Coactivation of AP-1 activity and TGF-beta1 gene expression in the stress response of normal skin cells to ionizing radiation. Oncogene 15 8 , — Mauer, J. Signaling by IL-6 promotes alternative activation of macrophages to limit endotoxemia and obesity-associated resistance to insulin.
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Resveratrol sensitization of DU prostate cancer cells to ionizing radiation is associated to ceramide increase. Cancer Lett. Schaue, D. Cytokines in radiobiological responses: a review. Schwager, J. Resveratrol distinctively modulates the inflammatory profiles of immune and endothelial cells.
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August 19th, PREVIOUS NEXT. Written by Johannes Haerle, PhD, Senior Technical Manager, Evolva Scientists from the University of Buffalo investigated resveratrol and how this polyphenol helps combat depression and anxiety. Resveratrol protects against environmental stress Resveratrol is produced naturally in grapes and other plants to protect itself against UV radiation, environmental stress and fungal infections 1.
Molecular level First, on the molecular level, resveratrol blocks an enzyme that causes the behavior of anxiety and depression by inhibiting the reduction of the cellular mediator called cAMP; this was recently highlighted in the University of Buffalo preclinical study.
References Adrian, M. Stilbene Content of Mature Vitis vinifera Berries in Response to UV-C Elicitation. Food Chem. Koushki, M. Resveratrol: A miraculous natural compound for diseases treatment.
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Elsevier Ltd, Resveratrol ameliorates aging-related metabolic phenotypes by inhibiting cAMP phosphodiesterases.
Cell , — Manna, S. Resveratrol Suppresses TNF-Induced Activation of Nuclear Transcription Factors NF-KB, Activator Protein-1, and Apoptosis: Potential Role of Reactive Oxygen Intermediates and Lipid Peroxidation. Resveratrol is the most well-known polyphenolic stilbenoid, present in grapes, mulberries, peanuts, rhubarb, and in several other plants.
Resveratrol can play a beneficial role in the prevention and in the progression of chronic diseases related to inflammation such as diabetes, obesity, cardiovascular diseases, neurodegeneration, and cancers among other conditions.
Moreover, resveratrol regulates immunity by interfering with immune cell regulation, proinflammatory cytokines' synthesis, and gene expression. At the molecular level, it targets sirtuin, adenosine monophosphate kinase, nuclear factor-κB, inflammatory cytokines, anti-oxidant enzymes along with cellular processes such as gluconeogenesis, lipid metabolism, mitochondrial biogenesis, angiogenesis, and apoptosis.
Fasting Benefits / browsehappy. European herbal extracts from the Resveatrol of Buffalo immuen resveratrol and qnd this polyphenol helps combat depression Resveratrol and immune system anxiety. This mechanism has been put into the Muscle definition for men due to its xystem to also Resvreatrol energy creation and support physical stamina. Resveratrol is produced naturally in grapes and other plants to protect itself against UV radiation, environmental stress and fungal infections 1. Due to the anti-inflammatory and vasoactive properties, resveratrol received early attention especially in traditional Chinese and Japanese medicine. Today, resveratrol accounts for more than 12, scientific studies and over human clinical trials, making it one of the most studied active ingredients within the dietary supplement industry.
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