Abstract 3310: Human Peripheral Blood-derived CD31 + Cells Have Robust Angio-vasculogenic Properties And Enhance Recovery Following Hindlimb Ischemia

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Sung-Whan Kim ◽  
Hyun-Jai Cho ◽  
Ji Yoon Lee ◽  
Yong Jin Choi ◽  
Young-sup Yoon
Keyword(s):  
Peripheral Blood ◽  
Network Formation ◽  
Therapeutic Potential ◽  
Human Peripheral Blood ◽  
Therapeutic Option ◽  
Fold Increase ◽  
Treated Group ◽  
Circulating Endothelial Cells ◽  
Hindlimb Ischemia ◽  
Murine Bone Marrow

Background: Recently, we reported that murine bone marrow derived CD31 + cells have hemangioblastic activities. In this study, we hypothesized that human peripheral blood derived CD31 + cells (hCD31 + ) have robust angio-vasculogenic potential and can induce therapeutic neovascularization in hindlimb ischemia. Methods and Results: hCD31 + cells were isolated using a magnetic bead separation technique. By FACS analysis, we confirmed that more than 98% of hCD31 + cells are positive for CD45, suggesting that hCD31 + cells are not circulating endothelial cells. In endothelial progenitor cell (EPC) culture assay, hCD31 + cells almost exclusively gave rise to EPCs and produced significantly higher colony forming activity than hCD31 - cells (n=5, each) (25.2 ± 2.2 vs. 3.6 ± 0.5/mm 2 , P<0.01). In the matrigel tube formation assay, coculturing HUVEC with hCD31 + cells revealed that hCD31 + cells robustly contributed to network formation with HUVECs suggestive of angio-vasculogenic activity. Real-time PCR showed that hCD31 + cells express higher levels of angiogenic genes (fold increase: angiopoietin-1, 5.6 ± 0.8; HGF, 2.7 ± 0.6; VEGF-A, 3.8 ± 0.5; FGF-2, 2.0 ± 0.5) and lower levels of inflammatory genes (fold decrease: Interferonγ, 13.2 ± 2.5; TNFα, 2.3 ± 0.9) than hCD31 − cells. To investigate therapeutic potential, we surgically induced hindlimb ischemia in athymic nude mice, and injected hCD31 + cells, hCD31 − cells and PBS into the ischemic limbs (n=9, each). The hCD31 + treated group showed a higher limb salvage rate than other groups [total salvage/tip necrosis/amputation; hCD31 + cells 5/4/0, hCD31 − cells 2/5/2, PBS control 0/5/4] (P<0.01). At 14 days, perfusion ratio and capillary density were significantly higher in the hCD31 + treated group compared to the hCD31 − and PBS treated groups. Histologic analysis demonstrated that a fraction of injected DiI-labeled hCD31 + cells exhibit endothelial phenotypes during the follow-up period of 8 weeks. Conclusion: The present study demonstrated that hCD31 + cells have robust angio-vasculogenic potential with low inflammatory activity, and enhanced post-ischemia recovery. These data suggested that hCD31 + cell transplantation may represent a novel therapeutic option for treating ischemic cardiovascular diseases.

scholarly journals Liposome/gold hybrid nanoparticle encoded with CoQ10 (LGNP-CoQ10) suppressed rheumatoid arthritis via STAT3/Th17 targeting

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0241080
Author(s):  
Jooyeon Jhun ◽  
Jeonghyeon Moon ◽  
Jaeyoon Ryu ◽  
Yonghee Shin ◽  
Seangyoun Lee ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Mononuclear Cells ◽  
Therapeutic Potential ◽  
Human Peripheral Blood ◽  
Treated Group ◽  
Hybrid Nanoparticles ◽  
Peripheral Blood Mononuclear ◽  
Hematoxylin And Eosin ◽  
Anti Inflammatory ◽  
Safranin O

Coenzyme Q10 (CoQ10), also known as ubiquinone, is a fat-soluble antioxidant. Although CoQ10 has not been approved as medication by the Food and Drug Administration, it is widely used in dietary supplements. Some studies have shown that CoQ10 has anti-inflammatory effects on various autoimmune disorders. In this study, we investigated the anti-inflammatory effects of liposome/gold hybrid nanoparticles encoded with CoQ10 (LGNP-CoQ10). Both CoQ10 and LGNP-CoQ10 were administered orally to mice with collagen-induced arthritis (CIA) for 10 weeks. The inflammation pathology of joint tissues of CIA mice was then analyzed using hematoxylin and eosin and Safranin O staining, as well as immunohistochemistry analysis. We obtained immunofluorescence staining images of spleen tissues using confocal microscopy. We found that pro-inflammatory cytokines were significantly decreased in LGNP-CoQ10 injected mice. Th17 cell and phosphorylated STAT3-expressed cell populations were also decreased in LGNP-CoQ10 injected mice. When human peripheral blood mononuclear cells (PBMCs) were treated with CoQ10 and LGNP-CoQ10, the IL-17 expression of PBMCs in the LGNP-CoQ10-treated group was significantly reduced. Together, these results suggest that LGNP-CoQ10 has therapeutic potential for the treatment of rheumatoid arthritis.

The therapeutic potential of ex vivo expanded CD133+ cells derived from human peripheral blood for peripheral nerve injuries

2012 ◽  
Vol 117 (4) ◽  
pp. 787-794 ◽  
Author(s):  
Shin Ohtsubo ◽  
Masakazu Ishikawa ◽  
Naosuke Kamei ◽  
Yasumu Kijima ◽  
Osami Suzuki ◽  
...  
Keyword(s):  
Sciatic Nerve ◽  
Peripheral Nerve ◽  
Peripheral Blood ◽  
Therapeutic Potential ◽  
Ex Vivo ◽  
Human Peripheral Blood ◽  
Peripheral Nerve Injuries ◽  
Nerve Injuries ◽  
Group 4 ◽  
Group 1

Object CD133+ cells have the potential to enhance histological and functional recovery from peripheral nerve injury. However, the number of CD133+ cells safely obtained from human peripheral blood is extremely limited. To address this issue, the authors expanded CD133+ cells derived from human peripheral blood using the serum-free expansion culture method and transplanted these ex vivo expanded cells into a model of sciatic nerve defect in rats. The purpose of this study was to determine the potential of ex vivo expanded CD133+ cells to induce or enhance the repair of injured peripheral nerves. Methods Phosphate-buffered saline (PBS group [Group 1]), 105 fresh CD133+ cells (fresh group [Group 2]), 105 ex vivo expanded CD133+ cells (expansion group [Group 3]), or 104 fresh CD133+ cells (low-dose group [Group 4]) embedded in atelocollagen gel were transplanted into a silicone tube that was then used to bridge a 15-mm defect in the sciatic nerve of athymic rats (10 animals per group). At 8 weeks postsurgery, histological and functional evaluations of the regenerated tissues were performed. Results After 1 week of expansion culture, the number of cells increased 9.6 ± 3.3–fold. Based on the fluorescence-activated cell sorting analysis, it was demonstrated that the initial freshly isolated CD133+ cell population contained 93.22% ± 0.30% CD133+ cells and further confirmed that the expanded cells had a purity of 59.02% ± 1.58% CD133+ cells. However, the histologically and functionally regenerated nerves bridging the defects were recognized in all rats in Groups 2 and 3 and in 6 of 10 rats in Group 4. The nerves did not regenerate to bridge the defect in any of the rats in Group 1. Conclusions The authors' results show that ex vivo expanded CD133+ cells derived from human peripheral blood have a therapeutic potential similar to fresh CD133+ cells for peripheral nerve injuries. The ex vivo procedure that can be used to expand CD133+ cells without reducing their function represents a novel method for developing cell therapy for nerve defects in a clinical setting.

Selective mobilization of CD14+CD16+monocytes by exercise

2000 ◽  
Vol 279 (3) ◽  
pp. C578-C586 ◽  
Author(s):  
Birgit Steppich ◽  
Farshid Dayyani ◽  
Rudolf Gruber ◽  
Reinhard Lorenz ◽  
Matthias Mack ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Peripheral Blood ◽  
Adrenergic Receptor ◽  
Initial Level ◽  
Human Peripheral Blood ◽  
Fold Increase ◽  
Anaerobic Exercise ◽  
Late Antigen ◽  
Clinical Conditions ◽  
Dependent Mechanism

Strenuous, anaerobic exercise leads to an increase of leukocytes that are mobilized from the marginal pool. We have analyzed in human peripheral blood the effect of exercise on the number of CD14+CD16+monocytes as determined by two-color immunofluorescence and flow cytometry. We show herein that this type of monocyte responds with a dramatic up to 4.8-fold increase. Mobilization does not occur after 1 min at 100 or 200 W but 1 min at 400 W leads to a twofold increase of the CD14+CD16+monocytes immediately after exercise. The numbers remain high at 5 min and gradually decrease to reach the initial level at 20 min postexercise. After 20 min of rest, the CD14+CD16+monocytes can be mobilized again by a second exercise. The CD14+CD16+monocytes appear to be mobilized from the marginal pool where they preferentially home because of a higher expression of adhesion molecules like CD11d and very late antigen-4. Exercise goes along with an increase of catecholamines, and mobilization of the CD14+CD16+monocytes can be substantially reduced by treatment of donors with the β-adrenergic receptor blocker propranolol. Mobilization of CD14+CD16+monocytes by a catecholamine-dependent mechanism may contribute to the increase of these cells in various clinical conditions.

Abstract 17581: Evidence for Circulating Stem Cells Derived from the Human Heart and Their Therapeutic Potential

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Han-Mo Yang ◽  
Ju-Young Kim ◽  
Hyun-Jai Cho ◽  
Joo-Eun Lee ◽  
So-Jung Lim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Peripheral Blood ◽  
Human Heart ◽  
Therapeutic Potential ◽  
Kidney Diseases ◽  
Heart Function ◽  
Human Peripheral Blood ◽  
Mouse Heart ◽  
Multipotent Stem Cells ◽  
Hematopoietic Stem

Introduction and Hypothesis: Many studies have shown resident cardiac stem cells in myocardium as well as epicardial progenitor cells in epicardium. However, the presence of stem cells in the endocardium has not been elucidated. In this study, we identified circulating multipotent stem cells from human peripheral blood. Furthermore, we investigated the origin and the therapeutic potential of these cells. Methods and Results: We identified a new population of cells from human peripheral blood mononuclear cells, which were quite different from previously reported stem cells. Newly identified cells expressed genes such as Oct3/4, KLF4, Nanog, and c-Myc. Moreover, FACS analysis precluded the possibility that these cells might be hematopoietic stem cells. To investigate the origin of these cells, we collected peripheral blood from patients undergoing bone marrow, liver, heart, or kidney transplantation. After culturing these cells, we could confirm that these stem cells were derived from the human heart by identifying the HLA types or the STR (short tandem repeat) profiles. In addition, we demonstrated that Nuclear Factor of Activated T-cells (NFAT)-positive and CD31-positive circulating cells in peripheral blood were derived from NFAT-positive cells in the endocardium. These cells had multipotency, indicating the ability of differentiation not only into mesodermal lineages, but also into ectodermal or endodermal lineages. When injected into the mouse heart in vivo , these stem cells were differentiated into multiple lineages, resulting in the improvement of the heart function. We established more than 200 cell lines from peripheral blood of patients with coronary artery diseases, cardiomyopathies, hematologic diseases, liver diseases, and kidney diseases. Conclusions: We demonstrated the existence of novel circulating multipotent stem cells in human peripheral blood, which express NFAT. Interestingly, these cells are derived from the tissue-resident stem cells of the endocardium of the human heart. Our findings suggest that these stem cells obtainable from peripheral blood could be a promising tool for heart regeneration.

AMD3100 and G-CSF Mobilize Angiogenic Cells into the Blood of Healthy Donors.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3022-3022
Author(s):  
Rebecca M. Shepherd ◽  
Benjamin J. Capoccia ◽  
Steven M. Devine ◽  
David A. Ingram ◽  
Daniel C. Link
Keyword(s):  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Fold Increase ◽  
Peripheral Circulation ◽  
Adherent Cells ◽  
Hindlimb Ischemia ◽  
Angiogenic Potential ◽  
Ischemic Limb ◽  
Limb Perfusion

Abstract There is evidence that angiogenic cells exist in the blood that are able to home to sites of ischemia and induce angiogenesis. Angiogenic cell populations identified in human blood include endothelial progenitor cells (EPC) and circulating angiogenic monocytes (CAM). Under basal conditions, however, the number of angiogenic cells in the circulation is small, potentially limiting their delivery to sites of ischemia and subsequent stimulation of angiogenesis. To circumvent this limitation, animal studies have shown that angiogenic cells can be mobilized into blood by certain cytokines; whether a similar mobilization of angiogenic cells can be achieved in humans is not clear. Herein, we report the results of a clinical trial that examined the ability of G-CSF and AMD3100 to mobilize EPC and CAM into the blood. Donors were treated with a single injection of AMD3100 (5mg/kg) after which pheresis was performed. One week later, donors were treated with G-CSF (250microg/kg/d x 5 days) and pheresed. Blood was collected at baseline (prior to the initiation of the mobilizing regimen) and after treatment with AMD3100 or G-CSF (at the time of peak HSC mobilization). In addition, pheresis product was collected after mobilization by AMD3100 or G-CSF. Mononuclear cells (MNC) were isolated from the blood products and cultured under angiogenic conditions. EPC were identified by the formation of discrete colonies of endothelial cells on days 14-28. A novel method to quantify CAM was developed to avoid the pitfalls of counting adherent cells in culture. Briefly, adherent cells were recovered from the culture, counted, and analyzed by flow cytometry. CAM were identified as CD45+ CD14+ CD31+ cells. The angiogenic potential of the CAM was assessed by their transplantation into NOD/SCID/b2M−/− mice following surgical induction of hindlimb ischemia. Indeed, reperfusion was significantly enhanced in mice that received CAM compared to control mice [ratio±SEM) of ischemic to non-ischemic limb perfusion: 0.47±0.05 (saline treated); 0.78±0.09 (CAM transplant); p<.001]. To date, a total of 9 patients have been analyzed. AMD3100 resulted in a 9.2-fold increase in CAM in the blood [CAM/ml peripheral blood±SEM: 3.5x105±0.09 (AMD3100) vs. 0.4x105±0.01 (baseline); p<.05]. Likewise, treatment with G-CSF resulted in a 12.3-fold increase in CAM (4.5x105±0.21; p<.05). Both AMD3100 and G-CSF also mobilized EPC into the blood [EPC/mL peripheral blood±SEM: 0.53±0.22 (AMD3100); 0.98±0.24 (G-CSF); vs. 0.05±0.01 (baseline); p<.05 for both AMD3100 and G-CSF compared with baseline]. Interestingly, data suggests that the G-CSF-mobilized CAM may have an enhanced capacity to stimulate angiogenesis in the hindlimb ischemia model [the ratio of ischemic to non-ischemic limb perfusion±SEM: 0.47±0.05 (saline treated); 0.64+0.11 (G-CSF) p<.001; 0.58±0.06 (AMD3100); p=NS]. Importantly, CAM and EPC were readily recovered from the pheresis product after both AMD3100 and G-CSF treatment. Collectively these data show that treatment with AMD3100 or G-CSF mobilizes a significant number of CAM and EPC into the peripheral circulation. These mobilized angiogenic cells have the capacity to stimulate angiogenesis in vivo. Finally, CAM and EPC can be efficiently harvested by leukopheresis, providing a method to obtain large numbers of circulating angiogenic cells for subsequent clinical use.±

HSP27 modulates survival signaling in endosulfan-exposed human peripheral blood mononuclear cells treated with curcumin

2015 ◽  
Vol 35 (7) ◽  
pp. 695-704 ◽  
Author(s):  
T Ahmed ◽  
BD Banerjee
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Therapeutic Potential ◽  
Cell Damage ◽  
Human Peripheral Blood ◽  
Cellular Level ◽  
Peripheral Blood Mononuclear ◽  
Human Pbmc ◽  
Blood Mononuclear Cells

Endosulfan, a well-known organochlorine pesticide, induces apoptosis and depletion of reduced glutathione (GSH) in human peripheral blood mononuclear cells (PBMC). Thus, for the amelioration of its effect, antioxidant and antiapoptotic potential of curcumin was evaluated. For ascertaining the attenuating effect of curcumin, various biochemical indices of cell damage such as cytotoxicity, oxidative stress, apoptosis (phosphatidylserine externalization, DNA fragmentation, and cytochrome c) in human PBMC was evaluated following endosulfan exposure (0–100 µM). To assess the role of HSP27 on endosulfan-induced apoptosis, the expression of HSP27 was examined. Curcumin (25 µM) increased cell viability significantly. As evident from the restoration of GSH, antiapoptotic potential was directly proportional to their antioxidant nature of curcumin. The present study indicates that the beneficial effect of curcumin on endosulfan-induced cytotoxicity is related to the induced synthesis of HSP27, emphasizing its antioxidant and therapeutic potential as well as underscoring the mechanism of pesticide-induced toxicity at cellular level. Taken together, these findings suggest that curcumin protects against endosulfan-induced immunotoxicity in human PBMC by attenuating apoptosis.

scholarly journals Ex VivoNicotine Stimulation Augments the Efficacy of Human Peripheral Blood Mononuclear Cell-Derived Dendritic Cell Vaccination via Activating Akt-S6 Pathway

2015 ◽  
Vol 2015 ◽  
pp. 1-13 ◽  
Author(s):  
Yan Yan Wang ◽  
Yi Wen Yang ◽  
Xiang You ◽  
Xiao Qian Deng ◽  
Chun Fang Hu ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cell ◽  
Mononuclear Cell ◽  
Peripheral Blood ◽  
Ex Vivo ◽  
Human Peripheral Blood ◽  
Dendritic Cell Vaccination ◽  
Peripheral Blood Mononuclear ◽  
Murine Bone Marrow ◽  
Cross Presentation ◽  
Surface Molecules

Our previous studies showed thatα7 nicotinic acetylcholine receptor (nAchR) agonist nicotine has stimulatory effects on murine bone marrow-derived semimature DCs, but the effect of nicotine on peripheral blood mononuclear cell- (PBMC-) derived human semimature dendritic cells (hu-imDCs) is still to be clarified. In the present study, hu-imDCs (cultured 4 days) were conferred with  ex vivolower dose nicotine stimulation and the effect of nicotine on surface molecules expression, the ability of cross-presentation, DCs-mediated PBMC priming, and activated signaling pathways were determined. We could demonstrate that the treatment with nicotine resulted in increased surface molecules expression, enhanced hu-imDCs-mediated PBMC proliferation, upregulated release of IL-12 in the supernatant of cocultured DCs-PBMC, and augmented phosphorylation of Akt and ribosomal protein S6. Nicotine associated with traces of LPS efficiently enhanced endosomal translocation of internalized ovalbumin (OVA) and increased TAP-OVA colocalization. Importantly, the upregulation of nicotine-increased surface molecules upregulation was significantly abrogated by the inhibition of Akt kinase. These findings demonstrate thatex vivonicotine stimulation augments hu-imDCs surface molecules expression via Akt-S6 pathway, combined with increased Ag-presentation result in augmented efficacy of DCs-mediated PBMC proliferation and Th1 polarization.

Increased Angiogenic Property Of Human Peripheral Blood Monocytes By ex Vivo Culture With c-Mpl Agonists In Hindlimb Ischemia Mouse Model

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1062-1062
Author(s):  
Junpei Sasajima ◽  
Toru Kawamoto ◽  
Yoshiaki Sugiyama ◽  
Yusuke Mizukami ◽  
Yasuyuki Iuchi ◽  
...  
Keyword(s):  
Growth Factor ◽  
Peripheral Blood ◽  
Culture System ◽  
Intramuscular Injection ◽  
Ex Vivo ◽  
Human Peripheral Blood ◽  
Autologous Serum ◽  
Hindlimb Ischemia ◽  
Serum Free ◽  
Ex Vivo Culture

Abstract Human peripheral blood mononuclear cells (PB-MNCs) include some populations which have angiogenic properties. Pro-angiogenic monocytes from PB-MNCs are considered as one of candidates for angiogenic therapy in regenerative medicine. Indeed, in a recent German clinical post-infarction remodeling study (TOPCARE-AMI) for ischemic heart disease, the ex vivo culture of PB-MNCs was employed. However, in this trial, there were different therapeutic efficacies in each case, possibly due to the different expansion efficacy of the ex vivo culture of PB-MNCs using autologous serum. In order to resolve this issue, we developed a new serum-free culture system composed of X-VIVO15 medium supplemented with vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Floating spheres obtained by this serum-free culture system were mainly composed of CD11b+ monocytes. Interestingly, the mRNA expression of c-Mpl (thrombopoietin receptor) was markedly elevated compared with PB-MNCs, suggesting c-Mpl agonist could increase angiogenic property of cultured CD11b+ monocytes. Therefore, we assessed the impact of c-Mpl agonists on PB-MNC cultures in our serum-free method composed of X-VIVO15 medium with VEGF and bFGF. Both recombinant human thrombopoietin (rHuTPO) and romiplostim, a clinical grade second-generation TPO-receptor agonist, successfully increased sphere formations regarding both the number and size. The expressions of angiogenic factors, IL-8, CXCR4, and vasohibin-2, mRNA of CD11b+ monocytes cultured with c-Mpl agonists were up-regulated, indicating that cultivated CD11b+ monocytes have a proangiogenic potential. Finally, we investigated the proangiogenic potential of PB-MNCs derived CD11b+ monocytes in a hindlimb ischemia model utilizing BALB/c nude mice. Mice were randomly assigned to 7 groups: control mouse group (PBS-injected), freshly isolated CD11b+ monocyte-injected mouse group, cultivated CD11b+ monocyte with 2ng/ml and 20ng/ml rHuTPO -injected mouse groups, cultivated CD11b+ monocyte with 100ng/ml and 1000ng/ml romiplostim -injected mouse groups, cultivated CD11b+ monocyte without rHuTPO and romiplostim -injected mouse group. The intramuscular injection of CD11b+ monocytes cultivated with 20 ng/ml rHuTPO into the ischemic limb completely rescued the limbs from auto-amputations or foot necrosis, while only one (10.0%) of the control mice could be rescued. In addition, the intramuscular injection of both freshly isolated CD11b+ monocytes and CD11b+ monocytes cultivated without rHuTPO and romiplostim had a weak rescue effect on the ischemic limbs (8 and 7 of 10 mice had auto-amputations or foot necrosis, respectively). The salvage rate from necrosis in cultivated CD11b+ monocyte with romiplostim-injected mouse group is also superior to that in cultivated CD11b+ monocyte without rHuTPO-injected mouse group. Analysis of blood perfusion by a laser Doppler perfusion imaging system showed a significantly higher recovery in mice receiving the CD11b+ monocytes cultivated with 2 ng/ml or 20 ng/ml rHuTPO or 100ng/ml romiplostim 1 week after surgery. The functional capillary density and surface area visualized by perfusion with BS-I lectin also significantly increased in the rHuTPO- or romiplostim-treated group. In conclusion, an ex vivo addition of c-Mpl agonists augmented the pro-angiogenic activity of peripheral CD11b+ monocytes, and this method would be promising for human cell therapy to induce vascular regeneration by activating the angiogenic property in human peripheral blood-derived monocytes. Disclosures: Mizukami: The New Energy and Industrial Technology Development Organization of Japan: Research Funding.

scholarly journals Can Melatonin Act as an Antioxidant in Hydrogen Peroxide-Induced Oxidative Stress Model in Human Peripheral Blood Mononuclear Cells?

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Solaleh Emamgholipour ◽  
Arash Hossein-Nezhad ◽  
Mohammad Ansari
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Fold Increase ◽  
Gene Expressions ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Purpose.We aimed to investigate the possible effects of melatonin on gene expressions and activities of MnSOD and catalase under conditions of oxidative stress induced by hydrogen peroxide (H2O2) in peripheral blood mononuclear cells (PBMCs).Materials and Methods.PBMCs were isolated from healthy subjects and treated as follows: (1) control (only with 0.1% DMSO for 12 h); (2) melatonin (1 mM) for 12 h; (3) H2O2(250 μM) for 2 h; (4) H2O2(250 μM) for 2 h following 10 h pretreatment with melatonin (1 mM). The gene expression was evaluated by real-time PCR. MnSOD and catalase activities in PBMCs were determined by colorimetric assays.Results.Pretreatment of PBMCs with melatonin significantly augmented expression and activity of MnSOD which were diminished by H2O2. Melatonin treatment of PBMCs caused a significant upregulation of catalase by almost 2-fold in comparison with untreated cells. However, activity and expression of catalase increased by 1.5-fold in PBMCs under H2O2-induced oxidative stress compared with untreated cell. Moreover, pretreatment of PBMCs with melatonin resulted in a significant 1.8-fold increase in catalase expression compared to PBMCs treated only with H2O2.Conclusion.It seems that melatonin could prevent from undesirable impacts of H2O2-induced oxidative stress on MnSOD downregulation. Moreover, melatonin could promote inductive effect of H2O2on catalase mRNA expression.

Sign in / Sign up

Export Citation Format

Share Document