scholarly journals Effect of grape seed proanthocyanidin extract in peripheral blood mononuclear cells of severe asthmatic patients

2019 ◽  
Author(s):  
Yun Sun ◽  
Bin Gu ◽  
Yan Qian ◽  
Yi Chen ◽  
Zheng-dao Mao ◽  
...  
Keyword(s):  
Severe Asthma ◽  
Mononuclear Cells ◽  
Grape Seed ◽  
Control Group ◽  
Binding Ability ◽  
Peripheral Blood Mononuclear ◽  
Asthmatic Patients ◽  
Severe Asthmatic ◽  
Blood Mononuclear Cells ◽  
Grape Seed Proanthocyanidin

Abstract Backgroud To explore the Effect of grape seed proanthocyanidin extract（GSPE） in peripheral blood mononuclear cells of severe asthmatic patients. Methods 40 severe asthmatic patients were randomly and averagely divided into 2 groups: DXM group (n=20) and GSPE+DXM group (n=20), and 20 healthy volunteers as control gorup. Heparinized peripheral venous blood from each subject was collected in a sterile vacuum tube. The levels of interleukin (IL)-8 and monocyte chemotactic protein (MCP)-1 were detected by ELISA. The protein and mRNA expressions of Nuclear factor erythroid 2-related factor 2 (Nrf2), glutamatecysteine ligase modifier subunit (GCLM) inducible nitric oxide synthase (iNOS) and inactivation of histone deacetylase-2 (HDAC2) were detected by Western blot and qRT-PCR respectively. Glutathion(GSH) was measured by using Glutathione Fluorometric Detection Kit, and Nrf2-ARE binding ability was measured by using TransAM Nrf2 Transcription Factor ELISA Kit. Results The results showed that the levels of IL-8 and MCP-1 in normal control group were lower than those in severe asthma group (P <0.05). Treatment with GSPE+DXM reduced the levels of IL-8 and MCP-1 significantly when compared with DXM only ( P <0.05).The mRNA expression of iNOS in DXM group was significantly higher than that in control group. However, after adding GSPE treatment, the expression dramatically decreased ( P <0.001). On the contrary, lower mRNA expressions of Nrf2, GCLM and HDAC2 were found in DXM group than in control group ( P <0.001). Accordingly, when treated with GSPE, these expressions elevated and reached a statistical significance ( P <0.05). Consistently, the results from western blot analysis confirmed the role of GSPE on the protein expressions of iNOS, Nrf2, GCLM and HDAC2 in PBMCs of patients with severe asthma ( P <0.001). The PBMCs from patients with severe asthma exhibited lower Nrf2-ARE binding ability and produced less GSH than normal controls. GSPE treatment effectively augmented the Nrf2-ARE binding ability and the expression of GSH, which demonstrated the biological antioxidant potential of GSPE ( P <0.001). Conclusion GSPE can alleviate glucocorticoid resistance by regulating Nrf2-iNOS-HDAC2 signaling pathway in severe asthma, suggesting that GSPE have potential clinical application prospects in the treatment of severe asthma.

scholarly journals Effect of grape seed proanthocyanidin extract in peripheral blood mononuclear cells of severe asthmatic patients

2020 ◽  
Author(s):  
Yun Sun ◽  
Bin Gu ◽  
Yan Qian ◽  
Yi Chen ◽  
Zheng-dao Mao ◽  
...  
Keyword(s):  
Severe Asthma ◽  
Mononuclear Cells ◽  
Grape Seed ◽  
Control Group ◽  
Binding Ability ◽  
Peripheral Blood Mononuclear ◽  
Asthmatic Patients ◽  
Severe Asthmatic ◽  
Blood Mononuclear Cells ◽  
Grape Seed Proanthocyanidin

Abstract Backgroud To explore the Effect of grape seed proanthocyanidin extract（GSPE） in peripheral blood mononuclear cells of severe asthmatic patients. Methods 40 severe asthmatic patients were randomly and averagely divided into 2 groups: DXM group (n=20) and GSPE+DXM group (n=20), and 20 healthy volunteers as control gorup. Heparinized peripheral venous blood from each subject was collected in a sterile vacuum tube. The levels of interleukin (IL)-8 and monocyte chemotactic protein (MCP)-1 were detected by ELISA. The protein and mRNA expressions of Nuclear factor erythroid 2-related factor 2 (Nrf2), glutamatecysteine ligase modifier subunit (GCLM) inducible nitric oxide synthase (iNOS) and inactivation of histone deacetylase-2 (HDAC2) were detected by Western blot and qRT-PCR respectively. Glutathion(GSH) was measured by using Glutathione Fluorometric Detection Kit, and Nrf2-ARE binding ability was measured by using TransAM Nrf2 Transcription Factor ELISA Kit. Results The results showed that the levels of IL-8 and MCP-1 in normal control group were lower than those in severe asthma group (P <0.05). Treatment with GSPE+DXM reduced the levels of IL-8 and MCP-1 significantly when compared with DXM only ( P <0.05).The mRNA expression of iNOS in DXM group was significantly higher than that in control group. However, after adding GSPE treatment, the expression dramatically decreased ( P <0.001). On the contrary, lower mRNA expressions of Nrf2, GCLM and HDAC2 were found in DXM group than in control group ( P <0.001). Accordingly, when treated with GSPE, these expressions elevated and reached a statistical significance ( P <0.05). Consistently, the results from western blot analysis confirmed the role of GSPE on the protein expressions of iNOS, Nrf2, GCLM and HDAC2 in PBMCs of patients with severe asthma ( P <0.001). The PBMCs from patients with severe asthma exhibited lower Nrf2-ARE binding ability and produced less GSH than normal controls. GSPE treatment effectively augmented the Nrf2-ARE binding ability and the expression of GSH, which demonstrated the biological antioxidant potential of GSPE ( P <0.001). Conclusion GSPE can alleviate glucocorticoid resistance by regulating Nrf2-iNOS-HDAC2 signaling pathway in severe asthma, suggesting that GSPE have potential clinical application prospects in the treatment of severe asthma.

scholarly journals Mitochondrial Function in Peripheral Blood Mononuclear Cells (PBMC) Is Enhanced, Together with Increased Reactive Oxygen Species, in Severe Asthmatic Patients in Exacerbation

2019 ◽  
Vol 8 (10) ◽  
pp. 1613 ◽  
Author(s):  
Ederlé ◽  
Charles ◽  
Khayath ◽  
Poirot ◽  
Meyer ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Mitochondrial Respiratory Chain ◽  
Ros Production ◽  
Oxygen Species ◽  
Respiratory Chain Complexes ◽  
Peripheral Blood Mononuclear ◽  
Asthmatic Patients ◽  
Chain Complexes ◽  
Severe Asthmatic ◽  
Blood Mononuclear Cells

asthma is a chronic inflammatory lung syndrome with an increasing prevalence and a rare but significant risk of death. Its pathophysiology is complex, and therefore we investigated at the systemic level a potential implication of oxidative stress and of peripheral blood mononuclear cells’ (PBMC) mitochondrial function. Twenty severe asthmatic patients with severe exacerbation (GINA 4–5) and 20 healthy volunteers participated at the study. Mitochondrial respiratory chain complexes activities using different substrates and reactive oxygen species (ROS) production were determined in both groups by high-resolution respirometry and electronic paramagnetic resonance, respectively. Healthy PBMC were also incubated with a pool of plasma of severe asthmatics or healthy controls. Mitochondrial respiratory chain complexes activity (+52.45%, p = 0.015 for VADP) and ROS production (+34.3%, p = 0.02) were increased in asthmatic patients. Increased ROS did not originate mainly from mitochondria. Plasma of severe asthmatics significantly increased healthy PBMC mitochondrial dioxygen consumption (+56.8%, p = 0.031). In conclusion, such asthma endotype, characterized by increased PMBCs mitochondrial oxidative capacity and ROS production likely related to a plasma constituent, may reflect activation of the immune system. Further studies are needed to determine whether increased PBMC mitochondrial respiration might have protective effects, opening thus new therapeutic approaches.

Changes of IL-4 and IFN-g expression in monocytes and T-helper lymphocytes while modeling of systemic juvenile idiopathic arthritis in Wistar rats

2018 ◽  
pp. 61-69
Author(s):  
В.Н. Сахаров ◽  
П.Ф. Литвицкий ◽  
Е.И. Алексеева ◽  
Н.А. Маянский ◽  
Р.Ш. Закиров
Keyword(s):  
Juvenile Idiopathic Arthritis ◽  
Peripheral Blood Mononuclear Cells ◽  
Wistar Rats ◽  
Mononuclear Cells ◽  
Systemic Juvenile Idiopathic Arthritis ◽  
Control Group ◽  
Peripheral Blood Mononuclear ◽  
Treatment Groups ◽  
Blood Mononuclear Cells ◽  
T Helpers

Цель исследования - изучение перепрограммирования мононуклеарных лейкоцитов на модели системного ювенильного идиопатического артрита (сЮИА), воспроизводимой у крыс Wistar с использованием полного адъюванта Фрейнда и липополисахарида. Методика. сЮИА воспроизведен у 6-месячных крыс-самцов Wistar. На 40-е сут. эксперимента животные были разделены на 3 группы: 1-я группа - контроль; 2-я - группа доксициклина; 3-я - группа дексаметазона. Взятие проб крови у животных проводили на нулевые, 41-е и 55-е сут. Мононуклеарные клетки периферической крови выделяли гравиметрически, после чего окрашивали их на маркеры и внутриклеточные цитокины. Дифференцировали моноциты (CD3-CD4+) и Т-хелперы (CD3+CD4+). Анализировали динамику внутриклеточной экспрессии интерлейкина IL-4 (рассматривали как маркер про-М2 фенотипа, так как в случае выделения из клетки ИЛ-4 служит стимулятором М2 поляризации макрофагов) и IFN-g (как маркер про-М1 фенотипа) по данным проточной цитофлуориметрии. Применяли непараметрический статистический тест Mann-Whitney-Wilcoxon в программе R для статистической обработки данных. Результаты и заключение. При моделировании сЮИА выявлено закономерное изменение фенотипа моноцитов. Применение же доксициклина и дексаметазона приводило к более ранней поляризации их по про-М2-пути в отношении моноцитов (на 41-е сут.) в сравнении с контролем. Про-М1 эффект (на 55-е сут., в сравнении с контролем) выявлен также в группах доксициклина и дексаметазона. У животных разных групп обнаружены характерные динамические изменения внутриклеточной экспрессии цитокинов. Важно, что различная направленность поляризации фенотипа при сЮИА и применении препаратов наблюдается не только у моноцитов, но и у Т-хелперов. The study objective was to evaluate targeted reprogramming of peripheral blood mononuclear cells in systemic juvenile idiopathic arthritis (sJIA) modeled in 6-month-old male Wistar rats by co-administration of complete Freund’s adjuvant and lipopolysaccharide. Methods. On day 40 of the experiment, rats were divided into three groups: control, doxycycline, and dexamethasone groups. Blood samples were collected on days 0, 41, and 55. Peripheral blood mononuclear cells were isolated gravimetrically and stained for markers and cytokines. Monocytes (CD3-CD4+) and T-helpers (CD3+CD4+) were differentiated as target cells. IL-4 was considered a marker for the pro-M2 phenotype since IL-4 can activate M2 macrophage polarization; IFN-g was considered a marker for the pro-M1 phenotype. Time-related changes in the intracellular expression of IL-4 and IFN-g were studied using flow cytometry. Data were analyzed using nonparametric statistical tests (Mann-Whitney-Wilcoxon) in the R environment for statistical computing. Results and conclusions. Monocytes (like macrophages) underwent reprogramming during the development of modeled sJIA disease. In monocytes of doxycycline and dexamethasone treatment groups, pro-M2 effects were observed earlier (day 41) than in the control group. Pro-M1 effects were observed in monocytes of doxycycline and dexamethasone groups on day 55, as compared with the control group. Characteristic time-related changes of intracellular cytokine expression were described for different groups. Importantly, the differently directed phenotype polarization was observed in sJIA and treatment groups for both monocytes and T-helpers.

scholarly journals Autologous Peripheral Blood Mononuclear Cells for Limb Salvage in Diabetic Foot Patients with No-Option Critical Limb Ischemia

2021 ◽  
Vol 10 (10) ◽  
pp. 2213
Author(s):  
Alessia Scatena ◽  
Pasquale Petruzzi ◽  
Filippo Maioli ◽  
Francesca Lucaroni ◽  
Cristina Ambrosone ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Critical Limb Ischemia ◽  
Diabetic Foot ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Limb Ischemia ◽  
Control Group ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Peripheral blood mononuclear cells (PBMNCs) are reported to prevent major amputation and healing in no-option critical limb ischemia (NO-CLI). The aim of this study is to evaluate PBMNC treatment in comparison to standard treatment in NO-CLI patients with diabetic foot ulcers (DFUs). The study included 76 NO-CLI patients admitted to our centers because of CLI with DFUs. All patients were treated with the same standard care (control group), but 38 patients were also treated with autologous PBMNC implants. Major amputations, overall mortality, and number of healed patients were evaluated as the primary endpoint. Only 4 out 38 amputations (10.5%) were observed in the PBMNC group, while 15 out of 38 amputations (39.5%) were recorded in the control group (p = 0.0037). The Kaplan–Meier curves and the log-rank test results showed a significantly lower amputation rate in the PBMNCs group vs. the control group (p = 0.000). At two years follow-up, nearly 80% of the PBMNCs group was still alive vs. only 20% of the control group (p = 0.000). In the PBMNC group, 33 patients healed (86.6%) while only one patient healed in the control group (p = 0.000). PBMNCs showed a positive clinical outcome at two years follow-up in patients with DFUs and NO-CLI, significantly reducing the amputation rate and improving survival and wound healing. According to our study results, intramuscular and peri-lesional injection of autologous PBMNCs could prevent amputations in NO-CLI diabetic patients.

scholarly journals Single-cell characterization of a model of poly I:C-stimulated peripheral blood mononuclear cells in severe asthma

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Ailu Chen ◽  
Maria P. Diaz-Soto ◽  
Miguel F. Sanmamed ◽  
Taylor Adams ◽  
Jonas C. Schupp ◽  
...  
Keyword(s):  
T Cells ◽  
Single Cell ◽  
Severe Asthma ◽  
Mononuclear Cells ◽  
Effector T Cells ◽  
Poly I:C ◽  
Peripheral Blood Mononuclear ◽  
Main Cell ◽  
Blood Mononuclear Cells ◽  
Quantitative Changes

Abstract Background Asthma has been associated with impaired interferon response. Multiple cell types have been implicated in such response impairment and may be responsible for asthma immunopathology. However, existing models to study the immune response in asthma are limited by bulk profiling of cells. Our objective was to Characterize a model of peripheral blood mononuclear cells (PBMCs) of patients with severe asthma (SA) and its response to the TLR3 agonist Poly I:C using two single-cell methods. Methods Two complementary single-cell methods, DropSeq for single-cell RNA sequencing (scRNA-Seq) and mass cytometry (CyTOF), were used to profile PBMCs of SA patients and healthy controls (HC). Poly I:C-stimulated and unstimulated cells were analyzed in this study. Results PBMCs (n = 9414) from five SA (n = 6099) and three HC (n = 3315) were profiled using scRNA-Seq. Six main cell subsets, namely CD4 + T cells, CD8 + T cells, natural killer (NK) cells, B cells, dendritic cells (DCs), and monocytes, were identified. CD4 + T cells were the main cell type in SA and demonstrated a pro-inflammatory profile characterized by increased JAK1 expression. Following Poly I:C stimulation, PBMCs from SA had a robust induction of interferon pathways compared with HC. CyTOF profiling of Poly I:C stimulated and unstimulated PBMCs (n = 160,000) from the same individuals (SA = 5; HC = 3) demonstrated higher CD8 + and CD8 + effector T cells in SA at baseline, followed by a decrease of CD8 + effector T cells after poly I:C stimulation. Conclusions Single-cell profiling of an in vitro model using PBMCs in patients with SA identified activation of pro-inflammatory pathways at baseline and strong response to Poly I:C, as well as quantitative changes in CD8 + effector cells. Thus, transcriptomic and cell quantitative changes are associated with immune cell heterogeneity in this model to evaluate interferon responses in severe asthma.

scholarly journals Release of Cyclooxygenase-2 and Lipoxin A4 from Blood Leukocytes in Aspirin-exacerbated Respiratory Disease

2016 ◽  
Vol 7 (3) ◽  
pp. ar.2016.7.0172
Author(s):  
Ajnacska Rozsasi ◽  
Akos Heinemann ◽  
Tilman Keck
Keyword(s):  
Respiratory Disease ◽  
Mononuclear Cells ◽  
Cyclooxygenase 2 ◽  
Control Group ◽  
Peripheral Blood Mononuclear ◽  
Control Subjects ◽  
Aspirin Exacerbated Respiratory Disease ◽  
Blood Mononuclear Cells ◽  
Cox 2 ◽  
The Difference

Background The release of cyclooxygenase-2 (COX-2) and lipoxin A4 (LXA4) from blood mononuclear cells in patients with aspirin-exacerbated respiratory disease (AERD) is only partially understood. Objective To investigate the presence of COX-2 and LXA4 in peripheral blood mononuclear cells (PBMC) derived from patients with AERD and with nasal polyps (NP) (designated as the AERD-NP group), patients with NP without AERD (the NP group), and healthy controls without sinus disease (the control group). Methods Blood was taken from 14 patients in the AERD-NP group, 6 patients in the NP group, and 8 healthy subjects in the control group. After culturing of human PBMC, the presence of COX-2 protein and LXA4 (ELISA) was detected in the supernatant, and the results were compared among the groups. Results COX-2 and LXA4 were detectable after culturing of PBMC in all patients in the AERD-NP and NP groups and in the control subjects. COX-2 was highest in the patients in the AERD-NP group, but the difference was not significant compared with patients with non-AERD polyp and with the control subjects. LXA4 was also highest in the AERD-NP group, but the difference was also not significant compared with the patients who were non-AERD polyp and the control subjects. Conclusion Neither the release of COX-2 or LXA4 was different between the patients with AERD and with NPs, the patients without AERD and with NPs, and the healthy control group. The release of these proteins in AERD needs further investigation.

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