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Objective Little research has been done on how Cx37 changes the current density of mononuclear macrophage in atherosclerosis.The purpose of this study was to detect the effects of Cx37 on the current density of mononuclear macrophage in atherosclerosis.Methods A total of 30 Wistar mice were randomly divided into Cx37+ group and Cx37-group equally.The atherosclerosis model was constructed by a high-fat diet.According to different parts of sample collection, these two groups were subdivided into Cx37+ plaque group, Cx37-plaque group, Cx37+blood group and Cx37-blood group.RT-PCR was applied to detect the expression of Cx37 in different body parts.The mononuclear macrophages were cultured after being separated from blood and plague in both groups.The current density of mononuclear macrophage was detected by the whole cell recording.Results The relative expression of Cx37 in Cx37 + plaque group was higher than that in plaque group ([1.10±0.02] vs [0.60±0.03]).Energy Spectrum CT was used to detect the carotid artery plaque in both Cx37 + and Cx37-groups, which verified the successful model construction.At 80,120 and 160ms, the current density in Cx37 + plaque group([0.61± 0.06], [0.67±0.07], [0.91±0.03]A/cm2) was significantly higher than those in Cx37 + blood group([0.49±0.02], [0.61±0.03], [0.67±0.02]A/cm2) , Cx37-plaque group([0.48±0.02], [0.60±0.02], [0.64±0.02]A/cm2) and Cx37-blood group([0.49±0.02], [0.59±0.02], [0.64±0.02]A/cm2).The same goes for those at 200, 240, 320ms(P<0.05).Conclusion Cx37 has more significant impact on the current density in the plaque of mononuclear macrophage than in the peripheral blood in promoting macrophages activation and atherosclerosis progress.
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Objective To understand the differences between infiltration of mononuclear-macro-phages and neutrophils during Leptospira interrogans (L.interrogans) infection and the underlying mecha-nisms. Methods Histological changes in mouse lung, liver and kidney tissues were detected using hema-toxylin and eosin (HE) staining following infection of C3H/HeJ mice with L.interrogans serovar Lai strain 56601. Infiltration of peripheral blood-derived CD11b+mononuclear-macrophages and Ly6G+neutrophils in lung,liver and kidney tissues collected form L.interrogans-infected C3H/HeJ mice was detected with immu-nohistochemistry. Levels of mononuclear-macrophage chemokines and neutrophil chemokines in serum sam-ples of L.interrogans-infected mice were detected with chemokine detection microarray. Results Lung,liv-er and kidney tissue samples collected from L. interrogans-infected C3H/HeJ mice presented typical his-topathological changes of leptospirosis, such as inflammatory cell infiltration in these tissues, pulmonary hemorrhage,extensive hepatocyte necrosis and serious nephrohemia. Results of immunohistochemical stai-ning showed that a large number of peripheral blood-derived CD11b+mononuclear-macrophages were presen-ted in lung,liver and kidney tissues of L.interrogans-infected mice, but few neutrophils could be found in these tissues. The mouse chemokine detection microarray confirmed that the levels of mononuclear-macro-phage chemokines (I-309,MCP-1,MCP-5,MIP-1α and RANTES) in serum samples of L.interrogans-in-fected C3H/HeJ mice were significantly increased during infection (P<0.05), but the neutrophil chemokines(KC,LIX and MIP-2) analyzed in this study were not notably increased (P>0.05). Conclu-sion Mononuclear-macrophages rather than neutrophils are the major infiltrating phagocytes during L.inter-rogans infection and play a crucial role in the elimination of Leptospira invasion.
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Objective To understand the differences in killing ability and mechanism of human mononuclear macrophages and neutrophils against Leptospira interrogans.Methods Human THP-1 and HL-60 cell lines were respectively pretreated with PMA (phorbol 12-myristate 13-acetate) and ATRA (all-trans retinoic acid) to induce their differentiation into macrophages and neutrophils.Confocal microscopy was used to detect the changes in total ROS (reactive oxygen species) and NO (nitric oxide) levels as well as free Ca2+ concentration ([Ca2+]i) in THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils after infection with Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai.Fluorospectrophotometry was applied to analyze the differences in killing ability between THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils against intracellular leptospires before and after treatment with total ROS and NO inhibitors and intracellular free Ca2+ chelator.Results The total ROS and NO levels and [Ca2+]i in THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils were significantly increased after infection with the spirochete (P<0.05).Moreover,the total ROS and NO levels and [Ca2+]i in the former were significantly higher than those in the latter (P<0.05).The THP-1 monocyte-derived macrophages had stronger killing ability against intracellular leptospires than the HL-60 cell-derived neutrophils (P<0.05).Inhibiting total intracellular ROS,NO or free Ca2+ could result in decreased killing ability of THP-1 monocyte-derived macrophages against intracellular leptospires,but not affect the killing ability of HL-60 cell-derived neutrophils.Conclusion Mononuclear macrophages rather than neutrophils act as the main phagocytes eliminating Leptospira interrogans.High levels of total intracellular ROS,NO and free Ca2+ are closely associated with the ability of mononuclear macrophages to kill Leptospira interrogans.
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Objective To investigate expression differences of neutrophil and mononuclear phagocyte related gene mRNAs among acute myocar-dial infarction (AMI), stable angina (SA) and control groups, and then discuss their expression characteristics in the stable angina pectoris (SAP) and AMI stages of coronary artery disease (CAD). Methods Whole Human Genome Oligo Microarrays were applied to assess the differential expression characteristics of neutrophil and mononuclear phagocyte related mRNAs in patients with AMI (n=20), SA (n=20) and controls (n=20). Results (1) Almost all colony-stimulating factors (CSF) and their receptors related mRNAs was up-regulated in AMI and SA groups compared with the control group, and the expression of granulocyte-macrophage colony stimulating factor receptor (GM-CSFR) and granulocyte colony stimulating factor receptor (G-CSFR) mRNAs in the AMI group was significantly up-regulated com-pared with the other two groups (P<0.01). (2) The expression of mRNAs related to monocyte chemoattractant protein-1 (MCP-1), CCR2 (MCP-1 receptor) and CXCR2 (IL-8 receptor) was significantly up-regulated (P<0.01) in AMI group compared with SA and control groups. IL-8 mRNA expression in the AMI group was clearly higher than the controls (P<0.05). (3) All mRNAs expression related to opsonic re-ceptors (IgG FcR and C3bR/C4bR) was significantly up-regulated in AMI group compared with SA and control group (P<0.01), and the SA group showed an upward trend compared with controls. (4) Most pattern recognition receptor (PRR)-related mRNAs expression was up-regulated in AMI group compared with SA and control groups. Most toll-like receptor (TLR) mRNAs expression was significantly up-regulated (P<0.01) than the SA and control groups;macrophage scavenger receptor (MSR) mRNA was significantly up-regulated in AMI group compared with the control group (P<0.01), and the SA group showed an upward trend compared with the controls. Conclusions The expression of most neutrophil and mononuclear-macrophage function related genes mRNAs was significantly up-regulated by stages during the progression of CAD, suggesting that the adhesive, chemotactic and phagocytic functions of neutrophil and mononuclear-ma-crophage were strengthened in the occurrence and development of coronary atherosclerosis and AMI. This also showed a stepped up-ward trend as the disease progressed.
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Objective To determine the modality of Leptospira interrogans invading human and murine mononuclear-macrophages and diversity of leptospiral phagocytotic vesicle formation. Methods Transmission electron microscopy was applied to observe the invasion of L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai into murine mononuclear-macrophage-like cell line J774A. 1 and PMA-activated human monocyte line THP-1 and the formation of leptospiral phagocytotic vesicles. By using immunofluorescence plus either laser confocal microscopy or fluorescence spectrophotometry, the changes of intracellular leptospiral numbers in J774A. 1 and PMA-activated THP-1 cells before and after block with endocytosis inhibitors monodansylcadaverin (MDC), phenylarsine oxide (PAO) and clathrin antibody were investigated. Results The leptospires in J774A. 1 cells were located in phagocytotic vesicles while the leptospires in THP-1 cells had no package with phagocytotic vesicle membrane. Both MDC and PAO presented the effect inhibiting endocytosis of L. interrogans into J774A. 1 and THP-1 cells in dose-dependent manner. The numbers of leptospires in J774A. 1 and THP-1 cells that pre-blocked with 10 μmol/L or above MDC and 1 μmol/L or above PAO were significantly less than that in the two cells untreated with MDC and PAO (P＜0.05＝. After J774A. 1 and THP-1 cells were blocked with clathrin antibody, the numbers of intracellular leptospires were also remarkbly decreased ( P<0.05 ).Conclusion Leptospira interrogans can invade into both human and murine mononuclear-macrophages through the way of clathrin-dependent endocytosis. There is an opposite diversity of leptospiral phagocytotic vesicle formations in human and murine mononuclear-macrophages, which may result in the difference of pathogenesis in human and mice after infected with L. interrogans.
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Atherosclerosis(AS)is a chronic inflammatory disease of arteries,induced by the injury of endothelial cells,accumulation of mononuclear macrophage and lipid,proliferation of smooth muscle cells and the adhesion of platelets.As a frequent disease and a common pathology in cardiovascular system.In fact,the inciting event of AS is likely an inflammatory insult that occurs decades before the disease becomes clinically apparent.More and more cases were also reported in the childhood recently.Therefore it is important both for clinic and research to investigate the cause and pathogenesis of AS.J Appl Clin Pediatr,2009,24(1):58-60