The immunocytes microglia in the central nervous system (CNS) were reported to play a crucial role in neurodegeneration. As a member of P2 receptors family, purinoceptor P2Y6 has attracted much attention recently. Previous studies showed that purinoceptor P2Y6 mainly contributed to microglia activation and their later phagocytosis in CNS, while in immune system, it participated in the secretion of interleukin (IL)-8 from monocytes and macrocytes. So there raises a question: whether purinoceptor P2Y6 also takes part in neuroinflammation? Thus, this review mainly concerns about the properties and roles of purinoceptor P2Y6, including (1) structure of purinoceptor P2Y6; (2) distribution and properties of purinoceptor P2Y6; (3) relationships between purinoceptor P2Y6 and microglia; (4) relationships between purinoceptor P2Y6 and immunoinflammation. Itos proposed that purinoceptor P2Y6 may play a role in neuroinflammation in CNS, although further research is still required.
Animals ; Humans ; Inflammation ; immunology ; metabolism ; Microglia ; drug effects ; metabolism ; Monocytes ; metabolism ; Phagocytosis ; physiology ; Receptors, Purinergic P2 ; chemistry ; genetics ; metabolism

Cell Hypoxia ; physiology ; Cells, Cultured ; Diabetic Retinopathy ; metabolism ; Female ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Middle Aged ; Monocytes ; metabolism ; Prospective Studies ; Vascular Endothelial Growth Factors ; metabolism

In order to determine the precise mechanism of the interactions between different types of cells, which are common phenomena in tissues and organs, the importance of coculture techniques are becoming increasingly important. In the area of cardiology, artificial arteries have been developed, based on the understanding of physiological communication of the arterial smooth muscle cells (SMC), endothelial cells (EC), and the extracellular matrix (ECM). In the study of atherosclerosis, the modification of low-density lipoprotein (LDL), which result in the recruitment and accumulation of white blood cells, especially, monocytes/macrophages, and foam cell formation, are hypothesized. Although there are well known animal models, an in vitro model of atherogenesis with a precisely known atherogenesis mechanism has not yet been developed. In this paper, an arterial wall reconstruction model using rabbit primary cultivated aortic SMCs and ECs, was shown. In addition, human peripheral monocytes were used and the transmigration of monocytes was observed by scanning electron and laser confocal microscopy. Monocyte differentiation into macrophages was shown by immunohistochemistry and comprehensive gene expression analysis. With the modified form of LDL, the macrophages were observed to accumulate lipids with a foamy appearance and differentiate into the foam cells in the ECM between the ECs and SMCs in the area of our coculture model.
Animal ; Aorta/physiology* ; Aorta/cytology* ; Arteriosclerosis/etiology* ; Cell Differentiation/physiology ; Cell Movement ; Coculture ; Endothelium, Vascular/physiology ; Endothelium, Vascular/cytology ; Extracellular Matrix/metabolism ; Foam Cells/ultrastructure ; Foam Cells/cytology ; Macrophages/physiology* ; Macrophages/cytology ; Male ; Microscopy, Confocal ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Monocytes/ultrastructure ; Monocytes/physiology ; Muscle, Smooth, Vascular/physiology ; Muscle, Smooth, Vascular/cytology ; Myosin/metabolism ; Protein Isoforms/metabolism ; Rabbits

It is generally accepted that tissue factor plays an important role in coagulation and intravascular thrombus formation. Tissue factor is not only found primarily on the surface of certain cells that are located outside the vasculature, but also found in circulating cells. Monocyte express tissue factor induced by endotoxin. Recently, many researches indicate that P-selectin, CD40 ligand and GPIIb/IIIa receptor of platelet can also affect expression of tissue factor by monocyters. In addition, a lot of studies showed that tissue factor exist in the circulation including contained platelet. Tissue factor in the platelet releases under certain condition, and initiates coagulation. In this review the relation between platelet and tissue factor was elaborated.
Blood Platelets ; drug effects ; metabolism ; physiology ; CD40 Ligand ; physiology ; Humans ; Monocytes ; drug effects ; metabolism ; P-Selectin ; physiology ; Peptides ; pharmacology ; Platelet Glycoprotein GPIIb-IIIa Complex ; physiology ; Thromboplastin ; biosynthesis ; drug effects ; physiology

OBJECTIVE: To explore the role of cell-surface nucleolin in lipopolysaccharide (LPS)-stimulated expression and secretion of TNF-alpha and IL-1beta in human THP-1 monocytes. METHODS: Immuno-fluorescence assay and Western blot were used to identify the expression of nucleolin on the surface of THP-1 monocytes. Inactivation of nucleolin was induced by anti-nucleolin monoclonal antibody blockage, and the expression and secretion of TNF-alpha and IL-1beta were observed by using reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immuno-sorbent assay (ELISA)respectively in LPS-mediated human THP-1 monocyte inflammatory model. RESULTS: Immuno-fluorescence showed that nucleolin was localized on the cell surface of THP-1 monocytes as indicated by dotted red fluorescence. Western blot assay indicated that nucleolin existed in the cell membrane fractions. RT-PCR assay showed that the expressions of TNF-alpha and IL-1beta mRNA significantly increased at 2 h and 3 h after the treatment with 1000 microg/L LPS. After 1 h pretreatment with anti-nucleolin antibody, the levels of TNF-alpha and IL-1beta mRNA decreased compared with an anti-nucleolin antibody untreated group and an irrelevant IgG+LPS group (P<0.05). ELISA assay showed that the pretreatment with anti-nucleolin antibody inhibited significantly the secretion of LPS-induced levels of TNF-alpha and IL-1beta after 4, 12 and 24 h treatment with 1000 microg/L LPS. CONCLUSION Nucleolin expresses on the cell surface of THP-1 monocytes and involves in the LPS-mediated expression and secretion of TNF-alpha and IL-1beta.
Cell Line ; Cell Membrane ; metabolism ; Humans ; Interleukin-1beta ; biosynthesis ; metabolism ; Lipopolysaccharides ; pharmacology ; Monocytes ; cytology ; metabolism ; Phosphoproteins ; metabolism ; physiology ; RNA-Binding Proteins ; metabolism ; physiology ; Tumor Necrosis Factor-alpha ; biosynthesis ; metabolism

AIMTo observe the property alteration of K(v) channel in SLE patient's peripheral blood lymphocyte and its significant.

METHODSThe patch-clamp technique was used to record the current of K(V) channel in SLE patient's peripheral lymphocyte.

RESULTSThe current amplitude of K(V) channel in the SLE patient's lymphocytes decreased, it was (258.6 +/- 112.5) pA in healthy people, but in SLE patient it was (139.4 +/- 58.5) pA (P < 0.05). There was no other changes in the property of channel, include activation potential, inactivation property, channel closing kinetics and its pharmacological property.

CONCLUSIONThe decline of SLE patient's cell immunity may be related to the decrease of the amplitude of K(V) channel current.

Case-Control Studies ; Cell Membrane ; physiology ; Flow Cytometry ; Humans ; Lupus Erythematosus, Systemic ; blood ; pathology ; physiopathology ; Lymphocyte Count ; Lymphocytes ; physiology ; Monocytes ; metabolism ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Voltage-Gated ; physiology

OBJECTIVERecent studies have shown that Toll-like receptor 4 (TLR4), a mediator of for innate immune responses, is involved in the initiation and progression of atherosclerosis. TLR4 activation mediates the expression of chemokines and cytokines through activation of NF-kappaB. We investigated the expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), intercellular adhesion molecule-1 (CAM-1), E-selectin induced by TLR4/NF-kappaB in human umbilical vein endothelial cells (HUVECs), and their effects on adhesion of monocyte to HUVECs.

METHODSHUVECs were incubated with purified LPS for 24 h. TLR4, LOX-1, ICAM-1, E-selectin mRNA were measured by RT-PCR; the protein expression of TLR4, LOX-1 and activation of NF-kappaB were detected by Western blot; the adhesive percentage between HUVECs and monocytes was determined by direct counting.

RESULTSLPS (1 mg/L) not only enhanced expression of TLR4, activation of NF-kappaB and induction of LOX-1, ICAM-1, E-selectin expression, but also increased the percentage of monocyte adhesion to endothelium. Pretreatment of HUVECs with anti-LOX1, anti-ICAM-1 or anti E-selectin antibodies partly abolished the increase in monocyte adhesion to endothelium. NF-kappaB inhibitor CAPE suppressed LPS-induced these effects.

CONCLUSIONTLR4/NF-kappaB plays an important role in monocyte-endothelium adhesion partly through upregulation of LOX-1, ICAM-1 and E-selection expression, which may provide a target for the treatment of atherosclerosis.

Cell Adhesion ; Cells, Cultured ; E-Selectin ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Monocytes ; metabolism ; physiology ; NF-kappa B ; metabolism ; Scavenger Receptors, Class E ; metabolism ; Toll-Like Receptor 4 ; metabolism ; Umbilical Veins ; cytology

OBJECTIVETo investigate the role of lysine-specific demethylase 1 (LSD1) in the process of THP-1 monocyte-to-macrophage differentiation.

METHODSQuantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting were performed to analyze the expression of LSD1 and interleukin-6 (IL-6) in THP-1 monocytes and THP-1-derived macrophages. Chromatin immunoprecipitation (ChIP) assay was applied to detect the occupancy of LSD1 and H3K4 methylation at IL-6 promoter during THP-1 monocyte-to-macrophage differentiation. IL-6 mRNA level and H3K4 methylation at IL-6 promoter were analyzed using qRT-PCR and ChIP assay in LSD1-knockdown THP-1 cells treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) for 0, 4, 8, 12, and 24 hours. Fluorescence activated flow cytometry was performed to reveal the percentage of macrophages differentiated from THP-1 monocytes.

RESULTSThe expression of LSD1 reduced during THP-1 monocyte-to-macrophage differentiation (P<0.01). LSD1 occupancy decreased and H3K4 methylation increased at IL-6 promoter during the differentiation. With knockdown of LSD1, H3K4 methylation at IL-6 promoter was found increased after TPA treatment at different times points (all P<0.05, except 24 hours). The percentage of macrophages increased significantly in the THP-1 cells with LSD1 knockdown (P<0.05).

CONCLUSIONSLSD1 is repressed during the monocyte-to-macrophage differentiation of THP-1 cells. Suppression of LSD1-mediated H3K4 demethylation may be required for THP-1 monocyte-to-macrophage differentiation.

Cell Differentiation ; Cells, Cultured ; Dealkylation ; Histone Demethylases ; physiology ; Histones ; metabolism ; Humans ; Interleukin-6 ; genetics ; Macrophages ; cytology ; Monocytes ; cytology ; Promoter Regions, Genetic

OBJECTIVETo observe monocyte (Mo) development in wild type C57BL/6 mice and apoE gene knockout (apoE(-/-)) mice, and to evaluate the immuno-regulatory effect of Huanglian Jiedu Decoction (HJD) on peripheral Mo development in apoE(-/-) mice.

METHODSFour, 8, 12, and 16 weeks old female C57BL/6 mice were set up as control groups of different ages, while 4, 8, 12, and 16 weeks old female apoE(-/-) mice were set up as hyperlipidemia groups of different ages. Four-week old female C57BL/6 mice were recruited as a blank group. Four-week old female apoE(-/-) mice were randomly divided into the control group, the Western medicine group, and the Chinese medicine group by paired comparison, 5 in each group. Equivalent clinical dose was administered to mice according to body weight. Mice in the Western medicine group were administered with Atrovastatin at the daily dose of 10 mg/kg by gastrogavage, while those in the Chinese medicine group were administered with HJD at the daily dose of 5 g/kg by gastrogavage. Body weight was detected each week. After 4 weeks blood lipids levels (such as TG, TC, LDL-C, and HDL-C), and the proportions of Mo and Ly6c(hi) were detected.

RESULTSCompared with 4-week-old homogenic mice, the proportion of Mo decreased in 16-week-old C57BL/6 mice (P < 0.05). Levels of TC and TG, and the proportion of Ly6c(hi) subtype increased, but the proportion of Mo de- creased in 8-week-old apoE(-/-) mice (P <0. 05). Levels of TC, TG, and LDL-C increased in 12-week-old apoE(-/-) mice (P < 0.05). Levels of TC, TG, LDL-C, and HDL-C increased in 16-week-old apoE(-/-) mice (P < 0.05, P < 0.01). Compared with 8-week-old homogenic mice, the proportion of Mo decreased in 16-week-old C57BL/6 mice (P < 0.05); levels of TC and LDL-C increased in 12-week-old apoE(-/-) mice (P < 0.05); levels of TC and HDL-C increased in 16-week-old apoE(-/-) mice (P < 0.05, P < 0.01). Compared with C57BL/6 mice of the same age, TC and TG increased, HDL-C decreased (P < 0.01) in 4-and 8-week-old apoE(-/-) mice (P < 0.01); levels of TC, TG, LDL-C increased, and HDL-C level decreased in 12- and 16-week-old apoE(-/-) mice (P < 0.05, P < 0.01); the proportion of Mo increased in 4-week-old apoE(-/-) mice (P < 0.05); proportions of Mo and Ly6c(hi) increased in 8-week-old apoE(-/-) mice (P < 0.05). Compared with the blank control group, levels of TC, TG, and LDL-C, proportions of Mo and Ly6c(hi) increased (P < 0.01, P < 0.05), but HDL-C level decreased (P <0. 01) in the control group after intervention. Compared with the control group, body weight gained less in the Western medicine group and the Chinese medicine group (P < 0.05); the proportion of Ly6c(hi) subtype decreased in the Chinese medicine group (P < 0.05).

CONCLUSIONSIn development process blood lipids levels in apoE(-/-) mice are not only associated with age. Blood lipids levels induced growth changes in natural immune system are also correlated with age. In early stage of lipids development HJD intervention could correct this special immune disorder in apoE(-/-) mice.

Animals ; Apolipoproteins E ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Knockout Techniques ; Hyperlipidemias ; Lipids ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Monocytes ; physiology

Objective: To investigate the relationship between human cytomegalovirus (HCMV) infection and peripheral blood CD14 CD16 monocytes in the pathogenesis of coronary heart disease (CHD), and to elucidate the mechanism of pathogenesis in CHD by analyzing the correlation between infection, inflammation, and CHD, to provide a basis for the prevention, evaluation, and treatment of the disease. Methods: In total, 192 patients with CHD were divided into three groups: latent CHD, angina pectoris, and myocardial infarction. HCMV-IgM and -IgG antibodies were assessed using ELISA; CD14 CD16 monocytes were counted using a five-type automated hematology analyzer; mononuclear cells were assessed using fluorescence-activated cell sorting; and an automatic biochemical analyzer was used to measure the levels of triglyceride, cholesterol, high- and low-density lipoprotein cholesterols, lipoprotein, hs-CRp and Hcy. Results: The positive rates of HCMV-IgM and -IgG were significantly higher in the CHD groups than in the control group. HCMV infection affects lipid metabolism to promote immune and inflammatory responses. Conclusion HCMV infection has a specific correlation with the occurrence and development of CHD. The expression of CD14 CD16 mononuclear cells in the CHD group was increased accordingly and correlated with acute HCMV infection. Thus, HCMV antibody as well as peripheral blood CD14 CD16 mononuclear cells can be used to monitor the occurrence and development of CHD.
Angina Pectoris ; epidemiology ; virology ; China ; epidemiology ; Coronary Disease ; epidemiology ; virology ; Cytomegalovirus ; physiology ; Cytomegalovirus Infections ; complications ; Humans ; Incidence ; Inflammation ; epidemiology ; etiology ; Leukocyte Count ; Monocytes ; metabolism ; Myocardial Infarction ; epidemiology ; virology