Cells, Cultured ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Lung ; cytology ; metabolism ; Macrophages, Alveolar ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Silicon Dioxide ; toxicity ; Silicosis ; metabolism

The role of very low density lipoprotein receptor (LVLDR) in the process of foam cell formation was investigated. After the primary cultured mouse peritoneal macrophages were incubated with VLDL, beta-VLDL or low density lipoprotein (LDL), respectively for 24 h and 48 h, foam cells formation was identified by oil red O staining and cellular contents of triglyceride (TG) and total cholesterol (TC) were determined. The mRNA levels of LDLR, LDLR related protein (ILRP) and VLDLR were detected by semi-quantitative RT-PCR. The results demonstrated that VLDL, beta-VLDL and LDL could increase the contents of TG and TC in macrophages. Cells treated with VLDL or beta-VLDL showed markedly increased expression of VLDLR and decreased expression of LDLR, whereas LRP was up-regulated slightly. For identifying the effect of VLDL receptor on cellular lipid accumulation, ldl-A7-VR cells, which expresses VLDLR and trace amount of LRP without functional LDLR, was used to incubate with lipoproteins for further examination. The results elucidated that the uptake of triglyceride-rich lipoprotein mediated by VLDLR plays an important role in accumulation of lipid and the formation of foam cells.
Animals ; Arteriosclerosis ; metabolism ; pathology ; Cells, Cultured ; Cholesterol, LDL ; metabolism ; pharmacology ; Female ; Foam Cells ; cytology ; metabolism ; Lipoproteins, VLDL ; pharmacology ; Macrophages, Peritoneal ; cytology ; metabolism ; Mice ; Receptors, LDL ; metabolism ; Triglycerides ; metabolism

Animals ; Arthritis, Experimental ; metabolism ; pathology ; Breast Neoplasms ; pathology ; Cadherins ; metabolism ; physiology ; Cell Movement ; Female ; Fibroblasts ; cytology ; pathology ; Humans ; Macrophages ; cytology ; pathology ; Neoplasm Invasiveness ; Synovial Membrane ; cytology ; metabolism ; pathology

Bronchoalveolar Lavage Fluid ; Humans ; Interleukin-1beta ; metabolism ; Macrophages, Alveolar ; cytology ; drug effects ; metabolism ; Silicon Dioxide ; adverse effects ; Silicosis ; metabolism

OBJECTIVETo observe the inhibitory effect of oligodeoxynucleotides (ODN) on nuclear translocation and nuclear binding activity of nuclear factor (NF)-kappaB in THP-1 cells.

METHODSOligodeoxynucleotides were transfected via liposome into THP-1 cells followed by stimulation of the cells with lipopolysaccharide (LPS). Immunocytochemistry, electrophoretic mobility shift assay (EMSA) and reverse transcription (RT)-PCR were performed to detect the nuclear translocation and nuclear binding activity of NF-kappaB.

RESULTSImmunocytochemical results showed that after LPS stimulation of the ODN-transfected cells, NF-kappaB expression was still localized in cytoplasma. EMSA demonstrated inhibited nuclear binding activity of NF-kappaB in the ODN-transfected cells, and ODN inhibited the mRNA expression of tumor necrosis factor (TNF)-alpha, a NF-kappaB-associated inflammatory factor, as shown by RT-PCR.

CONCLUSIONODN can inhibit the nuclear translocation and binding activity of NF-kappaB in THP-1 cells, whereby the transcription and expression of the related inflammation factor genes is suppressed, which shed light on a new solution for clinical treatment of acute pancreatitis.

Cell Line ; Humans ; Lipopolysaccharides ; Macrophages ; cytology ; drug effects ; metabolism ; NF-kappa B ; metabolism ; Oligodeoxyribonucleotides ; pharmacology ; Transcription Factors ; metabolism ; Transfection

OBJECTIVETo investigate the effect of macrophages on embryo implantation by observing the distribution of macrophages in mouse uterine tissues during the peri-implantation period.

METHODSUterine tissues were collected from pregnant (n=30) and pseudopregnant mice (n=30) during the peri-implantation period. The distributions of macrophages, iNOS and leukemia inhibitory factor (LIF) were determined by immunohistochemistry and the correlations of macrophages with iNOS and LIF were analyzed.

RESULTSMacrophages were located mainly in the endometrium before D4.5 in the pregnant rats with D0.5 defined as the morning when a vaginal plug was observed. After D4.5, the macrophages was significantly reduced in number (P<0.05) in the endometrium and gradually migrated to the perimetrium. In the psudopregnant mice, macrophages were located mainly in the endometrium. Before D4.5, iNOS-positive cells were detected mainly in the endometrium and the myometrium in the pregnant rats and became significantly reduced on D4.5 (P<0.05); in the pseudopregnant mice, the positive cells were mostly detected in the endometrium. Significant differences were found in the distribution of the macrophages and LIF between the implantation and non-implantation sites (P=0.013). LIF was mostly located in the endometrium in the pregnant mice but scarcely detected in the pseudopregnant mice.

CONCLUSIONMacrophages are located mainly in the endometrium and the implantation site where iNOS and LIF are expressed, suggesting the important role of macrophages in the determination of implantation.

Animals ; Blood Cell Count ; Embryo Implantation ; Endometrium ; cytology ; Female ; Immunohistochemistry ; Leukemia Inhibitory Factor ; metabolism ; Macrophages ; cytology ; Mice ; Nitric Oxide Synthase Type II ; metabolism ; Pregnancy ; Uterus ; cytology

OBJECTIVETo establish the co-culture model of human macrophage cell line (U937) with human vein umbilical cell line (ECV304), and to explore the feasibility of using concanavalin A (ConA) as U937 cell stimulator in regulating angiogenesis.

METHODSECV304 cells were cultured in vitro, and to which were respectively added U937 cells (1 x 10(5)), 25 microg/mL ConA, and U937 cell (1 x 10(5)) + ConA (25 microg/mL) after cell fusion rate reaching 60%, and then co-cultured for 48 hours. ECV 304 cells in conventional culture were used as controls. 3H-TdR incorporation test was employed to determine the DNA synthesis of vascular endothelial cells. Flow cytometry was used to determine the changes in the cell cycle, and RT-PCR was adopted to determine the expression of homeobox (HOXB2) mRNA.

RESULTSAfter conA stimulation to ECV 304 co-cultured with U937 cells, the percentage of cells in S phase (48.860 +/- 2.290), the DNA synthesis [(5694 +/- 917) min(-1)], and the expression of HOXB2 mRNA (0.947 +/- 0.003) were obviously higher than those in control group [41.590 +/- 2.590 vs (2498 +/- 1109) min(-1) vs 0.646 +/- 0.004, P > 0.01]. There was no obvious difference in apoptosis among above stimulation methods (P >0.05).

CONCLUSIONU937 cells activated by ConA can promote the proliferation of ECV304 cells and further regulate angiogenesis. HOXB2 gene is closely related to the endothelial proliferation.

Apoptosis ; Cell Division ; Cell Line ; Coculture Techniques ; Endothelial Cells ; cytology ; metabolism ; Endothelium, Vascular ; cytology ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Macrophages ; cytology ; metabolism ; RNA, Messenger ; metabolism ; Transcription Factors ; genetics ; metabolism ; Umbilical Veins ; cytology

To explore the anti-atherosclerotic mechanism of estrogen and especially observe the effect of estradiol on the content of cholesterol in J774a.1 mouse mononuclear/macrophage-derived foam cells which were incubated with oxidized low-density lipoproteins (ox-LDL). J774a.1 mouse mononuclear/macrophages were incubated with ox-LDL or with both ox-LDL and estradiol (1, 0.1 or 0.01 micromol x L(-1)). Oil red O staining was used to observe the formation of foam cells, and cholesterol oxidase fluorometric was used to determine the content of cellular cholesterol content. Western blotting and RTFQ-PCR were used to observe the expressions of scavenger receptor class B type I (SR-B I ) in J774a.1 foam cells. Compared with the control cells, J774a.1 mouse mononuclear/macrophage-derived foam cells showed significantly increased contents of total cholesterol and cholesterol ester (P < 0.001) and decreased SR-B I mRNA expression (P < 0.01). Estradiol treatment significantly lowered the contents of total cholesterol and cholesterol ester (P < 0.05), and increased SR-B I protein and mRNA expression (P < 0.01) in the foam cells in a dose-dependent manner. Estradiol can inhibit the formation of mononuclear/macrophage-derived foam cells by decreasing the contents of total cholesterol and cholesterol ester and up-regulating the expression of SR-B I in the foam cells.
Animals ; Cell Line ; Cholesterol ; metabolism ; Cholesterol Esters ; metabolism ; Estradiol ; pharmacology ; Foam Cells ; cytology ; metabolism ; Lipoproteins, LDL ; metabolism ; Macrophages ; drug effects ; metabolism ; Mice ; Scavenger Receptors, Class B ; metabolism

OBJECTIVETo compare the serum miR-663 levels in renal transplant patients with and without acute rejection (AR) and explore the role of miR-663 acute renal graft rejection.

METHODSReal time-PCR was used to determine serum miR-663 levels in renal transplant recipients with and without AR. MTT assay and Annexin V-FITC assay were employed to examine the viability and apoptosis of human renal glomerular endothelial cells (HRGEC) treated with a miR-663 mimic or a miR-663 inhibitor, and ELISA was performed to detect the expression of inflammation-related cytokines including IL-6, IFN-γ, CCL-2 and TNF-α in the cells. Transwell assay was used to examine the effect of miR-663 mimic and miR-663 inhibitor on the chemotactic capability of macrophages.

RESULTSSerum miR-663 level was significantly higher in renal transplant recipients with AR than in those without AR. The miR-663 mimic significantly inhibited the viability of HRGECs and increase the cell apoptosis rate, while miR-663 inhibitor suppressed the cell apoptosis. The miR-663 mimic increased the expression levels of inflammation-related cytokines and enhanced the chemotactic capability of macrophages.

CONCLUSIONmiR-663 might play important roles in acute renal graft rejection and may become a therapeutic target for treating AR.

Apoptosis ; Cells, Cultured ; Cytokines ; metabolism ; Endothelial Cells ; cytology ; Graft Rejection ; blood ; Humans ; Kidney Glomerulus ; cytology ; Kidney Transplantation ; Macrophages ; cytology ; drug effects ; MicroRNAs ; blood

Atherosclerosis is a chronic, inflammatory disorder characterized by the deposition of excess lipids in the arterial intima. The formation of macrophage-derived foam cells in a plaque is a hallmark of the development of atherosclerosis. Lipid homeostasis, especially cholesterol homeostasis, plays a crucial role during the formation of foam cells. Recently, lipid droplet-associated proteins, including PAT and CIDE family proteins, have been shown to control the development of atherosclerosis by regulating the formation, growth, stabilization and functions of lipid droplets in macrophage-derived foam cells. This review focuses on the potential mechanisms of formation of macrophage-derived foam cells in atherosclerosis with particular emphasis on the role of lipid homeostasis and lipid droplet-associated proteins. Understanding the process of foam cell formation will aid in the future discovery of novel therapeutic interventions for atherosclerosis.
Acyltransferases ; metabolism ; Apoptosis Regulatory Proteins ; metabolism ; Atherosclerosis ; metabolism ; pathology ; Cholesterol ; metabolism ; Foam Cells ; cytology ; metabolism ; Humans ; Lipid Metabolism ; physiology ; Macrophages ; cytology ; immunology ; Membrane Proteins ; metabolism ; Perilipin-2 ; Peroxisome Proliferator-Activated Receptors ; metabolism ; Sterol Regulatory Element Binding Proteins ; metabolism