This study was aimed to explore the expression of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 in bcr/abl fusion gene positive CML cells, and to study the effects of P210(bcr/abl) fusion protein tyrosine kinase on expression of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 mRNAs in chronic myeloid leukemia cells. The expression levels of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 mRNA were detected by semi-quantitative RT-PCR in bcr/abl negative cells, bcr/abl positive cells, and P210(bcr/abl)-Rb-C-Box positive cells. The results showed that MIP-1alpha and CCR-1 mRNAs were expressed in bcr/abl negative cells, but not in positive cells. Both MCP-1 and CCR-2 mRNA cannot be detected in both bcr/abl positive and negative cells. After inhibiting P210(bcr/abl) tyrosine kinase activity by Rb-C-Box, expressions of MIP-1alpha and CCR-1 mRNAs were restored to normal (similar to P210(bcr/abl) negative cells). It is concluded that P210(bcr/abl) fusion protein inhibits the expression of MIP-1alpha and CCR-1 in chronic myeloid leukemia cells, but does not inhibit MCP-1 and CCR-2 mRNA expressions in these leukemia cells.
Chemokine CCL2 ; biosynthesis ; genetics ; Chemokine CCL3 ; Chemokine CCL4 ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Macrophage Inflammatory Proteins ; biosynthesis ; genetics ; Receptors, CCR1 ; Receptors, CCR2 ; Receptors, Chemokine ; biosynthesis ; genetics ; Tumor Cells, Cultured

OBJECTIVE: To explore the microvessel counts and the expressions of interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein-1 ( MIP-1) alpha mRNA in the pathological scar tissues. METHODS: Immunohistochemical method of avidin-biotin complex was used for microvessel counts on the routinely formalin-fixed and paraffin-embedded sections of specimens of hypertrophic scars, keloids, normal skin, and surgical scar, and in situ hybridization for the expressions of IL-8, MCP-1, MIP-1alpha mRNA. RESULTS: The microvessel counts as well as the positive rates and the scorings of IL-8, MCP-1, and MIP-1alpha mRNA were significantly higher in pathological scars than those in the normal skin and surgical scar (all P < 0.05). The microvessel counts were significantly higher in the positive cases of IL-8, MCP-1 and MIP-1alpha mRNA than those in the negative ones (P < 0.05). The close positive correlations were found among the microvessel counts and the expressive scorings of 3 factors (P < 0.05). The close positive correlations were also found among the expressive scorings of IL-8, MCP-1, and MIP-1alpha mRNA in pathological scars. Microvessel counts were significantly higher in hypertrophic scars with the course less than 1 year than those with the course more than 1 year. CONCLUSION IL-8, MCP-1 and MIP-1alpha play important roles in promoting the neovascularization of pathological scars.
Adolescent ; Adult ; Burns ; complications ; Capillaries ; metabolism ; Chemokine CCL2 ; biosynthesis ; genetics ; Chemokine CCL3 ; Chemokine CCL4 ; Cicatrix ; etiology ; metabolism ; Female ; Humans ; Interleukin-8 ; biosynthesis ; genetics ; Macrophage Inflammatory Proteins ; biosynthesis ; genetics ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Skin ; blood supply

OBJECTIVETo explore the role of monocyte chemoattractant protein-1 (MCP-1) in lupus nephritis (LN).

METHODSSera MCP-1 levels were measured by enzyme linked immunosorbent assay in 112 patients with systemic lupus erythematosus (SLE), 30 patients with rheumatoid arthritis, 11 non-SLE patients with renal impairment, and 40 healthy volunteers. MCP-1 mRNA expression in peripheral blood mononuclear cells (PBMCs) was also investigated with reverse trancription-polymerase chain reaction semi-quantitative method.

RESULTSThe expression of MCP-1 was significantly higher in active LN groups than in all other groups (P < 0.001), and there was a close correlation between MCP-1 expression and the overall SLE disease activity index score (r=0.6245, P < 0.001) and the SLE disease activity index renal score (r=0.6808, P < 0.001). Low expression of MCP-1 was observed in diseased controls and healthy controls. The sera levels of MCP-1 were significantly higher in patients with active diseases than in patients with inactive SLE and controls, but no significant difference were found between the active LN groups and non-renal involvement group (P >0.05).

CONCLUSIONThe expression of PBMCs MCP-1 mRNA is upregulated in active SLE. Meanwhile, its expression levels are correlated with the activity of LN.

Adolescent ; Adult ; Aged ; Arthritis, Rheumatoid ; metabolism ; Chemokine CCL2 ; biosynthesis ; genetics ; Female ; Humans ; Lupus Erythematosus, Systemic ; metabolism ; Lupus Nephritis ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo evaluate the possibility and efficiency of nanoparticle as a new vector in specific gene transference.

METHODSNanoparticle-DNA complex was prepared with Poly-dl-lactic-co-glycolic acid (PLGA) bearing anti-sense monocyte chemotactic protein-1 (A-MCP-1), a specific expression gene, and the package efficiency, release progress in vitro, and the size of the complex were determined. The possibility of the new vector was evaluated with genomic DNA PCR by transferring gene into cultured smooth muscle cells (SMC), cationic lipids as a control. For study in vivo, jugular vein-to-artery bypass grafting procedures were performed on 20 New Zealand white rabbits, of which 6 grafts were transferred with nanoparticle-A-MCP-1 (200 microg), 6 with A-MCP-1 (200 microg) by cationic liposome, 4 with LNCX plasmid, and 4 as control. Fourteen days after the grafts were harvested, the expression of A-MCP-1 and its effect on MCP-1 in vein grafts were detected by dot blot, and the morphologic evaluation of grafts was performed.

RESULTSThe package efficiency of the nanoparticle-DNA complex was 0.9%, release progress in vitro lasted 2 weeks, and the size ranged from 150 to 300 nm. SMC genomic DNA PCR showed that A-MCP-1 gene could be successfully transfected into cells by nanoparticle. The study in vivo indicated that A-MCP-1 mRNA was expressed in both local gene delivery groups, nanoparticle and liposome, meanwhile, MCP-1 expression in vein grafts was significantly inhibited and neointimal hyperplasia was notably reduced.

CONCLUSIONNanoparticle can act as a vector to transfect specific gene.

Animals ; Chemokine CCL2 ; biosynthesis ; genetics ; DNA, Antisense ; biosynthesis ; genetics ; Drug Carriers ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Lactic Acid ; Nanotechnology ; Particle Size ; Polyglycolic Acid ; Polymers ; Rabbits ; Transfection

Cells, Cultured ; Chemokine CCL2 ; biosynthesis ; genetics ; Endothelial Cells ; metabolism ; Humans ; Mitogen-Activated Protein Kinases ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; Umbilical Veins ; cytology ; p38 Mitogen-Activated Protein Kinases

Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-kappaB and the expression of chemokine MOB-1, MCP-lgenes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTC in vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-kappaB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10(-3) mol/L, 10(-4) mol/L carbachol) could induce a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10(-3) mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-kappaB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10(-3) mol/L carbachol) in vitro for 30 min. Either M3 cholinergic receptor antagonist (10(-5) mol/L atropine) or NF-kappaB inhibitor (10(-2) mol/L PDTC) could obviously inhibit the activation of NF-kappaB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-lgenes in pancreatic acinar cells in vitro through the activation of NF-kappaB.
Adaptor Proteins, Signal Transducing ; Carbachol ; pharmacology ; Carrier Proteins ; biosynthesis ; genetics ; Chemokine CCL2 ; biosynthesis ; genetics ; Chemokines ; biosynthesis ; genetics ; Humans ; NF-kappa B ; biosynthesis ; genetics ; Pancreas, Exocrine ; metabolism ; Pancreatitis ; etiology ; RNA, Messenger ; biosynthesis ; genetics ; Receptor, Muscarinic M3 ; agonists ; physiology ; Signal Transduction

OBJECTIVETo investigate the expression of monocyte chemotactic protein 1 (MCP-1) in coronary atherosclerosis plaque of sudden coronary death (SCD) patients and the relationship between MCP-1 expression and SCD.

METHODSAutopsy heart samples (n = 90) collected during 2001 - 2003 were divided to SCD group (n = 36) and 2 control groups: control group I, non-SCD CHD (n = 28), control group II, non-cardiac death (n = 26). The immuno-histochemistry SABC techniques (R, positive MCP-1 cell area/totl area) and computerized images analysis (A) were performed to detect the expression of MCP-1 in different groups.

RESULTSR and A in plaques are significant higher in SCD group than control group I and II (0.1264 +/- 0.013 vs 0.0269 +/- 0.0110 and 0.0267 +/- 0.0100, P = 0.04), (0.4534 +/- 0.083 vs 0.2303 +/- 0.040 and 0.2158 +/- 0.0400, P = 0.00), and similar between control group I and control group II.

CONCLUSIONSMCP-1 expression is increased in coronary atherosclerosis plaque of SCD patients.

Adolescent ; Adult ; Aged ; Cadaver ; Chemokine CCL2 ; biosynthesis ; Coronary Artery Disease ; metabolism ; pathology ; Death, Sudden, Cardiac ; pathology ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged

Cytokine and chemokine receptors play a key role in inflammation caused by rheumatoid arthritis (RA). Two isoforms of human CC chemokine receptor R2 (CCR2), the receptor of monocyte chemoattractant protein 1 (MCP-1), have been identified but their relative expression in fibroblast-like synoviocytes (FLS) and their contribution to inflammatory responses mediated by MCP-1 or inflammatory cytokines in patients with RA remain uncertain. We examined the pattern of expression of two CCR2 isoforms upon stimulation by proinflammatory cytokines and CD40 ligation. FLS were prepared from the synovial tissues of RA patients and cultured in the presence of MCP-1, soluble CD40 ligand (sCD40L), TGF-beta, IL-1beta, IL-18, IL-15, and LPS. CCR2A and CCR2B expression was examined by immunohistochemistry, RT-PCR and western blot analysis. IL-15, TNF-alpha and MCP-1 production was determined by ELISA. Immunohistochemistry showed that CCR2A is highly expressed in RA synovium compared with OA synovium. Transcripts of both CCR2A and CCR2B were detected in FLS. Exogenous MCP-1, CD40L, TGF-beta, and IL-15 significantly increased the expression of CCR2A but not CCR2B. Exposure of FLS to sCD40L caused strong upregulation of CCR2A but not of CCR2B protein expression. MCP-1 increased the proliferation of FLS and the production of IL-15, TNF-alpha, and IL-18. Because CCR2A is the main target of regulation by cytokines and CD40 ligation, the relatively higher expression of CCR2A on the cell surface suggests that this isoform of MCP-1 receptor functions as the principal mediator of inflammatory signals in RA FLS.
Arthritis, Rheumatoid/*metabolism ; CD40 Ligand/*pharmacology ; Cells, Cultured ; Chemokine CCL2/*pharmacology ; Chemokines/biosynthesis ; Fibroblasts/*metabolism ; Humans ; Protein Isoforms ; Receptors, CCR2/*biosynthesis ; Synovial Membrane/*pathology ; Transforming Growth Factor beta/*pharmacology

Aim of this article was to investigate relationship between inflammatory pathogenesis of diabetic nephropathy and the TCM pathogenetic theory of Shen-Collateral impaired by Toxin, and to illustrate the method for removing toxin, activating collateral and protecting Shen can be an effective treatment for inhibiting the inflammatory pathogenesis of diabetic nephropathy.
Chemokine CCL2 ; biosynthesis ; genetics ; Diabetic Nephropathies ; drug therapy ; etiology ; metabolism ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Inflammation ; complications ; metabolism ; Male ; Medicine, Chinese Traditional ; NF-kappa B ; biosynthesis ; genetics ; Phytotherapy

AIMTo study the intervention of cetirizine on monocyte chemoattractant protein-1 (MCP-1) in different cutaneous inflammation models.

METHODSHistamine and IFN-gamma stimulated dermal fibroblast cells and HaCaT cells to mimic cutaneous inflammation. Expression of MCP-1 was assessed by means of RT-PCR and ELISA.

RESULTSCompared with the control group of dermal fibroblast (DF) cells and HaCaT cells, MCP-1 mRNA was significantly upregulated by histamine (10 micromol x L(-1)) and IFN-gamma (20 ng x mL(-1)). The protein secretions of MCP-1 were increased 3.5 fold and 8.4 fold in DF cells, respectively. The similar tendency was observed in HaCaT cells. The enhancing effects of histamine and IFN-gamma on MCP-1 protein production were significantly inhibited by cetirizine (1 and 10 micromol x L(-1)) in DF and HaCaT cells.

CONCLUSIONCetirizine may exert the anti-inflammatory effect of skin via inhibiting MCP-1 expression.

Cell Line ; Cells, Cultured ; Cetirizine ; pharmacology ; Chemokine CCL2 ; biosynthesis ; genetics ; Dermatitis ; metabolism ; Dermis ; cytology ; Fibroblasts ; cytology ; metabolism ; Histamine ; pharmacology ; Histamine H1 Antagonists, Non-Sedating ; pharmacology ; Humans ; Interferon-gamma ; pharmacology ; Keratinocytes ; cytology ; metabolism ; RNA, Messenger ; biosynthesis ; genetics