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

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 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

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

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

OBJECTIVETo observe the effect of Shenluotong Decoction (SD) on serum levels of aldosterone, monocyte chemoattractant protein-1 (MCP-1), α-smooth muscle protein (α-SMA), and nuclear factor-KB (NF-κB) in obstructive nephropathy rats, and to explore the initial mechanism of SD for inhibiting renal interstitial fibrosis.

METHODSTotally 48 healthy Wistar rats were randomly divided into the sham-operation group (n =12) and the model group (n =36). Renal interstitial fibrosis rat model was established by unilateral ureteral obstruction (UUO). After successful modeling, 36 rats were randomly divided into the model group, the Chinese medicine group, and the Western medicine group, 12 in each group. Eplerenone was added in the forage at the daily dose of 100 mg/kg for rats in the Western medicine group. Chinese medicine was added in the forage at the daily dose of 26 g/kg for rats in the Chinese medicine group. Equal volume of normal saline was administered to rats in the sham-operation group and the model group. All medication was performed once daily. The obstructive kidneys were extracted ten days after medication. The pathomorphological changes were observed. The contents of serum aldosterone and MCP-1, and the protein or mRNA expression of MCP-1, α-SMA, and NF-KB were detected.

RESULTSCompared with the sham-operation group, infiltration of a large amount of inflammatory cells and collagen deposition significantly increased, serum contents of aldosterone and MCP-1 obviously increased (P < 0.01), the expression of MCP-1 mRNA and protein were significantly up-regulated (P <0.01), the protein expression of α-SMA and NF-KB were significantly enhanced in the model group (P <0.01). Com- pared with the model group, infiltration of inflammatory cells and renal collagen deposition were attenua- ted in the Chinese medicine group and the Western medicine group, the serum MCP-1 level were reduced, and the mRNA and protein expression of MCP-1 were significantly down-regulated (P <0.01), the protein expression of α-SMA and NF-KB were obviously inhibited (P <0. 01). At the same time, serum aldosterone level was reduced in the Chinese medicine group (P <0.01).

CONCLUSIONSinflammatory lesions of the renal tissue could promote the progress of interstitial fibrosis in rats with obstructive nephropathy. SD could attenuate interstitial fibrosis through reducing serum contents of aldosterone and MCP-1, down-regulating MCP-1/ NF-KB, and inhibiting the expression of α-SMA.

Animals ; Chemokine CCL2 ; drug effects ; metabolism ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fibrosis ; Kidney ; drug effects ; Kidney Diseases ; drug therapy ; genetics ; NF-kappa B ; drug effects ; metabolism ; RNA, Messenger ; biosynthesis ; Rats, Sprague-Dawley ; Ureteral Obstruction ; drug therapy ; genetics

OBJECTIVETo investigate effects of supplementation of folate on the expression of monocyte chemoattractant protein-1 (MCP-1) in aortic endothelium and release from peripheral blood mononuclear cells (PBMC) in rats with hyperhomocysteinemia induced by ingestion of excess methionine.

METHODSThirty male SD rats were randomly divided into 3 groups (n = 10 for each group): control group (Control), high homocysteinemia group (Hhcy), and folate supplementation group (FA). They were fed with nomal diet, normal diet enriched by 1.7% methionine, and normal diet plus 1.7% methionine and 0.006% folate, respectively, for 45 days. The levels of total plasma homocysteine (Thcy) were measured by high performance liquid chromatography and the concentrations of chemokine MCP-1 released from PBMC stimulated by oxidized low density lipoprotein were detected by enzyme immunoassays. The expression of MCP-1 on aortas of rats was detected by immunohistochemistry and Western blot.

RESULTSA high methionine diet for 45 days induced hyperhomocysteinemia. Folate supplementation to high-methionine diet significantly decreased plasma Thcy levels (P < 0.01). The expression of MCP-1 in aortic endothelium and the levels of MCP-1 released from PBMC stimulated by oxidized low density lipoprotein were significantly higher in rats of Hhcy group than in rats of control group (P < 0.05, P < 0.01). During supplementation of folate, normalization of Thcy levels was accompanied by a marked reduction of MCP-1 expression in aortic endothelium and by a significant decrease of MCP-1 released from PBMC stimulated by oxidized low density lipoprotein (P < 0.05, P < 0.01).

CONCLUSIONFolate supplementation can prevent an elevation of homocysteine levels in the blood and decrease the expression MCP-1 in aortic endothelium and release of MCP-1 from PBMC in rats with hyperhomocysteinemia.

Animals ; Aorta ; cytology ; metabolism ; Chemokine CCL2 ; biosynthesis ; genetics ; Endothelium, Vascular ; metabolism ; Folic Acid ; pharmacology ; Homocysteine ; blood ; Hyperhomocysteinemia ; blood ; chemically induced ; metabolism ; Leukocytes, Mononuclear ; metabolism ; Male ; Methionine ; Random Allocation ; Rats ; Rats, Sprague-Dawley