This experiment was designed to investigate the influence of two biomaterials, titanium oxide (Ti-O) and stainless steel (SS), on the cytokine expression of macrophage, and further, to evaluate their biocompatibility. After being co-cultured with Ti-O and SS for 72 h, the cell number and morphology of macrophages attached on materials were detected by fluorescent microscope and SEM. Nitride oxide (NO) and monocyte chemoattractant protein 1 (MCP-1) released by the macrophages co-cultured with different materials were also examined and compared. We found that the cell number of macrophages attached to Ti-O was smaller than that attached to SS. The levels of NO and MCP-1 released by the macrophages co-cultured with Ti-O were lower when compared with those released by macrophages co-cultured with SS. These results demonstrate that Ti-O has better biocompatibility than does SS.
Animals ; Biocompatible Materials ; pharmacology ; Cells, Cultured ; Chemokine CCL2 ; metabolism ; Macrophages ; cytology ; metabolism ; Male ; Materials Testing ; methods ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stainless Steel ; pharmacology ; Titanium ; pharmacology

OBJECTIVETo investigate the effect of exendin-4 on the expression of monocyte chemoattractant protein-1 (MCP-1) and fibronectin (FN) in rat glomerular mesangial cells in vitro.

METHODSRat glomerular mesangial cells were divided into 5 groups, namely control group, tumor necrosis factor-α (TNF-α) group (10 ng/ml), TNF-α (10 ng/ml)+E1 (1 nmol/L exendin-4) group, TNF-α (10 ng/ml)+E5 (5 nmol/L exendin-4) group, and TNF-α (10 ng/ml)+E10 (10 nmol/L exendin-4) group. After cultured 24 h or 48 h, RNA were extracted to determine the expression of MCP-1 with real-time PCR, the supernatant were collected to determine the expression of MCP-1 and FN with ELISA.

RESULTSCompared with control group, the cells treated with TNF-α for 24 h showed significantly increase the expression of MCP-1 and FN (P<0.01), exendin-4 significantly reduced the expression of MCP-1 and FN in TNF-α+E5 group and TNF-α+E10 group (P<0.05). After 48h incubation, the expression of MCP-1 and FN increased significantly in TNF-α group (P<0.01), which was lowered by exendin-4 in TNF-α+E1,TNF-α+E5 and TNF-α+E10 groups (P<0.05).

CONCLUSIONExendin-4 has an intrinsic capability to concentration- and time-dependently inhibit TNF-α-induced expression of MCP-1 and FN in rat mesangial cells, suggesting the beneficial effect of exendin-4 in preventing and treating diabetic nephropathy.

Animals ; Cells, Cultured ; Chemokine CCL2 ; metabolism ; Diabetic Nephropathies ; metabolism ; Glomerular Mesangium ; cytology ; Mesangial Cells ; drug effects ; metabolism ; Peptides ; pharmacology ; Rats ; Tumor Necrosis Factor-alpha ; pharmacology ; Venoms ; pharmacology

OBJECTIVETo investigate the effect of cryptoporus polysaccharide(CP)on lipopolysaccharide(LPS)-induced production of monocyte chemoattractant protein-1(MCP-1)in human lung epithelial A549 cells.

METHODSA549 cells were stimulated with LPS in the presence or absence of CP. The protein concentration and mRNA expression of MCP-1 were determined by enzyme-linked-immunosobent assay(ELISA)and semi-quantitative RT-PCR, respectively.

RESULTThe protein concentration of MCP-1 was significantly increased by LPS 1000 microg/L at 24 h. There were no effects on the growth and viability of A549 cells in the presence of CP 100 microg/L or dexamethasone 1 mumol/L. However, CP 100 microg/L or dexamethasone 1 micromol/L significantly inhibited the protein concentration and mRNA expression of MCP-1 induced by LPS.

CONCLUSIONCP can regulate MCP-1 production, which may be associated with its effects on lung inflammation.

Cell Line ; Chemokine CCL2 ; genetics ; metabolism ; Epithelial Cells ; cytology ; metabolism ; Humans ; Lipopolysaccharides ; pharmacology ; Polyporaceae ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Pulmonary Alveoli ; cytology ; RNA, Messenger ; metabolism

OBJECTIVETo investigate the role of protein kinase C (PKC) in thrombin-induced monocyte chemoattractant protein-1(MCP-1) release by human lung fibroblasts (HLF-1).

METHODSCultured human lung fibroblasts HLF-1 were incubated with different concentrations of PKC inhibitors before by thrombin stimulation. MCP-l protein levels in the supernatants were assessed using ELISA, and MCP-1 mRNA levels in the cell lysate were measured by quantitative real-time PCR.

RESULTSThe broad spectrum PKC inhibitors bisindolylmaleimide I and RO-31-8220 obviously inhibited thrombin-induced MCP-l mRNA and protein expressions in HLF-1 cells, whereas Ca(2+)-dependent PKC inhibitor Go 6976 had no such effects.

CONCLUSIONCa(2+)-independent PKC mediates thrombin-induced MCP-1 release in cultured HLF-1 cells.

Cell Line ; Cells, Cultured ; Chemokine CCL2 ; metabolism ; Fibroblasts ; metabolism ; Humans ; Indoles ; pharmacology ; Lung ; cytology ; metabolism ; Protein Kinase C ; antagonists & inhibitors ; Thrombin ; pharmacology

OBJECTIVETo observe the effects of different doses of Wenxiao Decoction on the expression of interleukin-6 (IL-6), intercellular adhesion molecule-1 (ICAM-1), and monocyte chemoattractant protein-1 (MCP-1) in experimental atherosclerotic rabbits and to explore the mechanism by which it alleviates atherosclerosis.

METHODSSixty New Zealand rabbits were randomly divided into six groups: a blank group, a model group, a Simvastatin group, and high-, medium-, and low-dosage Wenxiao Decoction groups. Except for those in the blank group, all rabbits were fed with a high-cholesterol diet. Carotid atherosclerosis was established by balloon-induced carotid artery endothelium injury in conjunction with the high-cholesterol diet. After 8 weeks, all animals were euthanized to evaluate levels of IL-6 and ICAM-1 expressions (by enzyme linked immunosorbent assay) and of MCP-1 (by immunohistochemistry staining).

RESULTSThe expressions of IL-6, ICAM-1, and MCP-1 were significantly increased in all groups except the blank group (P<0.05). However, the rabbits in the Wenxiao Decoction groups and the Simvastatin group showed significantly lower levels of IL-6, ICAM-1, and MCP-1 expression than those in the model group (P<0.05). The expressions of IL-6, ICAM-1, and MCP-1 in the highdosage Wenxiao Decoction group and the Simvastatin group were lower than those in the low-dosage Wenxiao Decoction group (P<0.05). The expression of MCP-1 in medium-dosage Wenxiao Decoction group was lower than that in the low-dosage group (P<0.05).

CONCLUSIONSHigh, medium, and low doses of Wenxiao Decoction can inhibit the expressions of IL-6, ICAM-1, and MCP-1, which may prevent and stabilize atherosclerotic plaques. There may be a direct relationship between dosage and therapeutic efficacy of Wenxiao Decoction.

Animals ; Atherosclerosis ; chemically induced ; metabolism ; pathology ; Chemokine CCL2 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-6 ; metabolism ; Rabbits

OBJECTIVETo investigate the effects of very low-density lipoprotein (VLDL) on cellular lipid accumulation and the expression of monocyte chemoattractant protein-1 (MCP-1) in human mesangial cells.

METHODSAn established stable human mesangial cell line (HMCL) was used in all experiments. VLDL-induced cellular lipid deposition was visualized by Oil Red O staining and analyzed quantitatively by standard enzymatic procedures. MCP-1 mRNA and protein expression levels in treated HMCLs were determined by real-time quantitative RT-PCR and enzyme-linked immunosorbent assay, respectively. For adhesion study, HMCLs were treated with VLDL for 12 hours, followed by a one-hour incubation with THP-1 cells.

RESULTSVLDL induced cellular lipid accumulation in HMCLs in a time- (0-24 h) and dose- (0-200 microg/ml) dependent manner, and the principal component of accumulated lipid is triglyceride. In HMCLs, MCP-1 mRNA expression was promoted by VLDL in a time- (0-6 h) and dose- (0-100 microg/ml) dependent manner, and VLDL also enhanced MCP-1 secretion in a dose-dependent manner. Such an effect was accompanied by increased adhesion of monocytes to HMCLs.

CONCLUSIONSVLDL can induce cellular triglyceride accumulation and upregulate the expression of MCP-1 in human mesangial cells. Hence, VLDL may be involved in the pathogenesis of lipid-mediated renal injury.

Cell Line ; Chemokine CCL2 ; genetics ; metabolism ; Humans ; Lipoproteins, VLDL ; pharmacology ; toxicity ; Mesangial Cells ; cytology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Triglycerides ; metabolism

OBJECTIVETo observe the effect of nitrotyrosine on renal expressions of nuclear factor-κB (NF-κB), monocyte chemoattractant protein-1 (MCP-1), and transforming growth factor-β1 (TGF-β1) in rats with diabetic nephropathy (DN).

METHODSRat DN models established by a single intraperitoneal injection of streptozotocin (STZ) were randomly allocated into model group, nitrotyrosine group and ebselen group, with untreated rats as the normal control group. The rats were given the corresponding drugs for 8 weeks, and after the last administration, the 24-h urinary protein level was measured and the kidneys of the rats were harvested for detecting the protein and mRNA expressions of NF-κB, MCP-1 and TGF-β1 with immunohistochemistry and RT-PCR, respectively. The pathological changes of the kidneys were assessed microscopically.

RESULTSCompared with those in the model group, the 24-h urinary protein level and expressions of NF-κB, MCP-1 and TGF-β1 mRNA and protein in the renal tissues were significantly increased by nitrotyrosine treatment, which also caused worsened renal pathology, while treatment with ebselen significantly ameliorated these changes.

CONCLUSIONNitrotyrosine can up-regulate the mRNA and protein expressions of NF-κB, MCP-1 and TGF-β1 and aggravate the inflammatory reaction in the renal tissue of DN rats to promote the progression of DN.

Animals ; Chemokine CCL2 ; metabolism ; Diabetes Mellitus, Experimental ; Diabetic Nephropathies ; metabolism ; Male ; NF-kappa B ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; Tyrosine ; analogs & derivatives ; pharmacology

OBJECTIVETo evaluate the effect and possible mechanism of gypenoside (GP) on expression of inflammatory factors in aortic lesion of rats with high-fat induced atherosclerosis.

METHODSAtherosclerotic rat model was established by feeding high-fat diet and intraperitoneal injection of vitamin D3. Sixty healthy male SD rats were randomly divided into the normal group, the model group, the simvastatin treated group and the three GP groups treated respectively with different dosages of GP. Rats were sacrificed 7 weeks later, their histopathological changes in thoracic aorta were observed by light microscope; expressions of intercellular adhesion molecule 1 (ICAM-1), monocyte chemotactic protein-1 (MCP-1) and nuclear factor-kappaBp65 (NF-kappaBp65) in aortic wall were detected by immunohistochemistry; serum level of oxidized low-density lipoprotein (ox-LDL) was determined by ELISA; serum total antioxidant capacity determined by colorimetry, and serum malondialdehyde (MDA) level determined by Thiobarbituric acid method.

RESULTSIn comparing with the model group, GPS showed actions in lessening the atherosclerosis lesion; reducing expressions of ICAM-1, MCP-1 and NF-kappaBp65 in aortic wall (P<0.01) and serum levels of MDA, ox-LDL (P < 0.01), as well as increasing the serum level of total antioxidant capacity (P < 0.01 ).

CONCLUSIONGP can down-regulate the expressions of ICAM-1 and MCP-1, inhibit the atherosclerosis formation in experimental rats, its mechanism might be related with its anti-oxidation effect and further inhibiting on the NF-kappaB activation.

Animals ; Atherosclerosis ; metabolism ; pathology ; Chemokine CCL2 ; metabolism ; Gynostemma ; Inflammation ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; NF-kappa B ; metabolism ; Oxidative Stress ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of losartan on the expression of monocyte chemoattractant protein-1 (MCP1) and transforming growth factor-β(1) (TGF-β(1)) in the kidney of rats with unilateral urethral obstruction (UUO) and evaluate protective effect of losartan against reanal interstitial fibrosis.

METHODSRat models of UUO were treated with losartan at the routine dose, high dose, and very high dose (50, 200, and 500 mg/kg daily, respectively), and saline was given to UUO model rats and rats with sham operation. At 7, 14, and 21 days, the tail cuff blood pressure (TCP), 24-h urine protein (Upro), serum Scr, BUN, K(+), percentage of renal damage and renal interstitial fibrosis (%INT) were measured in the rats. MCP1 protein in the renal tissues was detected using immunohistochemistry, and MCP1 and TGF-β(1) mRNA expressions were assayed using RT-PCR.

RESULTSAs the UUO prolonged, Upro, TCP, tubular damage, %INT, and MCP1 and TGF-β(1) mRNA expressions all increased significantly (P<0.05). High and very high doses of losartan, compared with the routine dose, obviously reversed these changes.

CONCLUSIONHigh-dose losartan can effectively control blood pressure, reduce renal damage and fibrosis, and inhibit MCP1 and TGF-β(1) expression in rats with UUO, and at a very high dose, losartan can more effectively reduce 24-h Upro than the high-dose group. High and very high doses of losartan offer better protective effect on the kidney in rats with UUO.

Animals ; Chemokine CCL2 ; metabolism ; Fibrosis ; etiology ; prevention & control ; Kidney ; metabolism ; pathology ; Losartan ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; Ureteral Obstruction ; complications ; drug therapy

OBJECTIVETo explore the effects and related mechanism of nifedipine on vascular inflammation induced by cuff placement.

METHODSAdult male C57BL/6J mice (10 to 12 weeks of age) were assigned to control (no cuff placement without nifedipine), cuff placement (cuff placement without nifedipine) and treatment (cuff placement with nifedipine 1 or 5 mg×kg(-1)×d(-1)) groups. Activity of NF-κB in injured artery was measured 5 days after operation. MCP-1 expression and nuclear translocation of NF-κB were examined in injured artery 7 days after operation.

RESULTSDNA-binding activity of NF-κB was significantly increased in the injured artery 5 days after cuff placement which could be downregulated by nifedipine 5 mg×kg(-1)×d(-1). MCP-1 mRNA expression in the injured arteries was increased 7 days after cuff placement and which could be significantly attenuated by nifedipine 5 mg×kg(-1)×d(-1). Cuff placement decreased the cytoplasmic level of p50, IκBα, IκBβ, and increased the nuclear level of p50. Nifedipine 5 mg×kg(-1)×d(-1) significantly attenuated these changes.

CONCLUSIONOur results suggest that high dose nifedipine could suppresses expression of MCP-1 induced by injured arteries via the inhibin NF-κB DNA binding activity, thereby attenuating vascular inflammation.

Animals ; Blood Vessels ; metabolism ; Chemokine CCL2 ; metabolism ; Inflammation ; Male ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; metabolism ; Nifedipine ; pharmacology ; Vascular Diseases ; metabolism