OBJECTIVETo study the effect of oral administration of dimethyl dimethoxy biphenyl dicarboxylate (DDB) on adjusting angiogeneic/inflammatory mediators and ameliorating the pathology of bones in rats with collagen-induced arthritis (CIA).

METHODSWistar rat model of CIA was set up using bovine collagen type II. Fifty rats were divided into five groups randomly: normal, CIA model, DDB treatment, methotrexate (MTX) treatment, and combined DDB+MTX treatment. Ankle joints of rats were imaged with digital X-ray machine to show the destruction of joints. Fore and hind paw and knee joints were removed above the ankle joint then processed for haematoxylin and eosin staining. Plasma levels of vascular endothelial growth factor (VEGF), platelet derived growth factor, interleukin-8 (IL-8), IL-4, tumor necrosis factor α (TNF-α), and cyclooxygenase-2 (COX-2) were quantified by enzyme-linked immunosorbent assay. Nitric oxide levels were detected by Griess reagent.

RESULTSCompared with the CIA model group, a remarkable reduction in various angiogenic (VEGF and IL-8) and inflammatory mediators (TNF-α, IL-4 and COX-2) after treatment with DDB either alone or combined with MTX P<0.05 or P<0.01). Histopathological and X-ray findings were confirmatory to the observed DDB anti-arthritic effect. The DDB-treated group showed amelioration in signs of arthritis which appeared essentially similar to normal.

CONCLUSIONOur data shed light on the therapeutic efficacy of DDB in experimental rheumatoid arthritis (RA) compared with a choice drug (MTX) and it may be offered as a second-line drug in the treatment of RA.

Animals ; Arthritis, Experimental ; chemically induced ; diagnostic imaging ; drug therapy ; pathology ; Arthritis, Rheumatoid ; diagnostic imaging ; drug therapy ; pathology ; Collagen ; Cyclooxygenase 2 ; blood ; Dioxoles ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Female ; Interleukin-4 ; blood ; Interleukin-8 ; blood ; Methotrexate ; therapeutic use ; Nitric Oxide ; biosynthesis ; Platelet-Derived Growth Factor ; analysis ; Radiography ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood ; Vascular Endothelial Growth Factor A ; blood

Myeloid-derived suppressor cells (MDSCs) play a crucial role in T cell dysfunction, which is related to poor outcome in patients with severe trauma. Cyclooxygenase-2 (Cox-2) contributes to immune disorder in trauma and infection via production of prostaglandin E2. However, the role of Cox-2 in the accumulation and function of MDSCs after traumatic stress has not been fully elucidated. In the present study, we treated murine trauma model with NS398, a selective Cox-2 inhibitor. Then the percentages of CD11b+/Gr-1+ cells, proliferation and apoptosis of CD4+ T cells were determined. Arginase activity and arginase-1 (Arg-1) protein expression of splenic CD11b+/Gr-1+ cells, and delayed-type hypersensitivity (DTH) response were analyzed. The results showed that Cox-2 blockade significantly decreased the percentages of CD11b+/Gr-1+ cells in the spleen and bone marrow 48 and 72 h after traumatic stress. NS398 inhibited arginase activity and down-regulated the Arg-1 expression of splenic CD11b+/Gr-1+ cells. Moreover, NS398 could promote proliferation and inhibit apoptosis of CD4+ T cells. It also restored DTH response of traumatic mice. Taken together, our data revealed that Cox-2 might play a pivotal role in the accumulation and function of MDSC after traumatic stress.
Animals ; Apoptosis ; drug effects ; Arginase ; biosynthesis ; CD11b Antigen ; biosynthesis ; CD4-Positive T-Lymphocytes ; drug effects ; metabolism ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; biosynthesis ; Cyclooxygenase 2 Inhibitors ; administration & dosage ; Gene Expression Regulation ; drug effects ; Humans ; Mice ; Myeloid Progenitor Cells ; metabolism ; pathology ; Nitrobenzenes ; administration & dosage ; Stress Disorders, Traumatic ; drug therapy ; genetics ; pathology ; Sulfonamides ; administration & dosage

OBJECTIVE: To investigate the significance of expression of COX-2, p21, Ki67 and HPV in nasal inverted papilloma. METHOD: Detecting COX-2, p21, Ki-67 in 30 cases of nasal inverted papilloma (NIP), 20 cases of nasal polyps (NP) and 10 cases of normal nasal mucosa (NM) by two step immunohistochemical method, and HPV virus by flow-through hybridization method. RESULT: The positive expression rate of COX-2 and Ki-67 in NIP, NP and NM group was decreased in turn, COX-2 had significant difference in the groups(χ2 = 30.00, P< 0. 05); the positive expression rate of Ki-67 had significant differences between NIP and NM group (χ2 = 8. 533, P<0. 05). The expression of COX-2 in NIP tissues was positively correlate with that of Ki-67 by using Spearman rank correlation analysis (r=0.78, P<0.05). Expression of p21 were not observed in NIP group. The positive rate of HPV was 26. 67% in 30 cases of NIP, all of HPV16 type. CONCLUSION COX-2, Ki-67 and HPV infection have certain correlation with the occurrence of NIP. The occurrence of NIP has relationship with inflammatory reaction mediated by COX-2. Ki-67 can well reflect the proliferation activity of tumor cells, and can be used to measure the proliferation rate of nasal inverted papilloma. The COX-2 and Ki-67 have a synergistic role in the pathogenesis of NIP. p21 has no significant relationship with the incidence of NIP. HPV infection is related to the pathogenesis of NIP, but not as a;major factor in the pathogenesis of NIP.
Case-Control Studies ; Cyclin-Dependent Kinase Inhibitor p21 ; biosynthesis ; Cyclooxygenase 2 ; biosynthesis ; Humans ; Ki-67 Antigen ; biosynthesis ; Nasal Mucosa ; Nasal Polyps ; Nose Neoplasms ; genetics ; virology ; Papilloma, Inverted ; genetics ; virology ; Papillomavirus Infections

OBJECTIVEBuckwheat has been considered as a potential source of nutraceutical components on the world market of probiotic foodstuffs. The purpose of this study was to evaluate the effects of tartary buckwheat (Fagopyrum tataricum) sprouts on oxidation and pro-inflammatory mediators.

METHODSThe anti-oxidant effects of buckwheat extract (BWE) and rutin were evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH)- and nitric oxide (NO)-scavenging activities, serum peroxidation and chelating assays. Lipopolysaccharide (LPS)-stimulated RAW264.7 cells were used to evaluate anti-inflammatory activities of buckwheat and rutin. NO production in LPS-stimulated RAW264.7 cells was determined by using Griess reagent. The expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-κB) p65 subunit in cytosolic and nuclear portions were determined by Western blot analysis. Also, the production of inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) was determined by enzyme-linked immunosorbent assay.

RESULTSInhibitory concentration 50 values for DPPH- and NO-scavenging activities of BWE were 24.97 and 72.54 μg/mL respectively. BWE inhibited serum oxidation and possessed chelating activity. Furthermore, BWE inhibited IL-6 and TNF-α production in LPS-stimulated RAW264.7 cells. Also, BWE inhibited iNOS and COX-2 expression and NF-κB p65 translocation.

CONCLUSIONBuckwheat sprouts possessed strong antioxidant activity and inhibited production of pro-inflammatory mediators in the applied model systems. Thus, buckwheat can be suggested to be beneficial in inflammatory diseases by inhibiting the free radicals and inflammatory mediators.

Animals ; Cells, Cultured ; Cyclooxygenase 2 ; analysis ; Fagopyrum ; Free Radical Scavengers ; pharmacology ; Inflammation Mediators ; antagonists & inhibitors ; Interleukin-6 ; biosynthesis ; Lipopolysaccharides ; pharmacology ; Macrophages ; drug effects ; metabolism ; Mice ; NF-kappa B ; metabolism ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; analysis ; Plant Extracts ; pharmacology ; Tumor Necrosis Factor-alpha ; biosynthesis

BACKGROUNDMechanical stress plays an important role in the maintenance of bone homeostasis. Current hypotheses suggest that interstitial fluid flow is an important component of the system by which tissue level strains are amplified in bone. This study aimed to test the hypothesis that the short-term and appropriate fluid shear stress (FSS) is expected to promote the terminal differentiation of pre-osteoblasts and detect the expression profile of microRNAs in the FSS-induced osteogenic differentiation in MC3T3-E1 cells.

METHODSMC3T3-E1 cells were subjected to 1 hour of FSS at 12 dyn/cm(2) using a parallel plate flow system. After FSS treatment, cytoskeleton immunohistochemical staining and microRNAs (miRNAs) were detected immediately. Osteogenic gene expression and immunohistochemical staining for collagen type I were tested at the 24th hour after treatment, alkaline phosphatase (ALP) activity assay was performed at 24th, 48th, and 72 th hours after FSS treatment, and Alizarin Red Staining was checked at day 12.

RESULTSOne hour of FSS at 12 dyn/cm(2) induced actin stress fiber formation and rearrangement, up-regulated osteogenic gene expression, increased ALP activity, promoted synthesis and secretion of type I collagen, enhanced nodule formation, and promoted terminal differentiation in MC3T3-E1 cells. During osteogenic differentiation, expression levels of miR-20a, -21, -19b, -34a, -34c, -140, and -200b in FSS-induced cells were significantly down-regulated.

CONCLUSIONThe short-term and appropriate FSS is sufficient to promote terminal differentiation of pre-osteoblasts and a group of miRNAs may be involved in FSS-induced pre-osteoblast differentiation.

Actins ; chemistry ; Alkaline Phosphatase ; metabolism ; Animals ; Cell Differentiation ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Core Binding Factor Alpha 1 Subunit ; genetics ; Cyclooxygenase 2 ; genetics ; Gene Expression Profiling ; Mice ; MicroRNAs ; physiology ; Osteoblasts ; cytology ; Osteogenesis ; Stress, Mechanical ; Stress, Physiological

The effect of extracellular beta-(1-->3), (1-->6)-glucan, produced by Paenibacillus polymyxa JB115, on nitric oxide (NO) production in RAW264.7 macrophages was investigated. beta-glucan induced the production of NO by RAW264.7 macrophages in a concentration- and time-dependent manner. Moreover, beta-glucan stimulation increased the mRNA expression of iNOS, COX-2 and IL-6 in RAW264.7 macrophages in a concentration-dependent manner.
Animals ; Bacillus/*metabolism ; Cell Line ; Cyclooxygenase 2/biosynthesis/genetics ; Interleukin-6/biosynthesis/genetics ; Lipopolysaccharides/pharmacology ; Macrophages/*drug effects/enzymology/immunology ; Mice ; Nitric Oxide/*biosynthesis/immunology ; Nitric Oxide Synthase Type II/biosynthesis/genetics/metabolism ; RNA, Messenger/biosynthesis/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; beta-Glucans/metabolism/*pharmacology

OBJECTIVETo investigate the role of P38 signaling pathway in modulating the expression of cyclooxygenase-2 (COX-2) in the substantia nigra (SN) of mice with 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced Parkinson disease (PD), and explore the possible mechanism of the dopaminergic (DA) neuron death in PD and the effects of ginsenoside Rg1 on the P38 signaling pathway and DA neurons.

METHODSC57BL6 mice were treated with MPTP to produce the subacute PD model, and the behavioral changes were observed. Immunohistochemistry and Western blotting for tyrosine hydroxylase (TH), COX-2, prostaglandin E2 (PGE2) and phosphorylated P38 (p-P38) were used to observe the changes of positive cell number in the midbrain after treatment with ginsenoside Rg1.

RESULTSCompared with the control mice, the mice with PD presented with typical symptoms of PD. The number of p-P38-, COX-2-, and PGE2-positive cells significantly increased in the SN area 6 h after the 3rd injection of 30 mg/kg MPTP (P<0.01). The number of TH-positive neurons in the PD model group was substantially reduced by about 60% (P<0.01) in 24 h after the 5th injection of MPTP. In mice with ginsenoside Rg1 treatment, the number of p-P38-, COX-2-, and PGE2-positive cells was reduced obviously 6 h after the 3rd injection of MPTP as compared with that in the model group (P<0.01). The number of TH-positive neurons in the SN was decreased by only 30% (P<0.01 vs control group) 24h after the 5th injection of MPTP.

CONCLUSIONP38 signaling pathway may play an important role in modulating COX-2 expression in the SN in the early stage of MPTP-induced subacute PD, and ginsenoside Rg1 may act on the P38 signaling pathway to protect the DA neurons in PD.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ; Animals ; Blotting, Western ; Cyclooxygenase 2 ; biosynthesis ; Ginsenosides ; pharmacology ; Immunohistochemistry ; Male ; Mice ; Mice, Inbred C57BL ; Neurons ; drug effects ; metabolism ; pathology ; Parkinson Disease, Secondary ; chemically induced ; metabolism ; physiopathology ; Signal Transduction ; drug effects ; Substantia Nigra ; drug effects ; metabolism ; pathology ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo determine the levels of MMP-2 and COX-2 mRNA in bladder transitional cell carcinoma tissues and explore their relationship.

METHODSWe enrolled in this study 42 patients with bladder transitional cell carcinoma, including Ta-T1 (n = 18), T2-T4 (n = 24), G1 (n = 12), G2 (n = 19), G3 (n = 11), metastasis (n =26) and non-metastasis (n = 16). Another 5 cases of normal bladder tissues were taken as controls, and the levels of MMP-2 and COX-2 mRNA were detected by RT-PCR.

RESULTSThe relative expressions of COX-2 mRNA were 1.038 +/- 0. 484 in Ta-T1, 1.489 +/- 0.584 in T2-T4, 0.920 +/- 0.442 in G1, 1.338 +/- 0.584 in G2 and 1.632 +/- 0.515 in G3, all significantly higher than that of the controls (0.460 +/- 0.224, P < 0.05). And the corresponding relative levels of MMP-2 mRNA were 1.107 +/- 0.384, 1.604 +/- 0.425, 0.971 +/- 0.370, 1.445 +/- 0.378 and 1.755 +/- 0.387, also significantly higher than that of the latter group (0.423 +/- 0.227, P < 0.05). The COX-2 and MMP-2 mRNA levels in the tumor tissues with and without metastasis were 1.591 +/- 0.455 vs 0.815 +/- 0.430 and 1.676 +/- 0.339 vs 0.927 +/- 0.228, (P < 0.01), respectively, with a positive correlation between the mRNA level of COX-2 and that of MMP-2 (r = 0. 703, P < 0.01).

CONCLUSIONMMP-2 and COX-2 mRNA are highly expressed in bladder transitional cell carcinoma tissues and their expressions are positively correlated with the degree of malignancy. MMP-2 and COX-2 might play a synergetic role in the pathogenesis and progression of bladder transitional cell carcinoma.

Adult ; Aged ; Carcinoma, Transitional Cell ; metabolism ; pathology ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Female ; Humans ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; genetics ; Urinary Bladder Neoplasms ; metabolism ; pathology

OBJECTIVETo study the effect of Xiaojin Wan on the expression of COX-2 in the prostate tissues of rats with prostatitis pain, and the action mechanism of the drug alleviating the prostatitis pain.

METHODSSixty male Wistar rats were randomized into two groups, 10 as blank controls, injected with aqua pro injection into the ventral part of prostate, and the other 50 as prostatitis pain models, given complete Freund's adjuvant (CFA). Three days later, the pain model rats were again equally divided into 5 subgroups: model control, Celecoxib Capsules, high-, median- and low-dose Xiaojin Wan, receiving intragastric administration of distilled water, Celecoxib Capsules and different doses of Xiaojin Wan respectively for 4 weeks. Then they were killed, the harvested tissues fixed with 10% paraformaldehyde and the changes of the COX-2 expression in the prostate detected with the immunohistochemical technique and graphics video analysis system.

RESULTSThe expression of COX-2 was strong in the model group, significantly lower in the high- and median-dose and the Celecoxib Capsules groups than in the model control (P < 0.01) as well as in the high-dose than in the median- and low-dose groups (P < 0.01).

CONCLUSIONXiaojin Wan may alleviate prostatitis pain by inhibiting the expression of COX-2 in prostate tissues.

Animals ; Chronic Disease ; Cyclooxygenase 2 ; biosynthesis ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Immunohistochemistry ; Male ; Pain ; drug therapy ; enzymology ; etiology ; Phytotherapy ; Prostate ; drug effects ; enzymology ; pathology ; Prostatitis ; complications ; drug therapy ; enzymology ; Random Allocation ; Rats ; Rats, Wistar

Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) is a tumor suppressor. Although it is well known to have various physiological roles in cancer, its inhibitory effect on inflammation remains poorly understood. In the present study, a human PTEN gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-PTEN fusion protein. The expressed and purified PEP-1-PTEN fusion protein were transduced efficiently into macrophage Raw 264.7 cells in a time- and dose- dependent manner when added exogenously in culture media. Once inside the cells, the transduced PEP-1-PTEN protein was stable for 24 h. Transduced PEP-1-PTEN fusion protein inhibited the LPS-induced cyclooxygenase 2 (COX-2) and iNOS expression levels in a dose-dependent manner. Furthermore, transduced PEP-1-PTEN fusion protein inhibited the activation of NF-kappa B induced by LPS. These results suggest that the PEP-1-PTEN fusion protein can be used in protein therapy for inflammatory disorders.
Animals ; Cell Line ; Cyclooxygenase 2/*metabolism ; Cysteamine/*analogs & derivatives ; Enzyme Activation ; Humans ; Lipopolysaccharides/*pharmacology ; Macrophages/*metabolism ; Mice ; NF-kappa B/metabolism ; Nitric Oxide/*biosynthesis ; Nitric Oxide Synthase Type II/metabolism ; PTEN Phosphohydrolase/*genetics ; Peptides/*genetics ; Recombinant Fusion Proteins/*biosynthesis/genetics ; Signal Transduction