OBJECTIVETo establish the model of cellular inflammatory responses of gingival epithelial cells in vitro induced by Porphyromonas gingivalis vesicle and to probe into the pathogenesis of Porphyromonas gingivalis in periodontitis.

METHODSThe effect of Porphyromonas gingivalis vesicle on prostaglandin E(2) (PGE(2)) production of gingival epithelial cells was detected by ELISA and the effects of Porphyromonas gingivalis vesicle on cyclooxygenase-2 (COX-2), interleukin (IL)-6 and IL-8 mRNA expression in gingival epithelial cells were determined by Real-time reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSPorphyromonas gingivalis vesicle dose-dependently induced PGE(2) production and up-regulated COX-2, IL-6 and IL-8 mRNA expression in gingival epithelial cells significantly.

CONCLUSIONSCellular inflammatory responses of gingival epithelial cells induced by Porphyromonas gingivalis vesicle may contribute to the initiation and progression of periodontitis.

Bacterial Adhesion ; Cells, Cultured ; Cyclooxygenase 2 ; immunology ; metabolism ; Dinoprostone ; immunology ; metabolism ; Epithelial Cells ; immunology ; metabolism ; microbiology ; Gingiva ; immunology ; metabolism ; microbiology ; Humans ; Interleukin-6 ; immunology ; metabolism ; Interleukin-8 ; immunology ; metabolism ; Porphyromonas gingivalis ; immunology ; pathogenicity

OBJECTIVETo investigate the expression of cyclooxygenase-2(COX-2) and CEA in the tissues adjacent to the tumor within different distances.

METHODSA total of 42 colorectal cancer tissues were collected.The adjacent tissues within 3 cm to the tumor were procured every 1 cm. Normal tissue was also collected. RNA was extracted and the expression of CEA and COX-2 was detected by RT-PCR.

RESULTSThe CEA mRNA levels of the tumor, the tissues of every 1 cm adjacent to the tumor, and the normal tissue were 135.2 ± 23.3, 78.2 ± 17.3, 75.9 ± 16.5, 56.2 ± 10.7, 52.3 ± 12.8, 18.2 ± 7.9, 16.2 ± 6.5, and 16.6 ± 7.0. The levels of COX-2 mRNA in above positions were 134.9 ± 31.1, 79.2 ± 20.2, 77.0 ± 20.5, 62.7 ± 21.9, 58.0 ± 18.1, 21.2 ± 10.3, 18.3 ± 7.6, and 17.1 ± 6.3. These data showed a decreasing trend of CEA and COX-2 as the distance increased from the tumor. The CEA mRNA levels showed positive correlation with the levels of COX-2 mRNA(r=0.725, P<0.01).

CONCLUSIONCEA and COX-2 may be considered to be used as biomarkers for the study of molecular resection margin of colorectal cancer.

Adult ; Aged ; Carcinoembryonic Antigen ; metabolism ; Colorectal Neoplasms ; genetics ; immunology ; pathology ; Cyclooxygenase 2 ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging

BACKGROUND/AIMS: The cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), the proteins that have the role in the gastric carcinogenesis, are stimulated by H. pylori infection in the gastric mucosa. The aim of this study was to evaluate the expression of COX-2 and iNOS proteins one year after the eradication of H. pylori. METHODS: Gastric antral mucosa from fifty eight patients with chronic gastritis who were all infected with H. pylori was examined for the expression of COX-2 and iNOS proteins before and one year after the eradication of H. pylori by immunohistochemical stain. RESULTS: COX-2 and iNOS proteins were expressed in the epithelial cells and interstitial inflammatory cells of gastric mucosa. Percent expressions of COX-2 and iNOS were significantly decreased one year after the eradication in the patients with cured infection, but not in those having persistent H. pylori. COX-2 and iNOS expressions were well correlated with H. pylori density, acute and chronic inflammation of gastric mucosa. CONCLUSIONS: The eradication of H. pylori can decrease the expression of COX-2 and iNOS in the gastric mucosa in long-term period. This seems to be due to the removal of H. pylori itself and related regression of gastric inflammation.
Cyclooxygenase 2/immunology/*metabolism ; Drug Therapy, Combination ; Gastric Mucosa/*enzymology ; Helicobacter Infections/drug therapy ; *Helicobacter pylori ; Humans ; Nitric Oxide Synthase Type II/immunology/*metabolism ; Time Factors

The aim of this study was to investigate the effects of interferon (IFN)-γ on biological characteristics and immunomodulatory property of human umbilical cord-derived mesenchymal stem cells (hUC-MSC). hUC-MSC were treated with IFN-γ 10 ng/ml (IFN-γ group) or without IFN-γ (control group). The phenotype of hUC-MSC was detected by flow cytometry. The proliferation status was detected by CCK-8 method, and its differentiation ability was assessed by oil red O and von Kossa staining. The production of PGE-2 was measured by ELISA, and the mRNA expression levels of COX-2, IDO-1 and IDO-2 in hUC-MSC were detected by real-time quantitative PCR. Furthermore, the proliferation of human peripheral blood mononuclear cells (hPBMNC) was evaluated after co-culture with hUC-MSC, IFN-γ pretreatment or not. The results showed that after IFN-γ stimulation, the expression of SSEA-4 on hUC-MSC decreased significantly [(8.15 ± 2.94) vs (16.42 ± 8.5), P < 0.05], and the expression of CD54 increased [(96.64 ± 3.29) vs (84.12 ± 10.73), P = 0.051]. The immunomodulatory property of hUC-MSC on the proliferation of hPBMNC was enhanced (P < 0.05). All the above mentioned effects were IFN-γ concentration-dependent. When hUC-MSC were stimulated by IFN-γ for 24 h, the production of PGE-2 secreted by hUC-MSC decreased significantly (P < 0.01). The mRNA expression level of COX-2 also decreased though the difference did not reach to statistically significant level. Compared with control group, IDO-1 expression level in IFN-γ group increased significantly (P < 0.01), and the mRNA expression level of IDO-2 remained unchanged. It is concluded that IFN-γ can influence the phenotype of hUC-MSC and enhance the immunomodulatory property of hUC-MSC.
Cells, Cultured ; Cyclooxygenase 2 ; metabolism ; Flow Cytometry ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; metabolism ; Interferon-gamma ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; drug effects ; immunology ; Umbilical Cord ; cytology

This experiment focused on MAPK activation in host cell invasion and replication of T. gondii, as well as the expression of CC chemokines, MCP-1 and MIP-1 alpha , and enzyme, COX-2/prostaglandin E2 (PGE2) in infected cells via western blot, [3H]-uracil incorporation assay, ELISA and RT-PCR. The phosphorylation of ERK1/2 and p38 in infected HeLa cells was detected at 1 hr and/or 6 hr postinfection (PI). Tachyzoite proliferation was reduced by p38 or JNK MAPK inhibitors. MCP-1 secretion was enhanced in infected peritoneal macrophages at 6 hr PI. MIP-1 alpha mRNA was increased in macrophages at 18 hr PI. MCP-1 and MIP-1 alpha were reduced after treatment with inhibitors of ERK1/2 and JNK MAPKs. COX-2 mRNA gradually increased in infected RAW 264.7 cells and the secretion of COX-2 peaked at 6 hr PI. The inhibitor of JNK suppressed COX-2 expression. PGE2 from infected RAW 264.7 cells was increased and synthesis was suppressed by PD98059, SB203580, and SP600125. In this study, the activation of p38, JNK and/or ERK1/2 MAPKs occurred during the invasion and proliferation of T. gondii tachyzoites in HeLa cells. Also, increased secretion and expression of MCP-1, MIP-1 alpha , COX-2 and PGE2 were detected in infected macrophages, and appeared to occur via MAPK signaling pathways.
Toxoplasmosis/*enzymology/*immunology ; Toxoplasma/*immunology/*metabolism ; Mitogen-Activated Protein Kinases/*metabolism ; Mice, Inbred BALB C ; Mice ; Macrophages, Peritoneal/enzymology/immunology/parasitology ; Humans ; Hela Cells ; Enzyme Activation ; Cyclooxygenase 2/*biosynthesis ; Chemokines/*biosynthesis ; Animals

Inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) have been known to be involved in various pathophysiological processes such as inflammation. This study was performed to determine the regulatory function of superoxide dismutase (SOD) on the LPS-induced expression of iNOS, and COX-2 in RAW 264.7 cells. When a cell-permeable SOD, Tat-SOD, was added to the culture medium of RAW 264.7 cells, it rapidly entered the cells in a dose-dependent manner. Treatment of RAW 264.7 cells with Tat-SOD led to decrease in LPS-induced ROS generation. Pretreatment with Tat-SOD significantly inhibited LPS-induced expression of iNOS and NO production but had no effect on the expression of COX-2 and PGE2 production in RAW 264.7 cells. Tat-SOD inhibited LPS-induced NF-kappaB DNA binding activity, IkappaBalpha degradation and activation of MAP kinases. These data suggest that SOD differentially regulate expression of iNOS and COX-2 in LPS-stimulated RAW 264.7 cells.
Animals ; Cell Line ; Cyclooxygenase 2/*genetics/metabolism ; Cytokines/immunology ; *Gene Expression Regulation ; Lipopolysaccharides/immunology/metabolism ; Mice ; Mitogen-Activated Protein Kinase Kinases/metabolism ; NF-kappa B/metabolism ; Nitric Oxide/metabolism ; Nitric Oxide Synthase Type II/*genetics/metabolism ; Reactive Oxygen Species/metabolism ; Superoxide Dismutase/*metabolism

PURPOSE : Cyclooxygenase-2 (COX-2) and its metabolite, PGE2 affect multiple tumorigenesis, including angiogenesis, invasion, and tumor-induced immune suppression. Their overexpression is association with impaired immune cell function in many tumors. Indoleamine 2,3-dioxygenase (IDO) is an emerging immuno-regulatory enzyme that can catalyze the initial rate-limiting step in tryptophan catabolism, by causing tryptophan depletion can block T lymphocyte activation, and thus, enable tumor cells to escape from immune system. Although the potential of immunosuppression associated with tumorproduced COX-2 has been suggested, the mechanism of immunosuppression in tumor immunology is not yet well defined. Thus, we hypothesized that the tumor immunity of COX-2 could be partly due to IDO-dependent immune tolerance. To test this hypothesis, we evaluated IDO expression in cancer cells treated with selective COX-2 inhibitor. MATERIALS AND METHODS : The A549 human adenocarcinoma cell line, murine Lewis lung carcinoma (LLC) cell line and C57Bl/6 mice were used for in vitro and in vivo studies. In vitro studies, A549 cells were treated with various concentrations of COX-2 inhibitor (PTPBS) or PGE2. IDO enzyme activity and protein expression were checked by IDO enzyme activity assay and Western blotting. In vivo study, the 20 mice were randomized into normal control, LLC inoculated control, and low and high selective COX-2 inhibitor (celecoxib 25 or 250 mg/kg/day) treated LLL inoculated mice groups (n=5 per group). At one month, mice were sacrificed and tumor mass was isolated for quantification of IDO expression by immunohistochemical stain and western blotting. RESULTS : In vitro studies, PTPBS treated A549 cells showed a significant decreased in IDO enzyme activity and expression but PGE2 treated A549 cells showed increased in IDO expression. In vivo studies, the tumor mass and lung metastasis were attenuated by celecoxib (respectively, p<0.05, p<0.01). Compared with the LLC inoculated control group, mice treated with celecoxib had significant reductions in IDO expression of tumor mass (IDO immunohistochemical stain and western blotting ). CONCLUSION : The present study reveals that COX-2 inhibitor serves to restore the tumor-induced IDO expression and promotes antitumor reactivity in an immunocompetent murine lung cancer model. These findings further support the suggestion that COX-2 inhibitor is a potential pharmacological immunotherapy in cancer
Adenocarcinoma ; Allergy and Immunology ; Animals ; Blotting, Western ; Carcinogenesis ; Carcinoma, Lewis Lung ; Cell Line ; Cyclooxygenase 2* ; Dinoprostone ; Humans ; Immune System ; Immune Tolerance* ; Immunosuppression ; Immunotherapy ; Indoleamine-Pyrrole 2,3,-Dioxygenase* ; Lung ; Lung Neoplasms ; Lymphocyte Activation ; Metabolism ; Mice ; Neoplasm Metastasis ; Tryptophan ; United Nations ; Celecoxib

Endoplasmic reticulum (ER) stress regulates a wide range of cellular responses including apoptosis, proliferation, inflammation, and differentiation in mammalian cells. In this study, we observed the role of 2-deoxy-D-glucose (2DG) on inflammation of chondrocytes. 2DG is well known as an inducer of ER stress, via inhibition of glycolysis and glycosylation. Treatment of 2DG in chondrocytes considerably induced ER stress in a dose- and time-dependent manner, which was demonstrated by a reduction of glucose regulated protein of 94 kDa (grp94), an ER stress-inducible protein, as determined by a Western blot analysis. In addition, induction of ER stress by 2DG led to the expression of COX-2 protein with an apparent molecular mass of 66-70kDa as compared with the normally expressed 72-74 kDa protein. The suppression of ER stress with salubrinal (Salub), a selective inhibitor of eif2-alpha dephosphorylation, successfully prevented grp94 induction and efficiently recovered 2DG-modified COX-2 molecular mass and COX-2 activity might be associated with COX-2 N-glycosylation. Also, treatment of 2DG increased phosphorylation of Src in chondrocytes. The inhibition of the Src signaling pathway with PP2 (Src tyrosine kinase inhibitor) suppressed grp94 expression and restored COX-2 expression, N-glycosylation, and PGE2 production, as determined by a Western blot analysis and PGE2 assay. Taken together, our results indicate that the ER stress induced by 2DG results in a decrease of the transcription level, the molecular mass, and the activity of COX-2 in rabbit articular chondrocytes via a Src kinase-dependent pathway.
Animals ; Cartilage, Articular/pathology ; Cells, Cultured ; Chondrocytes/drug effects/immunology/*metabolism/pathology ; Cyclooxygenase 2/genetics/*metabolism ; Deoxyglucose/*pharmacology ; Down-Regulation ; Endoplasmic Reticulum/drug effects/*metabolism/pathology ; Glycosylation/drug effects ; Inflammation ; Rabbits ; Signal Transduction/drug effects ; Stress, Physiological/drug effects/immunology ; src-Family Kinases/*metabolism

PURPOSE: Cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 play a key role in the pathogenesis of in-stent restenosis. We investigated the effect of a short-term therapy of celecoxib, a COX-2 inhibitor, with or without doxycycline, an MMP inhibitor, after coronary stenting on inflammatory biomarkers and neointimal hyperplasia. MATERIALS AND METHODS: A total of 75 patients (86 lesions) treated with bare metal stents were randomized into three groups: 1) combination therapy (200 mg celecoxib and 20 mg doxycycline, both twice daily), 2) celecoxib (200 mg twice daily) only, and 3) non-therapy control. Celecoxib and doxycycline were administered for 3 weeks after coronary stenting. The primary endpoint was neointimal volume obstruction by intravascular ultrasound (IVUS) at 6 months. The secondary endpoints included clinical outcomes, angiographic data, and changes in blood levels of inflammatory biomarkers. RESULTS: Follow-up IVUS revealed no significant difference in the neointimal volume obstruction among the three treatment groups. There was no difference in cardiac deaths, myocardial infarctions, target lesion revascularization or stent thrombosis among the groups. Blood levels of high-sensitivity C-reactive protein, soluble CD40 ligand, and MMP-9 varied widely 48 hours and 3 weeks after coronary stenting, however, they did not show any significant difference among the groups. CONCLUSION: Our study failed to demonstrate any beneficial effects of the short-term therapy with celecoxib and doxycycline or with celecoxib alone in the suppression of inflammatory biomarkers or in the inhibition of neointimal hyperplasia. Large scale randomized trials are necessary to define the role of anti-inflammatory therapy in the inhibition of neointimal hyperplasia.
Aged ; Angioplasty, Balloon, Coronary ; Anti-Bacterial Agents/therapeutic use ; Biological Markers/metabolism ; Coronary Artery Disease/immunology/metabolism/*therapy ; Cyclooxygenase 2 Inhibitors/therapeutic use ; Doxycycline/*therapeutic use ; Drug Therapy, Combination ; Female ; Follow-Up Studies ; Humans ; Male ; Metals ; Middle Aged ; Neointima/*drug therapy/*immunology/metabolism ; Pyrazoles/*therapeutic use ; Stents/*adverse effects ; Sulfonamides/*therapeutic use

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