OBJECTIVETo investigate the expression of cyclooxygenase-2 (cox-2) in the testes and epididymides of adult male rats and its significance.

METHODSImmunohistochemical staining was used to detect the expression and localization of cox-2 in the testicle and epididymal tissues of 40 adult male SD rats.

RESULTSStrong cox-2 immunoreactivity was detected in the epididymides and testes of the rats. In the caput epididymides, cox-2 expressed mainly in the epithelial nuclei and partly in the cytoplasm. Cox-2 was also found positive in the testis nuclei and cytoplasm.

CONCLUSIONImmunohistochemical staining is a fairly sensitive method for detecting cox-2 expression in the testes and epididymides of adult male rats.

Animals ; Cyclooxygenase 2 ; biosynthesis ; Epididymis ; enzymology ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Sensitivity and Specificity ; Testis ; enzymology

OBJECTIVE: To investigate the expression of cyclooxygense-2 (COX-2) in eutopic and ectopic endometrium in ovarian endometriosis. METHODS: Thirty patients with ovrian endometriosis, 10 with ovarian chocolate cysts and 27 normal controls were enrolled it determine the expression of COX-2 immunohistochemically in eutopic or ectopic endometrium or healthy endometrial tissues. RESULTS: The immunoreactivities of COX-2 were found in epithelial cells and stromal cells in eutopic endometrium. The expression of COX-2 in the epithelial cells in the secretory phase was higher than that in the proliferative phase in the control group and ovarian endometriosis group (P <0. 05). But the expression of COX-2 in stromal cells in the control group and ovarian endometriosis group showed no cyclic changes throughout the menstrual cycle (P > 0. 05). The expression of COX-2 in eutopic and ectopic endometrium in the ovarian endometriosis group was higher than that in the control group (P <0. 05) , hut we did not find significant difference between the eutopic and ectopic endometrium in the ovarian endometriosis group (P > 0. 05). CONCLUSION The increased COX-2 expression in eutopic and ectopic endometrium in ovarian endometriosis may he related to its pathology.
Adult ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Endometriosis ; enzymology ; Endometrium ; enzymology ; Female ; Humans ; Immunohistochemistry ; Ovarian Diseases ; enzymology

Animals ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Female ; Liver Diseases, Alcoholic ; metabolism ; Male ; NF-kappa B ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley

In order to study the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in human laryngeal squamous cell carcinoma (LSCC) and its significance, the expression of VEGF mRNA and COX-2 mRNA in 62 cases of LSCC and 54 adjacent noncancerous laryngeal tissues and 9 normal human laryngeal mucous tissues was detected by using techniques of semi-quantitative RT-PCR. It was found that the expression level of VEGF and COX-2 mRNA was significantly increased in LSCC as compared with that in the normal human laryngeal mucous tissues (both P < 0.01), and the expression level of VEGF and COX-2 mRNA were significantly increased in stage Ill + IV tissues of LSCC as compared with the stage I + II tissues of LSCC (P < 0.01). There was a high positive correlation between VEGF and COX-2 expression in LSCC (r = 0.756, P < 0.01). These data raise the possibility that VEGF and COX-2 may play key roles in the growth, invasion and metastasis of LSCC.
Carcinoma, Squamous Cell/*metabolism ; Cyclooxygenase 2/*biosynthesis ; Cyclooxygenase 2/genetics ; Laryngeal Neoplasms/*metabolism ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Tumor Markers, Biological ; Vascular Endothelial Growth Factor A/*biosynthesis ; Vascular Endothelial Growth Factor A/genetics

OBJECTIVETo observe the changes in gene expression of cyclooxygenase (COX) during spontaneous recovery from stress ulcer in rats exposed to water immersion and restraint stress (WRS).

METHODSA rat model of stress ulcer was established by means of WRS, in which the changes in COX expression were detected with immunohistochemistry and reverse transcription (RT)-PCR.

RESULTSVery low levels of COX-2 expression were detected in the gastric mucosa of the control rats, and the expression increased significantly during the healing process of the stress ulcer (P<0.05). COX-1 expression in the gastric mucosa showed no significant difference between the control group and the stress ulcer groups during healing (P>0.05).

CONCLUSIONCOX-1 and COX-2 expressions in rat gastric mucosa during the recovery from stress ulcer participate in the recovery of the damaged mucosa possibly by mediating prostaglandin secretion.

Animals ; Cyclooxygenase 1 ; biosynthesis ; genetics ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Gastric Mucosa ; enzymology ; Male ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Remission, Spontaneous ; Stomach Ulcer ; enzymology ; Stress, Physiological ; complications

OBJECTIVETo investigate the relationship between the expression of COX-2 and liver cancer and construct a recombinant adenovirus encoding human COX-2 antisense RNA, and then to investigate its effects on liver cancer cell proliferation.

METHODSThe expression of COX-2 in 34 cases of hepatocellular carcinoma and in SMMC-7402 and SMMC-7721 cell lines was studied by using immunohistochemical techniques. The shuttle plasmid encoding anti-sense COX-2 was constructed by using cloning COX-2 cDNA fragment in the reverse direction into the pHCMVSPIA. Then the plasmid pJM17 and the shuttle plasmid were co-transferred into 293 cells with lipofectamine for homologous recombination to acquire recombinant adenovirus (Ad-AShcox-2), which was confirmed by PCR. Human hepatocellular carcinoma cell lines SMMC-7402 and SMMC-7721 were transduced in vitro. The cell apoptosis and cell cycle were analyzed by flow cytometry. The cell proliferation was determined by colony-forming efficiency.

RESULTSWe observed COX-2 expression in 82.4% of the hepatocellular carcinomas and SMMC-7402 cell line, but no COX-2 expression in the SMMC-7721 cell line. In addition, the recombinant adenovirus encoding anti-sense COX-2 fragment Ad-AShcox-2 was obtained with a titer of 1.06 x 10(12) PFU/ml. Ad-AShcox-2 reduced the expression of COX-2 and enhanced the percentage of cells into G1/G0 phase in the SMMC-7402 cell line. The difference of apoptotic index between the Ad-AShcox-2 group and the control group was statistically significant in SMMC-7402 but not in SMMC-7721. Similarly, colony-forming rates of SMMC-7402 and SMMC-7721 cell lines after Ad-AShcox-2 being transferred were (2.7+/-0.94)% and (33.6+/-4.24)%, respectively.

CONCLUSIONBy reducing the expression of COX-2 in hepatocellular carcinoma cells with the expression of COX-2, the cells could be inhibited.

Adenoviridae ; genetics ; Apoptosis ; physiology ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Humans ; Liver Neoplasms ; pathology ; Membrane Proteins ; biosynthesis ; genetics ; RNA, Antisense ; biosynthesis ; genetics

In order to investigate the expression of cyclooxygenase-2 (COX-2) in human lower segments of myometrium obtained from women in labor and those not in labor and identify the splicing variant of COX-2, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of COX-2. The primers were designed and synthesized according to the sequence of rat COX-2 splice variant which was discovered firstly by us. Then the splicing variant of COX-2 in human myometrium from woman in labor was identified, cloned into vector and sequenced. The results showed that the expression of COX-2 mRNA was lower in human myometrium obtained from women who were not in labor than that in labor women and a new band of COX-2 was obtained in myometrium from labor woman. The fragment included an unspliced intron, which pitched between exons 7 and 8. It was suggested that COX-2 gene was not only expressed highly in human myometrium from woman in labor, but also produced splicing variant by alternative splicing.
Amino Acid Sequence ; Base Sequence ; Cyclooxygenase 2/*biosynthesis ; Cyclooxygenase 2/genetics ; Labor Onset/*metabolism ; Molecular Sequence Data ; Myometrium/*enzymology ; Myometrium/metabolism ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Sequence Analysis

OBJECTIVETo observe in vitro the effect of Sinomenine, a pure alkaloid extracted from the chinese medical plant Sinomenium acutum on the activity of cyclooxygenase (COX-1 and COX-2) and the expression of COX-1 and COX-2 mRNA.

METHODMononuclear leukocytes were obtained from healthy adults. Isolated mononuclear leucocytes from human peripheral blood (PBMC) were incubated (1 x 10(6).mL-1) with or without sinomenine (or indomethacin), after incubated for 24 hours at 37 degrees C with 5% CO2; the media were assayed for the PGE2 by radioimmunoassay (RIA). LPS was used to stimulate the monocytes at a concentration of 5 micrograms.mL-1. And by RT-PCR, both COX-1 and COX-2 mRNAs were detected in Mononuclear leukocytes after incubation for different hours with drug (sinomenine or indomethacin) or not.

RESULTLPS (stimulated) induced the production of PGE2 in PBMC increasing with high expression of COX-2 mRNA; sinomenine reduced PGE2 production in LPS stimulated human monocytes more than in non-stimulated human monocytes. In comparative experiments, indomethacin, a non selective COX inhibitor, reduced the production of PGE2 equally in both states. Meanwhile, neither sinomenine(0.1-1 mmol.L-1) nor indomethacin(0.5-10 mumol.L-1) inhibited the expression of both COX-1 and COX-2 mRNAs by RT-PCR with beta-actin as reference.

CONCLUSIONIn contrast with indomethacin, Sinomenine shows a preferential inhibitory effect on COX-2 over COX-1, These results suggest that Sinomenine is a selective COX-2 inhibitor, which may be directly related to suppressing cyclooxygenase activity.

Adult ; Cyclooxygenase 1 ; Cyclooxygenase 2 ; Dinoprostone ; blood ; Humans ; Isoenzymes ; biosynthesis ; Leukocytes, Mononuclear ; enzymology ; Membrane Proteins ; Morphinans ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; metabolism ; RNA, Messenger ; biosynthesis ; Sinomenium ; chemistry

OBJECTIVETo investigate the changes of the mRNA expression and the intestinal mucosal cyclooxygenase (COXs) activity in scalded rats.

METHODSWistar rats inflicted with 30% TBSA III degree scalding were employed as the model. The changes of COX-1, COX-2 activities were determined by substrate fluorescence analysis and the mRNA expressions of COX-1 and COX-2 by RT-PCR.

RESULTSThe mRNA expressions and the activities of COX-2 in rat intestinal mucosa increased obviously after injury. But those of COX-1 exhibited lower range of change.

CONCLUSIONThe pathological mechanism of rat intestinal mucosa injury after scalding might be closely related to the COXs participation by different styles between the two enzymes.

Animals ; Burns ; enzymology ; genetics ; pathology ; Cyclooxygenase 1 ; Cyclooxygenase 2 ; Female ; Gene Expression ; Intestinal Mucosa ; enzymology ; Isoenzymes ; biosynthesis ; genetics ; metabolism ; Male ; Membrane Proteins ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Wistar

OBJECTIVETo investigate anticancer effect and molecular mechanism of N-[(Cyclohexyloxy)-4-nitrophenyl] methanesulfonamide on HepG2 cells in vitro.

METHODSHepG2 cells were treated with various concentrations (100, 200, 300, 400 micromol/L) of NS-398 [selective for cyclooxygenase 2 (COX-2) inhibition]. Cell growth was measured by MTT method, DNA fragmentation gel analysis was used to analyze the apoptosis cells, DNA ploidy and apoptotic cell percentage were examined by flow cytometry (FCM). PGE2 concentration was measured by radioimmunoassay method. The expressions of COX-2 were also examined by Western blot analysis.

RESULTSNS-398 inhibited HepG2 cell proliferation and induced apoptosis in a concentration-dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased and quiescent G1 phase was accumulated with NS-398 concentration increasing. The IC50 of 24 hours was 300 micromol/L. The release of PGE2 was significantly reduced in HepG2 cells with the values of NS-398 being (0.70 +/- 0.02), (0.48 +/- 0.02), (0.29 +/- 0.01) and (0.18 +/- 0.01) respectively, as compared with control group (0.03 +/- 0.01). NS-398 could inhibit the activity and expression of COX-2, with higher concentration, it can significantly down-regulate the expression of COX-2 (t = 3.736, 1.623, 1.810, 2.587, P < 0.01).

CONCLUSIONNS-398 might significantly inhibit the proliferation of HepG2 cells and induce apoptosis. The mechanisms were related with the accumulation of quiescent G1 phase and the inhibition of COX-2 activity.

Apoptosis ; drug effects ; Cell Line, Tumor ; metabolism ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; biosynthesis ; Humans ; Nitrobenzenes ; pharmacology ; Sulfonamides ; pharmacology