1.Pressure change-mediated effects on vasoactive protein of endothelial cells in the flow field in vitro.
Jia HU ; Bengui ZHANG ; Jiang WU ; Eryong ZHANG ; Weilin XU ; Yingkang SHI ; Yingqiang GUO
Journal of Biomedical Engineering 2010;27(4):795-800
Lining the inner surface of the walls of blood vessels, Endothelial cells (ECs) go beyond providing selective membrane to maintain the natural structure and function of vessels; they also synthesize varieties of vasoactive proteins to modify the pressure shift in the local flow field and hence they adapt the physiological activities of vessels. In this experiment, ELISA and RT-PCR technologies were adopted. We set up five different pressure loaded ECs groups,one non-activated cultured ECs group and one single shear stress loaded ECs group. Such a design was intended to demonstrate the effects of pressure shift on the expression of vasoactive protein synthesized by ECs [Endothelin-1(ET-1), endothelial Nitric Oxide Synthase (eNOS), Cyclooxygenase-2(COX-2) and Vascular Endothelial Growth Factor(VEGF)]. Our aim was to elucidate the mechanism of the pressure shift mediated dysfunction in ECs and the related dose-effect relationship. Based on these data, we suggest that ECs could modify the expression of vasoactive protein for adapting to the pressure shift in the local flow field; while in the process of--40 cmH2O induced ECs' dysfunction, the vasoactive proteins eNOS, COX-2 and VEGF play an important role in protecting ECs.
Cells, Cultured
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Cyclooxygenase 2
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genetics
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metabolism
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Endothelial Cells
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metabolism
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physiology
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Endothelin-1
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genetics
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metabolism
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Humans
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Nitric Oxide Synthase Type III
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genetics
;
metabolism
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Pressure
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RNA, Messenger
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genetics
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metabolism
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Vascular Endothelial Growth Factor A
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genetics
;
metabolism
2.Changes of the mRNA expression and the intestinal mucosal cyclooxygenase activity in scalded rats.
Chinese Journal of Burns 2002;18(4):210-212
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
3.Effects of conjugated linoleic acid on the expression of critical enzymes of linoleic acid metabolism in tumor cell.
Jing-shu ZHANG ; Bing-qing CHEN ; Yan-hui GAO ; Yan-mei YANG ; Xuan-lin WANG ; Yang SONG ; Wen-guang SUN
Chinese Journal of Preventive Medicine 2005;39(3):195-198
OBJECTIVESTo study the effects of c9,t11-conjugated linoleic acid (c9,t11-CLA) on critical enzymes of linoleic acid metabolism in stomach granular cell (SGC-7901).
METHODSSGC-7901 was treated with c9,t11-CLA by 200, 100, 50 or 25 micromol/L for 24 hours. The effects of c9,t11-CLA on the cell proliferation was measured by monotetrazolium and the expression of Delta6-desaturase, Delta5-desaturase, COX-1, COX-2, 5-LOX mRNA were measured by reverse transcription polymerase chain reaction (RT-PCR).
RESULTSAt a concentration of 200, 100, 50, or 25 micromol/L, c9,t11-CLA suppressed the proliferation of SGC-7901 by 54.3%, 20.5%, 10.5% and 2.93%. The c9,t11-CLA might decrease the expression of COX-2 mRNA, and increase the expression of Delta6-desaturase and COX-1 in SGC-7901, but might not affect Delta5-desaturase and 5-LOX.
CONCLUSIONThe effects of c9,t11-CLA on the COX and Delta6-desaturase might play an important role in mediating the ability of c9,t11-CLA as to inhibiting the proliferation of tumor cells, and the anti-cancer activity by c9,t11-CLA might be associated with the linoleic acid metabolism.
Cell Line, Tumor ; Cyclooxygenase 1 ; genetics ; metabolism ; Cyclooxygenase 2 ; genetics ; metabolism ; Enzymes ; genetics ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Enzymologic ; drug effects ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Linoleic Acids ; metabolism ; Linoleic Acids, Conjugated ; pharmacology ; Lipid Metabolism ; drug effects ; Lipoxygenase ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction
4.Expression of cyclooxygenase-2, hMLH1 and hMSH2 proteins, and their relationship with microsatellite instability in gastric carcinoma.
Wei-qing SONG ; Cai-li HAN ; Yi CHEN ; Yi-hua ZHANG ; Jin-ying WEI ; Yu LIU
Chinese Journal of Oncology 2005;27(11):660-662
OBJECTIVETo investigate the expression of cyclooxygenase-2 (COX-2), human mut-l homologue 1 (hMLH1) and human mut-s homologue 2 (hMSH2) proteins in human paired gastric carcinoma (GC) and adjacent normal mucosa, and analyze their relationship with microsatellite instability (MSI).
METHODSThe protein expressions were examined by western blotting. Five MSI loci were assessed by PCR.
RESULTSIn 30 surgically excised GC tissues, the overexpression rate of COX-2, the low expression rate of hMLH1 and hMSH2 were 66.7%, 40% and 33.3%, respectively. Significant differences were found when compared with those of adjacent normal mucosa (P < 0.05). MSI was detected in 13 GC. The number of MSI-H (MSI-High, > or = 2 loci), MSI-L (MSI-Low, only one locus), and MSS (microsatellite stable) were 9, 4 and 17, respectively. The number of low expression rates of COX-2, hMLH1 and hMSH2 in MSI-H were 6, 8 and 5, respectively. There were significant differences compared to that of MSS (P < 0.05).
CONCLUSIONThe results suggest that microsatellite instability pathway is probably involved in the carcinogenesis of gastric carcinoma, which is frequently accompanied by low expression of hMLH1 and hMSH2, and may be also by low expression of COX-2.
Adaptor Proteins, Signal Transducing ; biosynthesis ; genetics ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Humans ; Microsatellite Repeats ; genetics ; MutL Protein Homolog 1 ; MutL Proteins ; Neoplasm Proteins ; biosynthesis ; genetics ; Nuclear Proteins ; biosynthesis ; genetics ; Stomach Neoplasms ; genetics ; metabolism
5.The expression and significance of HIF-1alpha and COX-2 in laryngeal squamous cell carcinoma.
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2007;21(18):820-824
OBJECTIVE:
To investigate the protein and mRNA expression of HIF-1alpha and COX-2 in laryngeal squamous carcinoma, and their correlation with clinicopathologic parameters.
METHOD:
The protein and mRNA expression of HIF-1alpha and COX-2 in fresh laryngeal squamous carcinoma tissues and adjacent normal tissues from 76 patients were examined by immunohistochemistry and reverse transcription polymerase chain reaction.
RESULT:
1) The protein positive expression rate of HIF-1alpha and COX-2 in tumor samples were significantly higher than that in normal tissues (P < 0.05); The protein positive expression rate of HIF-1alpha were higher in moderately and poorly differentiated tumors than in well differentiated tumors (P < 0.05); While the protein positive expression rate of COX-2 were not associated with T stage (P > 0.05); 2) The mRNA positive expression rate of HIF-1alpha and COX-2 were 67.11% and 75.00% respectively, their expression were significantly higher in tumors than that in normal tissues (P < 0.05). The mRNA expression of COX-2 were not detected in normal tissues. The mRNA positive expression rate of HIF-1alpha were associated with T stage, but were not associated with differentiated clinicopathologic stage. 3) The protein positive expression rate of HIF-1alpha were associated with that of COX-2.
CONCLUSION
HIF-1alpha and COX-2 could up regulate each other to promote the development and metastasis of laryngeal squamous carcinoma.
Adult
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Aged
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Carcinoma, Squamous Cell
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genetics
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metabolism
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pathology
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Cyclooxygenase 2
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genetics
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Female
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Humans
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Hypoxia-Inducible Factor 1, alpha Subunit
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genetics
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Laryngeal Mucosa
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pathology
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Laryngeal Neoplasms
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genetics
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pathology
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Male
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Middle Aged
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Neoplasm Staging
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Prognosis
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RNA, Messenger
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genetics
6.Tumor interstitial fluid and gastric cancer metastasis: an experimental study to verify the hypothesis of "tumor-phlegm microenvironment".
Da-zhi SUN ; Jian-peng JIAO ; Da-wei JU ; Min YE ; Xuan ZHANG ; Jing-yu XU ; Ye LU ; Jin HE ; Pin-kang WEI ; Ming-hui YANG
Chinese journal of integrative medicine 2012;18(5):350-358
OBJECTIVETo extract tumor interstitial fluid (TIF) from MKN-45 gastric cancer which is similar to "muddy phlegm" in Chinese medicine and observe influences of MKN-45 tumor interstitial fluid (MKN-45 TIF) intervention on metastasis of gastric cancer and on the expressions of vascular endothelial growth factor (VEGF), kinase insert domain containing receptor (KDR), epithelial-cadherin (E-cad), cyclooxygenase-2 (COX-2), intercellular adhesion molecule-1 (ICAM-1) and telomerase genes and proteins in primary tumor tissue.
METHODSAn MKN-45 tumor-bearing model was established in 50 nude mice. The modeled animals were equally randomized to 5 groups: the simple tumor-bearing group (model group), the normal saline (NS) via tail vein injection (i.v.) group (NS i.v. group), MKN-45 TIF i.v. group (TIF i.v. group), NS intraperitoneal injection (i.p.) group (NS i.p. group), and MKN-45 TIF i.p. group (TIF i.p. group). The TIF and NS intervention groups received injection (i.p. or i.v.) of MKN-45 TIF or NS twice a week, 0.2 mL at a time. After 8 weeks, the primary tumors were removed, weighed and HE stained to observe tumor metastasis. The primary tumor tissues were analyzed by immunohistochemistry and real-time quantitative PCR to detect expressions of VEGF, KDR, E-cad, COX-2, ICAM-1, and telomerase genes and proteins in different groups.
RESULTSThere were significant differences in tumor weight between TIF intervention groups and the model and NS intervention groups. Tumor metastasis was observed in all 5 groups, but the tumor metastasis rate in TIF intervention groups was significantly higher than those in the model and NS intervention groups. The gene and protein expressions of gastric cancer-related factors VEGF, KDR, COX-2, ICAM-1 and telomerase were unregulated while the gene and protein expressions of E-cad were downregulated in TIF intervention groups.
CONCLUSIONSTIF promotes tumor growth, invasion and metastasis of gastric cancer. These findings provide preliminary experimental clues for verifying the hypothesis of "tumor-phlegm microenvironment".
Animals ; Cadherins ; genetics ; metabolism ; Cell Line, Tumor ; Cyclooxygenase 2 ; genetics ; metabolism ; Extracellular Fluid ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Male ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Stomach Neoplasms ; metabolism ; secondary ; Telomerase ; genetics ; metabolism ; Tumor Microenvironment ; physiology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism
7.Cyclooxygenase-2 inhibitors modulate skin aging in a catalytic activity-independent manner.
Mi Eun LEE ; So Ra KIM ; Seungkoo LEE ; Yu Jin JUNG ; Sun Shim CHOI ; Woo Jin KIM ; Jeong A HAN
Experimental & Molecular Medicine 2012;44(9):536-544
It has been proposed that the pro-inflammatory catalytic activity of cyclooxygenase-2 (COX-2) plays a key role in the aging process. However, it remains unclear whether the COX-2 activity is a causal factor for aging and whether COX-2 inhibitors could prevent aging. We here examined the effect of COX-2 inhibitors on aging in the intrinsic skin aging model of hairless mice. We observed that among two selective COX-2 inhibitors and one non-selective COX inhibitor studied, only NS-398 inhibited skin aging, while celecoxib and aspirin accelerated skin aging. In addition, NS-398 reduced the expression of p53 and p16, whereas celecoxib and aspirin enhanced their expression. We also found that the aging-modulating effect of the inhibitors is closely associated with the expression of type I procollagen and caveolin-1. These results suggest that pro-inflammatory catalytic activity of COX-2 is not a causal factor for aging at least in skin and that COX-2 inhibitors might modulate skin aging by regulating the expression of type I procollagen and caveolin-1.
Animals
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Aspirin/administration & dosage
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Catalysis
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Caveolin 1/genetics/metabolism
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Collagen Type I/genetics/metabolism
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*Cyclooxygenase 2/metabolism/physiology
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Cyclooxygenase 2 Inhibitors/*administration & dosage
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Gene Expression Regulation
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Mice
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Nitrobenzenes/*administration & dosage
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Pyrazoles/administration & dosage
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Skin Aging/*drug effects/physiology
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Sulfonamides/*administration & dosage
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Tumor Suppressor Protein p53/genetics/metabolism
8.Study on the mechanism of preventing alcholic liver disease by selective cyclooxygenase 2 inhibitor.
Jun-ying ZHOU ; Dong-fang ZHOU ; Ying-hui LIU ; Wei WANG ; Feng WEI
Chinese Journal of Hepatology 2009;17(7):559-560
Animals
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Celecoxib
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Cyclooxygenase 1
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genetics
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metabolism
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Cyclooxygenase 2
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genetics
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metabolism
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Cyclooxygenase 2 Inhibitors
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pharmacology
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therapeutic use
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Cytokines
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metabolism
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Disease Models, Animal
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Ethanol
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adverse effects
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Fatty Liver, Alcoholic
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pathology
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prevention & control
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Immunohistochemistry
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Liver
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drug effects
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metabolism
;
pathology
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Liver Cirrhosis, Alcoholic
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pathology
;
prevention & control
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Liver Diseases, Alcoholic
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pathology
;
prevention & control
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Pyrazoles
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pharmacology
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therapeutic use
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RNA, Messenger
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genetics
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metabolism
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Rats
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Reverse Transcriptase Polymerase Chain Reaction
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Sulfonamides
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pharmacology
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therapeutic use
9.Anti-inflammatory mechanism of total glycosides of Acanthopanax Giraldii.
Fang YUAN ; Jie CHEN ; Guo-min XU ; Jia-jia ZHENG ; Qi-cai LONG
Chinese journal of integrative medicine 2009;15(3):210-215
OBJECTIVETo study the anti-inflammatory mechanisms of total glycosides of Acanthopanax Giraldii (TGA).
METHODSThe changes of prostaglandin E(2)(PGE(2)), tumor necrosis factor (TNF-alpha), nitric oxide (NO), and expressions of COX-1 mRNA and COX-2 mRNA in BALB/c mouse macrophages were observed by the radioimmunoassay, ELISA and nitric acid reduction and RT-PCR in the presence or absence of TGA.
RESULTS(1) TGA could significantly decrease the production of PGE(2)and NO in mouse peritoneal macrophages. The inhibitory rate to LPS-induced PGE(2)production was 87% (TGA 100 mg/L, P<0.05, vs. LPS) and 62% (TGA 20 mg/L, P<0.05, vs. LPS), respectively. The inhibitory rate of NO production in mouse peritoneal macrophages was 49% (TGA 100 mg/L, P<0.05, vs. LPS) and 21% (TGA 20 mg/L, P<0.05 vs. LPS), respectively. TGA could not inhibit LPS-induced TNF-alpha production in mouse peritoneal macrophages. (2) TGA also inhibited the expression of COX-1 and COX-2 mRNA in RAW264.7 cells. The inhibitory rate of TGA to COX-1 mRNA was 22% (TGA 100 mg/L, P<0.05, vs. blank). The inhibitory rate of TGA to COX-2 mRNA was 55% (TGA 20 mg/L, P<0.05, vs. LPS) and 100% (TGA 100 mg/L, P<0.01 vs. LPS), respectively.
CONCLUSIONThe anti-inflammatory mechanisms of TGA for inhibiting the production of NO and PGE(2)are through inhibiting COX-2 mRNA expression without TNF-alpha changes.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Cell Line ; Cyclooxygenase 1 ; genetics ; Cyclooxygenase 2 ; genetics ; Dinoprostone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Eleutherococcus ; Female ; Gene Expression Regulation, Enzymologic ; drug effects ; Glycosides ; pharmacology ; Lipopolysaccharides ; pharmacology ; Macrophages, Peritoneal ; cytology ; drug effects ; metabolism ; Mice ; Mice, Inbred BALB C ; Nitric Oxide ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism
10.Sympatric Distribution of Three Human Taenia Tapeworms Collected between 1935 and 2005 in Korea.
Hyeong Kyu JEON ; Kyu Heon KIM ; Jong Yil CHAI ; Hyun Jong YANG ; Han Jong RIM ; Keeseon S EOM
The Korean Journal of Parasitology 2008;46(4):235-241
Taeniasis has been known as one of the prevalent parasitic infections in Korea. Until recently, Taenia saginata had long been considered a dominant, and widely distributed species but epidemiological profiles of human Taenia species in Korea still remain unclear. In order to better understand distribution patterns of human Taenia tapeworms in Korea, partial nucleotide sequences of mitochondrial cox1 and ITS2 (internal transcribed spacer 2) were determined, along with morphological examinations, on 68 Taenia specimens obtained from university museum collections deposited since 1935. Genomic DNA was extracted from formalin-preserved specimens. Phylogenetic relationships among the genotypes (cox1 haplotype) detected in this study were inferred using the neighbor-joining method as a tree building method. Morphological and genetic analyses identified 3 specimens as T. solium, 51 specimens as T. asiatica, and 14 specimens as T. saginata. Our results indicate that all 3 Taenia tapeworms are sympatrically distributed in Korea with T. asiatica dominating over T. saginata and T. solium.
Animals
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Base Sequence
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Cestode Infections/epidemiology/parasitology
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Cyclooxygenase 1/genetics/metabolism
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DNA, Ribosomal Spacer/genetics
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Gene Expression Regulation
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Humans
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Korea/epidemiology
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Mitochondria/enzymology
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Molecular Sequence Data
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Phylogeny
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Taenia/*classification/*genetics
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Time Factors