Animal ; Ascorbic Acid/metabolism ; Helicobacter Infections/*prevention & control ; Helicobacter pylori/*physiology ; Human ; Nitroso Compounds/metabolism ; Stomach Neoplasms/*microbiology ; Support, Non-U.S. Gov't

PURPOSE: Recent evidence shows that nitric oxide (NO) may exhibit both pro-cancer and anti-cancer activities. The present study aimed to determine the differentially expressed proteins in NO-treated NIH/3T3 fibroblasts in order to investigate whether NO induces proteins with pro-cancer or anti-cancer effects. MATERIALS AND METHODS: The cells were treated with 300 microM of an NO donor 3,3-bis-(aminoethyl)-1-hydroxy-2-oxo-1-triazene (NOC-18) for 12 h. The changed protein patterns, which were separated by two-dimensional electrophoresis using pH gradients of 4-7, were conclusively identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis of the peptide digests. RESULTS: Seventeen differentially expressed proteins were identified in NOC-18-treated cells. Nine proteins [vinculin protein, keratin 19, ubiquitous tropomodulin, F-actin capping protein (alpha1 subunit), tropomyosin 3, 26S proteasome-associated pad1 homolog, T-complex protein 1 (epsilon subunit) N(G)-dimethylarginine dimethylaminohydrolase, and heat shock protein 90] were increased and eight proteins (heat shock protein 70, glucosidase II, lamin B1, calreticulin, nucleophosmin 1, microtubule-associated protein retinitis pigmentosa/end binding family member 1, 150 kD oxygen-regulated protein precursor, and heat shock 70-related protein albino or pale green 2) were decreased by NOC-18 in the cells. Thirteen proteins are related to the suppression of cancer cell proliferation, invasion, and metastasis while two proteins (heat shock protein 90 and N(G)-dimethylarginine dimethylaminohydrolase) are related to carcinogenesis. The functions of 150 kD oxygen-regulated protein precursor and T-complex protein 1 (epsilon subunit) are unknown in relation to carcinogenesis. CONCLUSION: Most proteins differentially expressed by NOC-18 are involved in inhibiting cancer development.
Animals ; Electrophoresis, Gel, Two-Dimensional/*methods ; Fibroblasts/*metabolism/pathology ; HSP70 Heat-Shock Proteins ; Humans ; Mice ; NIH 3T3 Cells ; Neoplasms/*metabolism/pathology ; Nitric Oxide Donors ; Nitroso Compounds ; Proteins/analysis/*metabolism ; Proteomics/*methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Cerebral Cortex ; drug effects ; metabolism ; pathology ; Female ; Hypoxia-Ischemia, Brain ; drug therapy ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide Synthase ; metabolism ; Nitroso Compounds ; therapeutic use ; Rats ; Rats, Wistar

OBJECTIVETo observe the effect of Radix Ginseng and Radix Ophiopogonis extract (SMF) on protein S-nitrosylation in rats myocardial with ischemia/reperfusion injury (MI/RI).

METHODMyocardial ischemia/reperfusion in rats were produced by occlusion of the left anterior descending coronary artery. To study the cardioprotective effects of SMF on the acute MI/RI rats, the serum levels of creatine kinase (CK), lactate dehydrogenase (LDH), and nitric oxide (NO) were determined. The change of the expression of endothelial nitric oxide synthase (eNOS) was detected by Western blot. The content of related S-nitrosylation proteins in myocardial tissue was measured by Biotin-Switch method.

RESULTSMF significantly decreased the serum levels of CK and LDH as well as increased the serum levels of NO and the expression of eNOS in myocardial tissue. The contents of S-nitrosylation proteins were significantly increased from (4.42 +/- 0.60) micromol x g(-1) to (8.78 +/- 1.37) micromol x g(-1). The molecular weight of the majority S-nitrosylation proteins were in the range of 90 x 10(3)-117 x 10(3).

CONCLUSIONIncreased expression of eNOS and NO induced by SMF may activate S-nitrosylation of many proteins in rat hearts. The change of the activities or functions of those proteins by S-nitrosylation may be an important mechanism for myocardial protective effects of SMF.

Animals ; Blotting, Western ; Creatine Kinase ; blood ; Drugs, Chinese Herbal ; pharmacology ; L-Lactate Dehydrogenase ; blood ; Male ; Myocardial Ischemia ; blood ; drug therapy ; metabolism ; Myocardium ; metabolism ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type III ; metabolism ; Nitrosation ; drug effects ; Nitroso Compounds ; metabolism ; Panax ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Curcumin, an active ingredient of dietary spice used in curry, has been shown to exhibit anti-oxidant, anti-inflammatory and anti-proliferative properties. Using EB directed differentiation protocol of H-9 human embryonic stem (ES) cells; we evaluated the effect of curcumin (0-20 μmol/L) in enhancing such differentiation. Our results using real time PCR, western blotting and immunostaining demonstrated that curcumin significantly increased the gene expression and protein levels of cardiac specific transcription factor NKx2.5, cardiac troponin I, myosin heavy chain, and endothelial nitric oxide synthase during ES cell differentiation. Furthermore, an NO donor enhanced the curcumin-mediated induction of NKx2.5 and other cardiac specific proteins. Incubation of cells with curcumin led to a dose dependent increase in intracellular nitrite to the same extent as giving an authentic NO donor. Functional assay for second messenger(s) cyclic AMP (cAMP) and cyclic GMP (cGMP) revealed that continuous presence of curcumin in differentiated cells induced a decrease in the baseline levels of cAMP but it significantly elevated baseline contents of cGMP. Curcumin addition to a cell free assay significantly suppressed cAMP and cGMP degradation in the extracts while long term treatment of intact cells with curcumin increased the rates of cAMP and cGMP degradation suggesting that this might be due to direct suppression of some cyclic nucleotide-degrading enzyme (phosphodiesterase) by curcumin. These studies demonstrate that polyphenol curcumin may be involved in differentiation of ES cells partly due to manipulation of nitric oxide signaling.
Animals ; Antioxidants ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Curcumin ; pharmacology ; Cyclic GMP ; metabolism ; Embryoid Bodies ; drug effects ; metabolism ; physiology ; Enzyme Activators ; pharmacology ; Gene Expression ; drug effects ; Guanylate Cyclase ; genetics ; metabolism ; Homeobox Protein Nkx-2.5 ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Mice ; Myosin Heavy Chains ; genetics ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Donors ; pharmacology ; Nitric Oxide Synthase Type III ; genetics ; metabolism ; Nitroso Compounds ; pharmacology ; Pyrazoles ; pharmacology ; Pyridines ; pharmacology ; Second Messenger Systems ; Transcription Factors ; genetics ; metabolism ; Troponin ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; metabolism