PURPOSE: Cavernous nerve resection (CNR) in rats is a standard model of animal experiments on erectile dysfunction (ED) that occurs after radical prostatectomy (RP). Injured cavernous nerves after surgery can cause fibrosis and apoptosis that lead to penile structural changes that may be accompanied by alterations of protein expression. This study aimed to analyze the changes in protein after CNR in Wistar Kyoto rats. MATERIALS AND METHODS: Using 8-week-old male Wistar Kyoto rats, sham and CNR operation under a microscope were performed. Two and 8 weeks after surgery, we applied 2-DE and MALDI-TOF/TOF (AB 4700) to identify differently expressed penile proteins after CNR. 2-DE gels were stained with silver nitrate and were analyzed with PDQuest. After in-gel digestion, peptide mass spectra were obtained by MALDI-TOF/TOF mass spectrometry in the positive ion reflector mode. The obtained data were screened with a rat database from both the NCBI and the Swiss-Prot/TrFMBL home page. RESULTS: The proteins that were changed more than 1.5-fold compared with the sham group were annexin A4 and pyruvate kinase (PK). Annexin A4 was increased by 1.75-fold after 2 weeks, whereas PK was decreased by 4.16 after 8 weeks. These results were confirmed by immunohistochemistry. CONCLUSIONS: Annexin A4 in the CNR group was increased, which may be related to emiocytosis during apoptosis. The decrease in PK of the CNR group is assumed to be related to a decrease in efficacy during glycolysis. Further study will be needed to elucidate the molecular pathophysiology of ED after cavernous nerve injury.
Animal Experimentation ; Animals ; Annexin A4 ; Apoptosis ; Caves ; Digestion ; Erectile Dysfunction ; Fibrosis ; Gels ; Glycolysis ; Humans ; Immunohistochemistry ; Male ; Mass Spectrometry ; Prostatectomy ; Proteins ; Proteomics ; Pyruvate Kinase ; Rats ; Rats, Inbred WKY ; Salicylamides ; Silver Nitrate

Hepatic ischemia/reperfusion (I/R) injury is an inevitable consequence during liver surgery. I/R injury induces serious hepatic dysfunction and failure. In this study, we identified proteins that were differentially expressed between sham and I/R injured livers. Animals were subjected to hepatic ischemia for 1 hr and were sacrificed at 3hr after reperfusion. Serum ALT and AST levels were significantly increased in I/R-operated animals compared to those of sham-operated animals. Ischemic hepatic lobes of I/R-operated animals showed the hepatic lesion with unclear condensation and sinusoidal congestion. Proteins from hepatic tissue were separated using two dimensional gel electrophosresis. Protein spots with a greater than 2.5-fold change in intensity were identified by mass spectrometry. Among these proteins, glutaredoxin-3, peroxiredoxin-3, glyoxalase I, spermidine synthase, dynamin-1-like protein, annexin A4, eukaryotic initiation factor 3, eukaryotic initiation factor 4A-I, 26S proteasome, proteasome alpha 1, and proteasome beta 4 levels were significantly decreased in I/R-operated animals compared to those of sham-operated animals. These proteins are related to protein synthesis, cellular growth and stabilization, anti-oxidant action. Moreover, Western blot analysis confirmed that dynamin-1-like protein levels were decreased in I/R-operated animals. Our results suggest that hepatic I/R induces the hepatic cells damage by regulation of several proteins.
Animals ; Annexin A4 ; Blotting, Western ; Estrogens, Conjugated (USP) ; Eukaryotic Initiation Factor-3 ; Hepatocytes ; Ischemia ; Lactoylglutathione Lyase ; Liver ; Mass Spectrometry ; Mice ; Peptide Initiation Factors ; Proteasome Endopeptidase Complex ; Proteins ; Reperfusion ; Reperfusion Injury ; Salicylamides ; Spermidine Synthase

OBJECTIVE: To establish 2-dimensional electrophoresis (2-DE) graph of A549 and A549/DDP cell lines, to identify the differentially expressed proteins, and to screen multidrug resistance (MDR) related proteins in human lung adenocarcinoma. METHODS: The total proteins of A549 and A549/DDP cells were obtained, and were extracted and separated by 2-DE. PDQuest software was applied to analyze the 2-DE images, and the differential proteins of the 2 types of cells were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Western blot was used to determine the expression levels of the 4 proteins. RESULTS: We established 2-DE maps of total proteins from A549 and A549/DDP. A total of 40 differential protein spots in the 2 cell lines were found, and 23 differential expression proteins were identified by MALDI-TOF-MS. Western blot showed that heat shock protein beta-1, annexin A4, cofilin l, vimentin were differential expression proteins in A549 and A549/DDP, which was consistent with the results of the comparative proteomic analysis. CONCLUSION The 23 differential expression proteins in human lung adenocarcinoma are useful for studying the MDR mechanism of lung adenocarcinoma.
Adenocarcinoma ; genetics ; metabolism ; pathology ; Annexin A4 ; analysis ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Electrophoresis, Gel, Two-Dimensional ; HSP27 Heat-Shock Proteins ; analysis ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Proteins ; analysis ; classification ; Proteome ; analysis ; Proteomics ; methods

OBJECTIVE: Comparison of protein expression by two-dimensional gel electrophoresis (2-DE) in normal myometrium and uterine leiomyoma in Korean women. METHODS: Normal myometrium and uterine leiomyoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH4-8 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS- PAGE) and sliver stain. Scanned image analyzed using PDQuest 2-D softwareTM. Protein spot spectrum was identified by assisted laser desorption/ionization-time of fighting (MALDI-TOF) and the protein mass spectrums identification were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. RESULTS: In this study, we found 17 up-regulated proteins (phosphate carrier protein, 60 kDa heat shock protein, acidic calcium-independent, glutathione transferase omega, chloride intracellular channel 4, Ras-related protein Rab-11B, phosphatidylinositol transfer protein alpha isoform, type II keratin subunit protein, Cofilin 2 isoform 1, transgelin, ATP carrier protein, alpha-catenin homolog, parkinson disease 2, apo-cellular retinoic acid binding protein II, osteoglycin preproprotein, proteasome activator subunit 1 isoform, Unnamed protein) and 7 down-regulated proteins (Serum amyloid P component, annexin IV, alpha 1 actin precursor, hypoxanthine-guanine phosphoribosyltransferase, tumor necrosis factor receptor superfamily member EDAR precursor, peroxiredoxin 2, translation elongation factor EF-Tu precursor) between myometrium and leiomyoma. CONCLUSION: 2-DE offer total protein expression between normal myometrium and uterine leiomyoma, and searching of differently expressed protein for the diagnostic markers of leiomyoma.
Actins ; Adenosine Triphosphate ; alpha Catenin ; Animals ; Annexin A4 ; Carrier Proteins ; Cofilin 2 ; Databases, Protein ; Electrophoresis, Gel, Two-Dimensional* ; Electrophoresis, Polyacrylamide Gel ; Female ; Glutathione Transferase ; Heat-Shock Proteins ; Humans ; Hypoxanthine Phosphoribosyltransferase ; Isoelectric Focusing ; Keratins, Type II ; Leiomyoma* ; Mice ; Myometrium* ; Parkinsonian Disorders ; Peptide Elongation Factor Tu ; Peptide Elongation Factors ; Peroxiredoxins ; Phospholipid Transfer Proteins ; Proteasome Endopeptidase Complex ; Receptors, Tumor Necrosis Factor ; Running ; Serum Amyloid P-Component ; Sodium Dodecyl Sulfate ; Tretinoin