ABSTRACT
Vitamin C (ascorbic acid) is an essential nutrient of most living tissues. We established a strain of Gulo-/- mice with known deficiency, in which vitamin C intake can be controlled by diet, like humans, and investigated the differentially expressed proteins following treatments with Helicobacter pylori and diethylnitrosamine (DENA) in the liver of Gulo-/- mice using a proteomic approach. Expression of p53, 14-3-3epsilon and 14-3-3delta in Gulo-/- mice liver tissue was analyzed by immunohistochemistry. 2-DE maps constructed from Gulo-/- mice liver and differentially expressed proteins in liver tissue were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF/MS). In Gulo-/- mice after H. Pylori infection, followed by treatment with DENA, no differences in p53, 14-3-3epsilon and 14-3-3delta were observed by immunohistochemistry. Proteome analyses using MALDI-TOF/MS resulted in successful identification of 12 proteins (nine proteins were up-regulated and three were down-regulated). Specifically, peroxiredoxin-6 and Alpha-1-antitrypsin 1-4 were up-regulated in liver after H. Pylori infection followed by treatment with DENA. These results indicated that oral supplementation with vitamin C led to rescue of Gulo-/- mice from vitamin deficiency, and protected the liver from H.pylori infection and/or DENA effect, and vitamin C also protected the liver against oxidative stress.
Subject(s)
Animals , Humans , Mice , Ascorbic Acid , Avitaminosis , Diet , Diethylnitrosamine , Helicobacter pylori , Helicobacter , Immunohistochemistry , Liver , Oxidative Stress , Proteins , ProteomeABSTRACT
OBJECTIVE: To evaluate and compare protein expression profiles of synovial fibroblasts using proteome analysis in swine knee injuries with joint instability, during early post-traumatic arthritis (PTA) development. METHOD: Experimental PTA was induced by transection of the anterior cruciate ligament (ACL) in swine left knee joints. After sacrifice at 8 weeks, cartilage and synovium obtained from both knee joints were prepared for histopathologic examination. Cultured synovial fibroblasts were processed for 2-dimensional electrophoresis and mass spectrometric analysis. Histopathologic examination showed overt arthritic changes that supported the development of early PTA. RESULTS: Proteome analyses led to the identification of more than 1,500 protein spots and of 11 differently expressed protein spots. Of those, six proteins were down-regulated (cytoskeletal beta actin, cofilin-1, destrin, Rho GDP dissociation inhibitor alpha, and unnamed protein product), and five proteins were up-regulated (alpha-B crystallin, smooth muscle protein 22-alpha, and cytoskeletal beta actin) in ACL-transected synovial fibroblasts. That is, proteins related to cellular organization and signal transduction are down-regulated, and those related to cell rescue, defence, and stress are up-regulated. CONCLUSION: These results may suggest that joint instability contributes to the development of PTA and is one of the major etiologic factors of PTA. In addition, this suggests that the proteome analysis of synovial fibroblasts is a useful approach in examining a joint after an injury and can be used to understand the pathogenesis of PTA.
Subject(s)
Actins , Anterior Cruciate Ligament , Arthritis , Cartilage , Crystallins , Destrin , Electrophoresis , Fibroblasts , Guanine Nucleotide Dissociation Inhibitors , Joint Instability , Joints , Knee Injuries , Knee Joint , Muscle, Smooth , Proteome , Signal Transduction , Swine , Synovial MembraneABSTRACT
PURPOSE: Breast cancer continues to be a major cause of death, despite the advances in the study of many prognostic factors. Although many prognostic factors have been studied, none reliably predict the response to treatment. This uncertainty in the prognostic factors could be overcome by defining the changes, occurring in patients at either the gene or protein level. Herein, attempts were made to examine the protein repertoire of patients using Proteomics. MATERIALS AND METHODS: Using conventional Proteomics, the high resolution 2-D electrophoresis followed by computational image analysis(Melanie program) and protein identification with mass spectrometry (MALDI-TOF), the serum of locally advanced breast cancer patients (stage III) was analyzed, and attempts were made to define the differences between recurred (or metastasis) patients ,and disease free patients of more than 4-years duration after surgery. RESULTS: In the 2-D electrophoresis of serum, about 1,000 spots were gained in each gel, with the up and down expressed protein spots compared to the normal control protein map. Six of seven patients had Cytokeratin 9 in their peripheral blood. In the serum of recurred patients (one of two), no Haptoglobin-related proteins were detected. All five un-recurred patients had normal or elevated levl of serum Haptoglobin-related proteins. CONCLUSIONS: The reduction of Haptoglobin-related proteins indicated the humoral immuno-depression in recurred patients. These findings may suggest the continuation of proper humoral immunity was important in the prevention of cancer recurrences or metastasis after surgery, especially in locally advanced breast cancer patients, which may suggests the value of immunotherapy in breast cancer patients to obtain good results.
Subject(s)
Humans , Breast Neoplasms , Breast , Cause of Death , Electrophoresis , Immunity, Humoral , Immunotherapy , Keratin-9 , Mass Spectrometry , Neoplasm Metastasis , Proteomics , Recurrence , UncertaintyABSTRACT
PURPOSE: Breast cancer continues to be a major cause of death, despite the advances in the study of many prognostic factors. Although many prognostic factors have been studied, none reliably predict the response to treatment. This uncertainty in the prognostic factors could be overcome by defining the changes, occurring in patients at either the gene or protein level. Herein, attempts were made to examine the protein repertoire of patients using Proteomics. MATERIALS AND METHODS: Using conventional Proteomics, the high resolution 2-D electrophoresis followed by computational image analysis(Melanie program) and protein identification with mass spectrometry (MALDI-TOF), the serum of locally advanced breast cancer patients (stage III) was analyzed, and attempts were made to define the differences between recurred (or metastasis) patients ,and disease free patients of more than 4-years duration after surgery. RESULTS: In the 2-D electrophoresis of serum, about 1,000 spots were gained in each gel, with the up and down expressed protein spots compared to the normal control protein map. Six of seven patients had Cytokeratin 9 in their peripheral blood. In the serum of recurred patients (one of two), no Haptoglobin-related proteins were detected. All five un-recurred patients had normal or elevated levl of serum Haptoglobin-related proteins. CONCLUSIONS: The reduction of Haptoglobin-related proteins indicated the humoral immuno-depression in recurred patients. These findings may suggest the continuation of proper humoral immunity was important in the prevention of cancer recurrences or metastasis after surgery, especially in locally advanced breast cancer patients, which may suggests the value of immunotherapy in breast cancer patients to obtain good results.