Mechanism of paeonol combined with paeoniflorin against myocardial ischemia injury:based on proteomics / 中国中药杂志

The study aims to investigate the effect of the compatibility of paeonol and paeoniflorin(hereinafter referred to as the compatibility) on the expression of myocardial proteins in rats with myocardial ischemia injury and explore the underlying mechanism of the compatibility against myocardial ischemia injury. First, the acute myocardial infarction rat model was established by ligation of the anterior descending branch of the left coronary artery. The model rats were given(ig) paeonol and paeoniflorin. Then protein samples were collected from rat cardiac tissue and quantified by tandem mass tags(TMT) to explore the differential proteins after drug intervention. The experimental results showed that differential proteins mainly involved phagocytosis engulfment, extracellular space, and antigen binding, as well as Kyoto encyclopedia of genes and genomes(KEGG) pathways of complement and coagulation cascades, syste-mic lupus erythematosus, and ribosome. In this study, the target proteins and related signaling pathways identified by differential proteomics may be the biological basis of the compatibility against myocardial ischemia injury in rats.

Method and application of proteomics in study of targets of traditional Chinese medicines / 中国中药杂志

The study on the targets of traditional Chinese medicine is an important part of researchers using modern scientific language to clarify the mechanism of traditional Chinese medicine. However, the research on the targets of Chinese medicine is full of challenges due to the complexity of active ingredients. As a branch of systems biology, proteomics focus on specific proteins in living organisms from a holistic perspective, which significantly improves the efficiency of targets discovery and has obvious advantages in the research of targets of Chinese medicine. Based on relevant literature and different methods used in targets of Chinese medicine, proteomics can be divided into chemical proteomics, differential proteomics and quantitative proteomics. The applications of the above three methods are illustrated in this paper as well, which will provide new methods and ideas for the study of the mechanism of Chinese medicine in the future.

Differential proteomics analysis on hydroquinone-induced cytotoxicity in TK6 cells / 中国职业医学

OBJECTIVE: To study the effect of hydroquinone on the protein expression on human lymphoblastoid cell TK6,and to explore the molecular mechanism of hydroquinone-induced cellular response. METHODS: The TK6 cells were treated with 20. 0 μmol/L of hydroquinone for 24. 0 hours. Total protein was extracted by protein lysis buffer and quantified. The proteins were separated by two-dimensional gel electroporthressis. After image analysis,the difference in electrophoresis was selected for enzymatic hydrolysis. The mass spectrometry identification was performed using matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry. The TK6 cells were treated with hydroquinone at a concentration of 0. 0,5. 0,10. 0 and 20. 0 μmol/L for 24. 0 hours,and total protein was extracted. The expression of heat shock protein 70( HSP70) and ubiquitin-binding enzyme 2( UBE2N) of which were identified by mass spectrometry were assayed by western blot. RESULTS: A total of 48 differential expression protein spots were detected after hydroquinone treatment,and the mass spectrometry identified 30 differentially expressed proteins with up-or down-regulation. These proteins were related to oxidative stress,mitochondrial energy metabolism,cytoskeleton,cell cycle,DNA damage repair,and so on. The relative expression levels of HSP70 and UBE2 N in TK6 cells of 5. 0,10. 0,20. 0 μmol/L hydroquinone group were higher than those of 0. 0 μmol/L hydroquinone group( P < 0. 05),which was consistent with the mass spectrometry results. CONCLUSION: Hydroquinone can induce cytotoxicity in TK6 cells through oxidative stress,which induces the change of mitochondrial energy metabolism and DNA damage repair.

Differential proteomic analysis of transgenic Bt rice and its non-transgenic parental rice / 国际药学研究杂志

Objective Under the methodology of differential proteomics and bioinformatics,the impact of the exogenous gene on the expression of rice in the proteome is discussed,aiming to explore into the study of genetically modified rice in the proteomics. Methods The total protein was extracted from genetically modified rice Huahui No.1(HH1) and non-transgenic rice Minghui 63 (MH63),the method of two-dimensional gel electrophoresis was applied to generate corresponding proteome two-dimensional poly?acrylamide gel(2D-PAGE)electrophoresis spectrum;then,the mass spectrometry and bioinformatics analysis were conducted after the selection of protein spots with significant differences. Results The comparing and matching of protein spots between transgenic Bt (cry1Ab/1Ac)rice and non-transgenic rice 2D-PAGE profiles identified 28 protein spots with significant differences. With non-trans?genic rice as a reference,transgenic Bt rice held 18 relatively high and 10 relatively low expressions;mass spectrometry and bioinfor?matics retrieval were made on the different protein spots. It was found that the differentiated protein was mainly involved in energy me?tabolism,protein synthesis,redox stress response and other biological processes. Conclusion Differences exist between transgenic Bt HH1 and its parental rice MH63 on the expression of proteome;however,there are neither anti-nutritional and allergenic protein, nor new or toxic proteins among these differentiated proteins.

Differential proteomic analysis of transgenic Bt rice and its non-transgenic parental rice / 国际药学研究杂志

Objective: Under the methodology of differential proteomics and bioinformatics, the impact of the exogenous gene on the expression of rice in the proteome is discussed, aiming to explore into the study of genetically modified rice in the proteomics. Methods: The total protein was extracted from genetically modified rice Huahui No.1(HH1) and non-transgenic rice Minghui 63(MH63), the method of two-dimensional gel electrophoresis was applied to generate corresponding proteome two-dimensional polyacrylamide gel(2D-PAGE) electrophoresis spectrum; then, the mass spectrometry and bioinformatics analysis were conducted after the selection of protein spots with significant differences. Results: The comparing and matching of protein spots between transgenic Bt(cry1Ab/1Ac) rice and non-transgenic rice 2D-PAGE profiles identified 28 protein spots with significant differences. With non-transgenic rice as a reference, transgenic Bt rice held 18 relatively high and 10 relatively low expressions; mass spectrometry and bioinformatics retrieval were made on the different protein spots. It was found that the differentiated protein was mainly involved in energy metabolism, protein synthesis, redox stress response and other biological processes. Conclusion: Differences exist between transgenic Bt HH1 and its parental rice MH63 on the expression of proteome; however, there are neither anti-nutritional and allergenic protein, nor new or toxic proteins among these differentiated proteins.

Approaches for targeted proteomics and its potential applications in neuroscience.

An extensive guide on practicable and significant quantitative proteomic approaches in neuroscience research is important not only because of the existing overwhelming limitations but also for gaining valuable understanding into brain function and deciphering proteomics from the workbench to the bedside. Early methodologies to understand the functioning of biological systems are now improving with high-throughput technologies, which allow analysis of various samples concurrently, or of thousand of analytes in a particular sample. Quantitative proteomic approaches include both gel-based and non-gel-based methods that can be further divided into different labelling approaches. This review will emphasize the role of existing technologies, their advantages and disadvantages, as well as their applications in neuroscience. This review will also discuss advanced approaches for targeted proteomics using isotope-coded affinity tag (ICAT) coupled with laser capture microdissection (LCM) followed by liquid chromatography tandem mass spectrometric (LC-MS/MS) analysis. This technology can further be extended to single cell proteomics in other areas of biological sciences and can be combined with other ‘omics’ approaches to reveal the mechanism of a cellular alterations. This approach may lead to further investigation in basic biology, disease analysis and surveillance, as well as drug discovery. Although numerous challenges still exist, we are confident that this approach will increase the understanding of pathological mechanisms involved in neuroendocrinology, neuropsychiatric and neurodegenerative disorders by delivering protein biomarker signatures for brain dysfunction.

Changes in leaf proteome profile of Arabidopsis thaliana in response to salicylic acid.

Salicylic acid (SA) has been implicated in determining the outcome of interactions between many plants and their pathogens. Global changes in response to this phytohormone have been observed at the transcript level, but little is known of how it induces changes in protein abundance. To this end we have investigated the effect of 1 mM SA on soluble proteins of Arabidopsis thaliana leaves by proteomic analysis. An initial study at transcript level has been performed on temporal landscape, which revealed that induction of most of the SA-responsive genes occurs within 3 to 6 h post treatment (HPT) and the expression peaked within 24 HPT. Two-dimensional gel electrophoresis (2-DE) coupled with MALDI-TOF MS/MS analysis has been used to identify differentially expressed proteins and 63 spots have been identified successfully. This comparative proteomic profiling of SA treated leaves versus control leaves demonstrated the changes of many defence related proteins like pathogenesis related protein 10a (PR10a), diseaseresistance- like protein, putative late blight-resistance protein, WRKY4, MYB4, etc. along with gross increase in the rate of energy production, while other general metabolism rate is slightly toned down, presumably signifying a transition from ‘normal mode’ to ‘defence mode’.