| BACKGROUND:In the post-genome era,the human genome project focus in the work of the functional gene research,with the different genetic screening technology continues to improve,the study of the disease mechanism are continued to deepen.Gene chip technology is the rise of a kind of molecular biology techniques in recent years.The technology has aroused widespread concern in the medical profession,application also more in the diagnosis and treatment of hypertension and its complications,has broad application prospects.CDNA microan'ayis a high-throughput,and rapid detection of gene expression means.In this study,using such new technologies to study hypertension blood stasis and non-blood stasis endothelial cells in patients to know more about the gene expression change and significance.Eukaryotic cytoskeleton fibers are formed by the three basic types of fiber and more than 100 binding protein.Apart from maintaining cell shape,cell separation fixed internal structures and participation in a certain cell,the eukaryotic cytoskeleton also has material transport,signal transduction,involved in cell movement,differentiation,proliferation and regulate gene expression effects.Since cytoskeletal functions are so diverse,then it changes will impact the cell behavior from the various.The cytoskeleton is quite sensitive to changes of the disease,a destructive role to microfilamentmake,thereby affecting cell growth and division,signal transduction,metabolic processes.The experiment conducted in hypertension blood stasis and non-blood stasis patients and healthy people cytoskeleton microfilament comparison.Discuss the changes in cytoskeleton between hypertension blood stasis or non-blood stasis and the normal control group.METHODS:experimental samples were divided into 3 groups:group hypertension blood stasis,hypertension non-blood stasis;normal control group.Application of gene chip technology we know the changes in gene expression of the endothelial cells between the two types of hypertension blood stasis and non-blood stasis patients,hypertension blood stasis and healthy people(the control group),then analysis the differences of gene expression. Total RNA synthesis Cy5/Cy3 labeled CDNA,a mixture of hybrid gene containing 8000 individual human high-density gene chips,after washing and scanning film we got signal and image,using computer analysis,by significantly standard,Including cell cycle proteins, apoptosis,cytoskeletal,sports proteins related genes and so on.Immunofluorescence staining with markers,they will be Morphology observed and photographed using confocal laser fluorescence microscope,cell actin expression quantitative analysis is also can be got.In this study F-actin Immunofluorescence staining with a ghost T cyclic peptide,observe and conduct fluorescence analysis by confocal laser fluorescence microscope.RESULTS:Hypertension blood stasis group demonstrated some kind of genes upward or downward.hypertension blood stasis existence of the dramatic changes in the cytoskeleton and F-actin content decreased.Cells array are changing direction,cell actin arranged in line with the direction of cells,cell actin fluorescence intensity decreased significantly compared to the normal control group;Conclusion:Hypertension serum with stasis caused significant difference to gene expression profiles of endothelial cells.to provide a new study direction for revealing blood stasis complex lesions mechanism,meanwhile,effective analysis of the gene expression using gene chip technology also helps understanding of the mechanism of drug treatment of blood stasis.Microfilament is the main component of cytoskeleton,play an important role in the life activity of the cells,the cytoskeleton microfilament observation quantitative and qualitative analysis of blood stasis is a good entry point for research in real terms,blood stasis modem research should be actively introduced the latest achievements of micro-structure and the gene science,provide the basis of molecular biology for the clinical treatment.
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