.5 - Pyrroline Carboxylic Acid Reductase Expression In Rheumatic Mitral Tissue

Rheumatic fever/Rheumatic heart disease(RF/RHD) is a kind of systemic connective tissue disease and the cheif pathological changes of RHD are the progressive damages in valves.The valves are characterrized of a series of extracellular matrix (ECM) changes which are extensive proliferation and degeneration of collagen fibers, destruction and calcification of elastic fibers. Above changes result in stenosis and (or) regurgitation of the valves, thereby causing abnormal hemodynamic. Mitral and aortic valve are most commonly involved in RHD.P5CRs(pyrroline-5-carboxylate reductases) were currently found ubiquitously existed in eukaryotes. They are a kind of housekeeping proteins. In almost all cases, they catalyze the final step of proline synthesis, converting P5C (pyrroline-5-carboxylate) which is the intermediate metabolite of the proline, ornithine, arginine and glutamic acid into proline via an NAD(P)H-dependent reaction. The enzymatic cycle between P5C and proline is involved in the regulation of amino acid metabolism, intracellular redox potential, and apoptosis. Proline is an essential substance of connective tissue and the main component of collagen and elastin. The increased proline content plays an important role in rheumatic valvular extracellular matrix remodeling. This subject collected rheumatic heart disease, non-rheumatic mitral valve tissue and monocyte components. Experimental group consisted of 15 samples of rheumatic mitral valve and control group consisted of 7 nonrheumatic mitral valves from adults. To explore the pathological role and mechanisms of P5CR in patients' mitral valves with RHD, tissue samples were investigated by reverse transcription polymerasechain reaction (RT_PCR), western blot, immunohistochemistry and enzymatic detection.The main results are:(1) expression and purification the human pyrroline-5-carboxylate reductase(P5CR), (2) established the method of detecting the enzymatic activity of P5CR extraced from mitral tissue, (3) certified the expression of P5CR in rheumatic mitral tissue on the level of gene and molecular protein.