1 European Journal of Biotechnology and Bioscience Online ISSN: 2321-9122 www.biosciencejournals.com Volume 3; Issue 11; November 2015; Page No. 01-06 Fibroblast-specific protein-1(FSP-1) expression and metabolic alteration in aortic valve calcification 1 Randa S Hana, 2 Mohamed AK Salama Ayyad, 3 Heba A Yassa 1 Biochemistry Department 2 Cardiothorathic Surgery Department 3 Forensic and Clinical Toxicology Department. Assiut University, Egypt Abstract Objectives: Study the effect of hyperleptinaemia (systemically and locally) and hyperglyceamia on Fibroblast-specific protein-1 gene expression, atrial natriuretic peptide (ANP) level, and extent of calcification in stenotic aortic valves, the role of FSP1 as a marker of valve calcification, the role of aortic valve repair in ameliorating the metabolic alterations and study the sensitivity, specificity of ANP, leptin and NO as diagnostic and prognostic markers in valve diseases. Design and Methods: FSP-1(by qRT-PCR and western blotting), atrial natriuretic peptide, glucose, insulin and leptin (by ELISA), nitric oxide and calcium (colorimetrically) were studied in 20 aortic stenosis patients and 10 controls. Results: In valve tissues, high leptin was significantly associated with increased FSP1 and calcium deposition, 66% of patients had glucose intolerance with increase in the studied parameters and decreased to 36% after surgery. Conclusion: FSP-1 is a useful marker for valve calcification. Valve surgery, by decreasing leptin, ANP and NO had a beneficial role in improving insulin resistance, Elevated leptin and ANP levels, not insulin resistance, were associated with poor survival. Keywords: Fibroblast-specific protein-1, Atrial Natriuretic Peptide, Diseases Introduction Valve calcification has been most commonly described in the vessels of patients with diabetes. High glucose induced human aortic endothelial damage, endothelial-to-mesenchymal transition and contributed to calcification in diabetic rats via endothelial markers such as CD31 and fibroblast markers such as FSP-1, CD44 and CD10 [1, 2] . Fibroblast specific protein (FSP1) is a member of the S100 superfamily expressed by fibroblasts in fibrotic tissues mediating epithelial to mesenchymal transition that is up regulated in the diseased myocardium. Increased FSP-1 expression in smooth muscle cells was associated with their enhanced proliferation [3, 4] . The natriuretic peptides are endogenous cardiac hormones that include atrial natriuretic peptide (ANP) [5-7] , ANP is a powerful vasodilator, and a protein secreted from the heart muscle in response to atrial distention. Natriuretic peptide levels correlate with left ventricular (LV) function, are independent prognostic markers of cardiovascular diseases [8] , and are strong, independent predictors of sudden death in patients with cardiac failure [9] . Leptin, a hormone secreted by fat cells, is an important part of weight regulation. Leptin increases with obesity and decrease with weight loss. leptin level correlates with insulin level (both are high in hyperinsulinemia), in overweight individuals, physiological regulation of body weight by leptin seems to be disturbed, representing “leptin resistance.” This leptin resistance at the level of the pancreatic cell results in dysregulation of the adipoinsular axis and the development of hyperinsulinemia, obesity and diabetes [10] . In vitro studies demonstrated that leptin stimulates inducible NO synthase and generate increased amounts of NO that promote atherogenesis by inducing oxidative stress [11] .When NO is exposed to free radicals generated from oxidative stress, it undergo conversion to toxic molecules such as peroxynitrite that impair endothelial function [12] . Leptin at obese-range concentrations (but not at physiological one) impairs NO dependent vasorelaxation both in vitro and in vivo [13, 14] . Leptin exerts many potentially atherogenic effects such as hypertrophy and proliferation of vascular smooth muscle cells and coronary artery calcifications. Leptin enhances vascular calcification by inducing vascular smooth muscle and fibroblast proliferation (possibly through FSP1) and migration, enhance the expression of intracellular adhesive molecules [15] . Aim of the work • Study the influence of hyperleptinaemia (systemically and locally) and hyperglyceamia on FSP1 gene expression, ANP level, and extent of calcification in stenotic aortic valves. • The role of FSP1 as a marker of valve calcification and so the potential usefulness of FSP1 gene targeting in valve disease. • The role of aortic valve repair/replacement in ameliorating the metabolic alterations. • Study the sensitivity and specificity of ANP, leptin and NO as diagnostic and prognostic markers in valve diseases Patients and methods Twenty patients who had aortic valve stenosis assessed at Assiut university Hospital with a mean age of 70 years (between 66 and 73 years of age, 12 males and 8 females), overweight (between 70-100kgm) with a mean of 85.1±15.18kgm (mean BMI 28.3±4.027). The exclusion criteria