Thesis Final

  • View
    5

  • Download
    1

Embed Size (px)

Text of Thesis Final

  1. 1. 1 | P a g e Relationship of Nitric Oxide and Ascorbic Acid in Coronary Artery Disease Thesis submitted in partial fulfillment of the requirements for the degree of Masters of Science In Medical Biotechnology By: Priya Joshi Reg. No.: 131702016 M.Sc. Medical Biotechnology School of Life Sciences Manipal University Project Advisor: Dr Kamna Srivastava Assistant Professor B.R. Ambedkar Center for Biomedical Research University of Delhi New Delhi July 2015 Department of Biotechnology School of Life Sciences Manipal University, Manipal
  2. 2. 2 | P a g e
  3. 3. 3 | P a g e Acknowledgement I would sincerely and wholeheartedly like to acknowledge many people for their help and co-operation. Firstly I would like to thank our Director, Professor K. Satyamoorthy for giving me the opportunity to work on my Masters dissertation. I would also like to thank my guide, Dr. Kamna Srivastava for guiding me throughout my project, providing me with constant guidance, patience, support, valuable suggestions and help at every stage of my project. She gave me constant encouragement and ideas from the beginning of the project till the very end of it. I express my sincere gratitude and a warm thank you to Mr. Sudhir Chandra for his help throughout my project. I would like to thank my seniors who were very keen to extend a helping hand. Last but not the least I would like to thank my family and friends for all their support and encouragement. Ms. Priya Joshi
  4. 4. 4 | P a g e Serial No. Content Page No. 1. Introduction Coronary artery disease Nitric oxide Ascorbic acid 2 2. Literature review Prevalence of CVD in the world Prevalence of CVD in India Genetic evaluation for coronary artery disease Genetic susceptibility to myocardial infarction and coronary artery disease Association of MEF2A gene polymorphism with coronary artery disease Lack of MEF2A mutations in coronary artery disease Gene therapy to treat cardiovascular disease Genome wide association studies in myocardial infarction and coronary artery disease Nitric oxide Biochemistry of nitric oxide Chemical structure of nitric oxide Nitric oxide synthase Structure of eNOS Role of HSP in regulation of eNOS is cardiovascular diseases Vitamin C affects thrombosis/fibrinolysis system and reactive hyperemia in patients with type II diabetes and coronary artery disease 11 3. Methods 27 4. Results 32 5. Discussion 40 6. Conclusion 42 7. References 44 8. Synopsis 48 List of figures 1. Figure explaining physiology of atherosclerosis. 2. Figure showing diagnosis, symptoms, and causes of CVD. 3. Inflammation, calcification and scar development in atherosclerosis. 4. Classic risk factors in the formation and progression of atherosclerotic plaque, and possible pathogenic mechanisms of coronary artery disease risk genes. 5. Physiology of nitric oxide. 6. Conversion of L-arginine to nitric oxide. 7. Canonical structures for the ascorbic anion. 8. Global burden of cardiovascular disease. 9. Restriction fragment length polymorphism for rs325400. 10. Restriction fragment length polymorphism for rs34851361. 11. Nucleotide and amino acid sequences of the repeat region in MEF2A. 12. Pedigree analysis of families affected by CAD.
  5. 5. 5 | P a g e 13. A model of interactions of NO with erythrocytes and cell free hemoglobin in an arterial blood vessel. 14. eNOS protein structure. 15. Reaction between nitric oxide and sulphanilamide to form azo compound. 16. Graph showing concentrations of nitric oxide in control samples. 17. Graph showing concentrations of nitric oxide in case samples. 18. Graph showing mean of control and cases (nitric oxide). 19. Graph showing concentrations of ascorbic acid in control samples. 20. Graph showing concentrations of ascorbic acid in control samples. 21. Graph showing mean of control and cases (Ascorbic acid). List of tables 1. Gene therapy targets for coronary heart disease. 2. Summary of clinical characteristics 3. Table shows the values of all the samples of nitric oxide as observed after reaction with Griess reagent. (controls) 4. Table shows the values of all the samples of nitric oxide as observed after reaction with Griess reagent. (cases) 5. Table shows the values of all the samples of ascorbic acid as observed after reaction with dTCS reagent. (controls) 6. Table shows the values of all the samples of ascorbic acid as observed after reaction with dTCS reagent. (cases) 7. Statistical analysis
  6. 6. 6 | P a g e Introduction
  7. 7. 7 | P a g e Coronary artery disease The worldwide weight of cardiovascular diseases (CVD) is quickly expanding because of a sharp ascent in the occurrence and predominance of the same in creating nations. In the course of recent years, the CVD rates have multiplied in India. Expanded hereditary affinity to create CVD and expanding pervasiveness of cardiovascular danger variables are the reasons proposed for expanded seriousness and degree of coronary supply route maladies in Indians. Atherosclerotic coronary course malady (CAD) keeps on being an essential financial issue. A few phone segments, including endothelial, Inflammatory and smooth muscle cells of the blood vessel divider, take an interest in the era of an atherosclerotic plaque, the development of which is in charge of a dynamic narrowing of the lumen of the influenced coronary vessel and the following lessening in oxygen rich blood stream to downstream heart muscle, which might then be intensely or chronically harmed. Cardiovascular malady incorporates various diverse infections including coronary illness, stroke and fringe vascular ailment, hypertension. A normal 17.3 million people kicked the container from CVDs in 2008, identifying with 30% of single overall end. Of these passings, a normal 7.3 million were a result of coronary ailment and 6.2 million were a direct result of stroke. Low and center pay nations are disproportionally influenced: more than 80% of CVD passings happen in low and center pay nations and happen just as in men and ladies. By 2030, just about 25 million individuals will pass on from CVDs, principally from coronary illness and stroke. 7.5 million Deaths every year, or 13% of all passings can be ascribed to raised circulatory strain. This incorporates 51% of passings because of stroke and 45% because of coronary supply route ailment. It may happen through an acquired inclination. In India CVD was the biggest reason for passings in guys (20.3%) and in addition females (16.9%) and prompted around 2 million passings every year. (Price et al, 2001)
  8. 8. 8 | P a g e Figure 1: Figure explaining physiology of atherosclerosis. (Source: http://gosouthonline.co.za/atherosclerosis-or-hardening-of-the-arteries/) Real free hazard elements for CAD are known not recognition, high blood cholesterol levels, hypertension, smoking, diabetes and heftiness. Regardless of advances in danger element administration on an epidemiological level, numerous people keep on succumbing to CAD. Different blood markers connected with expanded danger for death and cardiovascular endpoints have been recognized, however right now not very many have been demonstrated to have an analytic effect or imperative clinical ramifications that would influence understanding administration. Troponin is the favored biomarker to analyze intense MI, though CKMBmass is the second marker of decision if troponin is not accessible. It is vital to recollect that troponin is a biomarker of cardiovascular rot and not exclusively a biomarker for dead tissue. A few clinical conditions can bring about a hoisted troponin level, markers that ascent sooner than troponin may be more prescient than single confirmation troponin esteem. In this way, there is an awesome requirement for inventive biomarkers that can survey hazard for CAD, evaluate action of the atherosclerotic process, and aide assessment of treatment. (Skalen et al, 2002)
  9. 9. 9 | P a g e A few late studies have proposed that flowing microRNAs could be helpful as biomarkers for different human disappointment and incessant vascular malady. MicroRNAs (miRNAs) are an as of late perceived class of short (19-25 nts), single stranded, non-coding RNAs that deal with a mixed bag of cell limits through the debasement and translational requirement of mRNAs that contain correlative groupings. More than 1000 human miRNAs have been recognized, and, in tissues, miRNAs manage the declaration of qualities included in discriminating cell procedures, including separation, development, multiplication, and apoptosis. (Cox et al, 1989) D'Alessandra and associates have shown that flowing muscle inferred miRNAs may be valuable as biomarkers of intense myocardial localized necrosis (AMI). They further demonstrated that muscle enhanced miRNAs, for example, miR-1, miR-133a, miR-133b and miR-499-5p are raised in plasma got from mice after coronary supply route ligation and in addition in people with AMI. In trial AMI models and in patients, Wang et al demonstrated that the muscle enhanced miR-1, miR-133a and miR-499 and additionally the cardiovascular particular miR-208 are hoisted in plasma. Moreover, miR-499, miR-1, miR-133a/b and miR-208 were indicated towards being expanded in little partners of people after AMI. (Ludmer et al, 1986) Real free hazard elements for CAD are known not recognition, high blood cholesterol levels, hypertension, smoking, diabetes and heftiness. Regardless of advances in danger element administration on an epidemiological level, numerous people keep on succumbing to CAD. Different blood markers connected with expanded danger for death and cardiovascular endpoints have been recognized, however right now not very many have been demonstrated to have an analytic effect or imperative clinical ramifications that would influence understanding administration. Troponin is the favored biomarker to analyze intense MI, though CKMBmass is the second marker of decision if tropo