Effects of thrombopoietin on the protection against doxorubicin-induced cardiotoxicity.

To Man Yin.Thesis (M.Phil.)--Chinese University of Hong Kong, 2006.Includes bibliographical references (leaves 85-105).Abstracts in English and Chinese.Abstract (in English) --- p.i(in Chinese) --- p.ivAcknowledgements --- p.viPublications --- p.viiiTable of Contents --- p.ixList of Tables --- p.xiiList of Figures --- p.xiiiList of Abbreviations --- p.xivChapter CHAPTER 1: --- General Introduction --- p.1Chapter Section 1.1 --- Background and Clinical Application of Anthracylines --- p.1Chapter Section 1.2 --- DOX-induced Cardiotoxicity --- p.3Chapter 1.2.1 --- Types of Cardiotoxicity --- p.4Chapter 1.2.1.1 --- Acute Cardiotoxicity --- p.4Chapter 1.2.1.2 --- Chronic Cardiotoxicity --- p.5Chapter 1.2.2 --- Subcellular Effects of DOX --- p.6Chapter 1.2.2.1 --- Ultrastructural Lesions --- p.6Chapter 1.2.2.2 --- Effects on Mitochondrial Functions --- p.7Chapter 1.2.2.3 --- Effects on Sarcoplasmic reticulum (SR) Functions --- p.8Chapter Section 1.3 --- Mechanisms of DOX-induced Cardiotoxicity --- p.8Chapter 1.3.1 --- Formation of Free Radicals --- p.9Chapter 1.3.1.1 --- Generation of Free Radicals by DOX --- p.10Chapter 1.3.1.2 --- Cardiac damage by Free radicals --- p.12Chapter 1.3.2 --- Induction of Apoptosis --- p.14Chapter 1.3.2.1 --- Characteristics and Pathway of Apoptosis --- p.14Chapter 1.3.2.2 --- Mitochondria and Apoptosis --- p.15Chapter 1.3.2.3 --- Caspases and Apoptosis --- p.17Chapter 1.3.2.4 --- Apoptosis and DOX-induced Cardiotoxicity --- p.18Chapter Section 1.4 --- Strategies to Reduce DOX-induced Cardiotoxicity --- p.19Chapter 1.4.1 --- Dosage optimization and Schedule modification --- p.19Chapter 1.4.2 --- Anthracycline Analogues --- p.21Chapter 1.4.3 --- Cardioprotective Agents --- p.21Chapter Section 1.5 --- Thrombopoietin --- p.23Chapter CHAPTER 2: --- Hypotheses and Objectives --- p.30Chapter CHAPTER 3: --- Methodology --- p.33Chapter Section 3.1 --- Methods --- p.33Chapter 3.1.1 --- Culture of Rat H9C2 Myoblast Cell Line and Primary Neonatal Rat Cardiomyocytes --- p.33Chapter 3.1.1.1 --- Maintenance of Cell Line --- p.33Chapter 3.1.1.2 --- Culture of Primary Neonatal Rat Cardiomyocytes --- p.34Chapter 3.1.2 --- Effects of Thrombopoietin on Cell Viability of Rat H9C2 Myoblast Cell Line and Beating Rates of Primary Rat Cardiomyocytes --- p.35Chapter 3.1.2.1 --- Cell Viability assay --- p.35Chapter 3.1.2.2 --- Beating Rate of Primary Beating Cardiomyocytes --- p.36Chapter 3.1.3 --- Effects of Thrombopoietin on the Protection against DOX-induced Heart Injury In Vivo --- p.36Chapter 3.1.3.1 --- Animals --- p.36Chapter 3.1.3.2 --- Experimental Protocol --- p.37Chapter 3.1.3.3 --- Echocardiography --- p.38Chapter 3.1.3.4 --- Blood Cell Counts --- p.39Chapter 3.1.3.5 --- Histopathology --- p.39Chapter 3.1.4 --- Effects of Thrombopoietin on Apoptosis and Mitochondrial Integrity of Rat H9C2 Myoblast Cell Line and Apoptosis In Vivo --- p.40Chapter 3.1.4.1 --- Determination of Externalized Phosphatidylserine --- p.40Chapter 3.1.4.2 --- Determination of Active Caspase-3 Expression --- p.41Chapter 3.1.4.3 --- Assessment of Mitochondrial Integrity --- p.42Chapter 3.1.4.4 --- TUNEL assay --- p.43Chapter 3.1.5 --- Statistical Analysis --- p.44Chapter CHAPTER 4: --- Effects of Thrombopoietin on Cell Viability of Rat H9C2 Myoblast Cell Line and Beating Rates of Primary Neonatal Rat Cardiomyocytes --- p.46Chapter Section 4.1 --- Results --- p.46Chapter 4.1.1 --- Effects of TPO on DOX-induced Cell Death --- p.46Chapter 4.1.2 --- Effects of TPO on the Beating Rates of Primary Cardiomyocytes --- p.47Chapter Section 4.2 --- Discussion --- p.47Chapter CHAPTER 5： --- Effects of Thrombopoietin on the Protection Against DOX-induced Heart Injury In Vivo --- p.54Chapter Section 5.1 --- Results --- p.54Chapter 5.1.1 --- General Observations and Survival --- p.54Chapter 5.1.2 --- Blood Cell Counts --- p.55Chapter 5.1.3 --- Cardiac Functions by Echocardiography --- p.56Chapter 5.1.4 --- Gross Anatomic Changes and Pathology of the Myocardium --- p.57Chapter Section 5.2 --- Discussion --- p.58Chapter CHAPTER 6: --- Effects of Thrombopoietin on Apoptosis and Mitochondrial Integrity of H9C2 Cell Line and Apoptosis In Vico --- p.69Chapter Section 6.1 --- Results --- p.69Chapter 6.1.1 --- Determination of Externalized Phosphatidylserine --- p.69Chapter 6.1.2 --- Determination of Active Caspase-3 Activity --- p.70Chapter 6.1.3 --- Assessment of Mitochondrial Membrane Potential --- p.70Chapter 6.1.4 --- Determination of Apoptosis by TUNEL assay --- p.72Chapter Section 6.2 --- Discussion --- p.72Chapter CHAPTER 7： --- General Discussion and Conclusion --- p.83References --- p.8