臺灣博碩士論文加值系統

OxLDL已被指控為多個疾病的兇手，其中以心血管疾病中的粥狀動脈硬化(atherosclerosis)最盛。對於粥狀動脈硬化，人們的目光早已不在專注於血管上的故事，現在科學家更綜合地看心臟與血管環境。本研究論文旨在，藉由oxLDL刺激心肌H9c2細胞造成心肌肥大和凋亡，同時探討ZAKB對心臟的救援效果。過往人們認為是oxLDL先造成粥狀動脈硬化，接著心臟重塑，心肌肥大才產生，但現在的研究已經發現oxLDL可以直接對心臟造成傷害。同時，本研究也確實證明，oxLDL會誘導心肌肥大在10mg/dl oxLDL。這透漏人們過往對粥狀動脈硬化的治療是不完整的。接著從實驗室過往的發表中，我們發現ZAKB對減少心肌肥大造成的凋亡有著顯著的效果。因此，我們轉染ZAKB plasmid from Yang JJ(Chung Shan Medical University,台中,台灣)致心肌細胞中，實驗結果表明，ZAKB可以有效減少oxLDL造成的凋亡。並且我們還使用ZAK inhibitor D2825 from Ding K(J Med Chem. 2017 Jul 13,school of Pharmacy, Jinan University,China)加以證實ZAKB的重要性。實驗結果說明，當oxLDL刺激細胞凋亡時缺乏ZAKB的代償效果，將會使肥大的心肌細胞更快走向凋亡。這些結果顯示，ZAKB在幫助心臟承受來自oxLDL刺激時，ZAKB對於適應環境可能有著重要的地位。

OxLDL has been accused of being the culprit for multiple diseases, with the highest incidence of atherosclerosis in cardiovascular disease. For the atherosclerosis of the atherosclerosis, people's eyes have long been not focused on the blood vessels of the story, scientists now more comprehensive look at the heart and vascular environment. The aim of this study is to stimulate myocardial hypertrophy and apoptosis by oxLDL, and to explore the effect of ZAKB on heart rescue. In the past, people think that oxLDL first cause atherosclerosis, followed by cardiac remodeling, cardiac hypertrophy was produced, but now the study has found oxLDL can directly cause harm to the heart. At the same time, this study also confirmed that oxLDL induced cardiac hypertrophy at 10 mg / dl oxLDL. This is a breakthrough in the treatment of atherosclerosis in the past is not complete. Then from the past published in the laboratory, we found that ZAKB to reduce cardiac hypertrophy caused by apoptosis has a significant effect. Therefore, we transfected ZAKB plasmid from Yang JJ (Chung Shan Medical University, Taichung, Taiwan) to the cardiomyocytes, the experimental results show that, ZAKB can effectively reduce oxLDL caused by apoptosis. And we also confirmed the importance of ZAKB using ZAK inhibitor D2825 from Ding K (J Med Chem. 2017 Jul 13, school of Pharmacy, Jinan University, China). Experimental results show that if oxLDL stimulation of apoptosis when the lack of ZAKB compensatory effect, will make hypertrophic cardiomyocytes faster to apoptosis. These results show that ZAKB may play an important role in adapting to the environment when helping the heart to withstand the stimulation from oxLDL.

中文摘要 i
Abstract ii
Index iii
Table of figures v
Introduction 1
1-1 Cardiovascular diseases 1
1-2 Oxidized low density lipoprotein (OxLDL) 3
1-3 Cardiac Hypertrophy 7
1-4 Leucine-zipper and sterile-alpha motif kinase (ZAK) 9
Materials and methods 12
2-1. Cell culture 12
2-2. Cell subculture and cell counting 12
2-3. Cell frozen 12
2-4. Whole cell extraction 13
2-5. Lowry protein assay 13
2-6. MTT assay 13
2-7. Western blot analysis 14
2-8. Lipoprotein separation 14
2-9. TUNEL Assay 15
2-10. Statistical analysis 15
Result 16
3-1. OxLDL induced cell death in H9c2 cardiomyoblast cells. 16
3-2. Overexpression ZAKβ reduces apoptosis induced by oxLDL in H9c2 cardiomyoblast cells. 18
3-3. ZAK inhibitor D2825 and siZAKβ inhibits the rescue effect of ZAKβ in H9c2 cells treated with oxLDL. 20
Conclusion 22
References 23

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