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研究生: 董威琳
論文名稱: 桂枝加味生脈飲預處理減少大鼠心臟缺血再灌流傷害
Gui-zhi Jiawei Sheng-mai-yin Pretreatment Reduces Cardiac Ischemia/Reperfusion Injury in the Rat
指導教授: 鄭劍廷
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2017
畢業學年度: 105
語文別: 英文
論文頁數: 52
中文關鍵詞: 桂枝生脈飲缺血再灌流傷害缺血性心臟病心肌梗死
英文關鍵詞: Gui-zhi, Sheng-mai-yin,, ischemia and reperfusion injury, ischemic heart disease, myocardial infarction
DOI URL: https://doi.org/10.6345/NTNU202203318
論文種類: 學術論文
相關次數: 點閱:132下載:11
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  • 心臟手術和心臟梗塞後可能導致局部缺血再灌流(I / R)損傷誘發心力衰竭,心臟病變甚至死亡。因此,研究預防性藥物或方法以降低I/R傷害程度和死亡率在心臟疾病研究中十分重要。生脈飲(SMY)是一種含有人參、麥冬和五味子的中國傳統複方,已被用於治療心血管疾病超過700年,但是其保護的機轉尚未被研究透徹。桂枝(GZ)也具有心臟保護的效果,我們期望加味桂枝能提高生脈飲的心臟保護效果。
    在這項研究中,我們管餵生脈飲或桂枝加味生脈飲7天作為預處理之後再結紮和鬆綁左升冠狀動脈,來探討生脈飲在雄性Wistar大鼠心臟I / R損傷的保護作用。實驗分為六組:對照組、SMY控制組、GZ+SMY控制組、I/R 控制組、SMY I/R組和GZ+SMY I/R組。在整個實驗過程中,心電圖(ECG)和血壓均連續地即時測量。左心室舒張末期壓(LVEDP)也被測量以評估心臟收縮力和功能。心肌梗死面積則是以MOOR表面血液影像和伊文思藍/ TTC雙染色呈現。而H&E染色則用來研究組織病理學之改變。結果顯示,I/R傷害會造成心電圖中ST段上升,心臟的LVEDP、梗死面積、紅血球累積和白血球浸潤增加。經過生脈飲預處理後皆有改善情況。但桂枝加味生脈飲預處理並不比生脈飲有更好效果。依據以上結果可知,桂枝加味生脈飲和生脈飲表現相似之保護能力可降低大鼠心肌缺血再灌流的損傷。

    Cardiac operations and heart infarctions may lead to ischemia and reperfusion (I/R) injuries that induce cardiac failure, morbidity, and mortality. Therefore, the use of preventive drugs or strategy to reduce high severity and high mortality is important in the cardiac disease research. Sheng-mai-yin (SMY) is one traditional Chinese formulation containing Radix Ginseng, Radix Ophiopogonis and Fructus Schisandrae that has been used to treat cardiovascular diseases for over 700 years. However, the protective mechanism is not fully understood. Gui-zhi (GZ) was also shown its cardioprotective ability. We aimed to test whether the SMY could attenuate I/R injury and whether the GZ remixed SMY (GZ+SMY) could have better effect than the only SMY.
    In this study, we used tube feeding SMY, GZ, or GZ+SMY for 7 days as a pretreatment to explore its protective effect in the male Wistar rats with cardiac I/R injury by left ascending coronary artery ligation and release. There were six groups in our experiment: sham control, SMY control, GZ+SMY control, I/R SMY I/R, and GZ+SMY I/R group. The electrocardiograph (ECG) and arterial blood pressure were continuously and real-time measured throughout the experiment. The left ventricular end-diastolic pressure (LVEDP) was also measured to evaluate the cardiac contractility and function. The myocardial infarct size was indicated by MOOR blood flow image and Evans blue/TTC staining. The H&E staining was used to investigate the histopathologic changes. I/R injury increased ST segment elevation, LVEDP level, infarct size, erythrocyte accumulation and leukocyte infiltration in the heart. Seven days of Sheng-mai-yin pretreatment significantly reduced the parameters of the LEVDP, infarct size, erythrocyte accumulation and leukocyte infiltration in I/R hearts compared to I/R group. However, GZ remixed SMY did not exert much better effect than SMY alone in our study. According to above data, we conclude that GZ+SMY displayed a similar effect as SMY to protect rat heart against I/R injuries.

    目錄 I ABSTRACT 3 II 中文摘要 5 III ABBREVIATIONS 7 IV INTRODUCTION 9 V MATERIALS AND METHODS 13 1. GROUPING 13 2. DRUGS PREPARATION 14 3. THE MODEL OF ISCHEMIA AND REPERFUSION IN RATS 14 4. LASER DOPPLER IMAGING 15 5. MEASUREMENT OF LEFT VENTRICULAR PRESSURE 16 6. DETERMINATION OF ANTI-OXIDATION ABILITY 16 7. TTC STAINING 17 8. HISTOPATHOLOGY 17 9. IMMUNOHISTOCHEMISTRY 17 10. TERMINAL DEOXYNUCLEOTIDE TRANSFERASE DUTP NICK END LABELING STAIN1 18 11. WESTERN BLOT 20 12. STATISTICAL ANALYSES 21 VI RESULTS 22 1. THE ANTIOXIDANT CAPACITY OF SHENG-MAI-YIN AND GUI-ZHI 22 2. ELECTROCARDIOGRAMS(ECG)RECORDING 22 3. LEFT VENTRICLE PRESSURE 22 4. MYOCARDIAL INFARCT SIZE 22 5. HISTOPATHOLOGY 23 6. IMMUNOHISTOCHEMISTRY 23 7. TERMINAL DEOXYNUCLEOTIDE TRANSFERASE DUTP NICK END LABELING STAIN 24 8. WESTERN BLOT 24 VII DISCUSSION 26 1. THE ANTIOXIDATIVE ABILITY OF SHENG-MAI-YIN AND GUI-ZHI 26 2. THE IMPROVEMENT OF HEART CONTRACTILE FUNCTION 26 3. THE INFARCT SIZE WAS DECREASED 27 4. HISTOPATHOLOGY 27 5. AUTOPHAGY WAS DECREASED BY SMY 27 6. APOPTOSIS WAS DECREASED BY SMY 28 7. THE RELATIONSHIP BETWEEN ENOS AND THE SMY 28 VIII CONCLUSION 30 IX FIGURES 31 FIGURE. 1 CHEMILUMINESCENCE ANALYZING 31 FIGURE. 2 ELECTROCARDIOGRAPH OF I/R GROUP 33 FIGURE. 3 THE LEFT VENTRICLE PRESSURE MEASUREMENT 34 FIGURE. 4 THE EVANS BLUE/TTC STAINING 35 FIGURE. 5 LASER DOPPLER IMAGING 36 FIGURE. 6 H&E STAINING 39 FIGURE. 7 IMMUNOHISTOCHEMISTRY OF 4-HNE 40 FIGURE. 8 IMMUNOHISTOCHEMISTRY OF BECLIN-1 41 FIGURE. 9 IMMUNOHISTOCHEMISTRY OF LC3 42 FIGURE. 10 TERMINAL DEOXYNUCLEOTIDE TRANSFERASE DUTP NICK END LABELING STAIN 43 FIGURE. 11 WESTERN BLOT OF BECLIN-1 45 FIGURE. 12 WESTERN BLOT OF C-PARP 46 FIGURE. 13 WESTERN BLOT OF ENOS 47 X REFERENCES 48

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