三黃瀉心湯對心臟缺血再灌流傷害動物模式之療效評估

葉竹來
高雄醫學大學
三黃瀉心湯（San-huang-xie-xin tang, SHXT）之三種組成植物大黃、黃蓮、黃芩而言，其組成成份已經證實具有抗癌、消炎及抗氧化的作用。其單味中藥成份Baicalein和Berberine許多研究文獻報導具有改善心臟血管的功能，可以用於治療高血壓、心律不整、心臟肥大和鬱血性心臟衰竭等疾病，然而對複方「三黃瀉心湯」並無相關文獻報導，因此本計畫之目的：擬以新標準化之三黃瀉心湯由靜脈投予，測試大鼠在左冠狀動脈阻塞45分鐘及再灌流120分鐘後心肌缺血再灌流傷害之改變，證實三黃瀉心湯具有防止心肌梗塞之心臟保護作用。實施方法：將大鼠隨機分成4大組，分別為控制組（Sham group）、溶媒組（Solvent-surgry group）、三黃瀉心湯靜脈注射組以及口服投予組，每組10隻大鼠。經由血行動力學變化、梗塞面積染色方法和血液creatine kinase (CK-MB)與 lactate dehydrogenase（LDH）之變化量，以測定三黃瀉心湯之作用程度。進一步探討三黃瀉心湯是否經由減低缺血再灌流引起心肌梗塞及細胞內程式凋亡。在每次實驗末期，將心臟進一步處理以利觀測細胞程式凋亡之進行及DNA漸層現象，運用in situ nick end-labeling (TUNEL) 的技術進行測定心臟細胞核內程式凋亡現象，測定光學顯微鏡下TUNEL標幟在心肌細胞核內顯微結構之變化；也將運用免疫墨點法測定Bcl-2、Bax、iNOS及caspases（-3及-9）的變化。在本計畫擬進一步以缺氧/再給氧引起大鼠H9c2心肌細胞凋亡模式，探討三黃瀉心湯可能具有抗細胞凋亡之心臟保護作用。由本計劃裡，我們期望經由上述之研究成果能確認三黃瀉心湯可以減低缺血再灌流引起的心臟損傷，並進一步提出證據說明，三黃瀉心湯可抑制心臟缺血再灌流時引發caspases活性增加及DNA片段產生現象，我們企圖期望三黃瀉心湯將來具有運用於心臟血管疾病臨床治療的潛在性。
關鍵字：三黃瀉心湯；心肌再灌流；細胞程式凋亡；心臟保護作用

Study the cardioprotection of San-Huang-Xie-Xin-Tang in ischemia-reperfusion injury rat model

Yeh, Jwu-Lai
Kaohsiung Medical University
San-Huang-Xie-Xin-Tang (SHXT) is a traditional Chinese medicinal formula containing Coptidis rhizoma, Scutellariae radix and Rheirhizoma. Recent studies suggest that the baicalein and berberine both exert beneficial effects on myocardial ischemia/reperfusion injury. Therefore, the purposes of this 1-year proposal are: (1) to accurate assessment of the safety and pharmacological effects of SHXT on post-ischemic cardiac function and dysrhythmias in ischemic/reperfusion myocardium, and (2) to clarify whether the cardioprotective effects include inhibition of apoptosis in cardiomyocytes and, if so, to identify the underlying mechanism(s). We will apply the SHXT against myocardial ischemia and reperfusion injury in anesthetized rats underwent 45 minutes of left coronary artery occlusion and 120 minutes of reperfusion. Methods: We will randomly separate the rats into 4 groups: sham, solvent, intravenous and oral SHXT groups. The incidence of reperfusion-induced hemodynamic changes (such as mean systemic arterial pressure and heart rate) and levels of infract area、creatine kinase (CK-MB) and lactate dehydrogenase (LDH) are measured. At the end of each experiment, hearts are processed for the evaluation of apoptosis and DNA laddering. The in situ nick end-labeling (TUNEL) technique is used to detect apoptotic cardiomyocyte nuclei. We will examine TUNEL at the light- and electron-microscopic level and analysis the nuclear ultrastructure of myocytes.
關鍵字：San-Huang-Xie-Xin-Tang；Myocardial reperfusion；Apoptosis；Cardioprotection