【摘要】：目的：研究ALDH2對離體大鼠心肌缺血/復灌損傷的作用，進一步探討ALDH2心肌保護作用是否與體內一氧化氮的生成有關，并分析線粒體通透性轉換孔是否參與其中。方法：實驗大鼠隨機分為缺血/復灌組(Ischemia and Reperfusion, I/R)、酒精(ethanol, EtOH)干預組(I/R+EtOH)、氨基氰（cyanamide, CYA）干預組（I/R+CYA）及酒精+氨基氰干預組(I/R+EtOH+CYA)；硝基L-精氨酸甲基酯(NG-nitro-L-methyl arginine ester,L-NAME)干預組（I/R+L-NAME）及酒精+硝基L-精氨酸甲基酯干預組（I/R+EtOH+L-NAME）；酒精+蒼術苷(Atractyloside,Atr)干預組(I/R+EtOH+Atr)。采用離體大鼠心臟Langendorff灌流方法,結扎冠狀動脈左前降支30min和復灌120min復制局部缺血/復灌損傷模型,測定左室心功能指標；測量冠脈流出液中乳酸脫氫酶(lactatede hydrogenase，LDH)含量及心肌組織一氧化氮（nitric oxide, NO）含量； TTC雙染法測定心肌梗死面積； RT-PCR技術檢測心肌組織中ALDH2、Bcl-2、Bax的基因表達。結果：1.左室心功能指標：I/R組缺血后左室發(fā)展壓（left ventricular developed pressure, LVDP）、左室內壓最大上升速率/下降速率（±dp/dtmax）、左心室做功（rate pressure product, RPP）下降，心率（heart rate, HR）稍下降，左室舒張末壓（left ventricular end diastolic pressure,LVEDP）抬高，復灌末LVDP、±dp/dtmax、HR和RPP均下降明顯，LVEDP明顯抬高。與I/R組相比，EtOH組抑制了LVDP、HR、±dp/dtmax及RPP的下降并抑制了LVEDP的抬高（P0.05~P0.01）；CYA組促進了復灌期LVDP、±dp/dtmax、RPP的下降并抬高了LVEDP（P0.05~P0.01）；EtOH+CYA組、EtOH+L-NAME及EtOH+Atr組的結果與CYA組類似（P0.05~P0.01）。2.冠脈流出液中LDH含量：與I/R組相比，EtOH組、EtOH+L-NAME組明顯降低了復灌初期冠脈流出液中LDH的釋放（P0.01）；CYA組和EtOH+CYA組明顯增加了LDH的釋放(P0.01)。與EtOH組相比，EtOH+CYA組、EtOH+Atr組明顯增加了LDH的釋放(P0.01)；EtOH+L-NAME組在復灌5min時明顯增加了冠脈流出液中LDH的釋放(P0.01)。3.心肌梗死面積：與I/R組相比，，EtOH組降低了心肌梗死面積（P0.05）；CYA組、EtOH+CYA組及EtOH+Atr組明顯增加了心肌梗死面積(P0.01)。與EtOH組相比，EtOH+CYA組、EtOH+L-NAME組、EtOH+Atr組均明顯增加了心肌梗死面積(P0.01)。4.心肌組織中的NO量：與I/R組相比，EtOH組、L-NAME組及EtOH+L-NAME組均明顯降低了心肌組織中的NO含量(P0.01)。與EtOH組相比，EtOH+L-NAME組進一步降低了NO的釋放。5.RT-PCR：與I/R組相比，EtOH組心肌組織中ALDH2、Bcl-2表達顯著增高（P0.01），Bax表達顯著降低（P0.01）；CYA組和EtOH+CYA組ALDH2、Bcl-2表達顯著降低（P0.05~P0.01），Bax表達顯著增高（P0.05）；EtOH+Atr組ALDH2、Bcl-2表達顯著降低（P0.01），Bax差異無顯著性（P0.05）。與EtOH組相比，EtOH+CYA組、EtOH+L-NAME組、EtOH+Atr組ALDH2、Bcl-2表達顯著降低（P0.01），Bax表達顯著增高（P0.01）。結論：ALDH2活性增高起到保護心肌和抗凋亡的作用，ALDH2活性降低加重了心肌損傷，促進心肌凋亡。ALDH2心肌保護機制可能與降低缺血/復灌心肌組織中的NO有關； ALDH2可抑制線粒體滲透性轉換孔道的開放。
[Abstract]:Aim: to study the effects of ALDH2 on myocardial ischemia / reperfusion injury in isolated rats and to explore whether the myocardial protective effect of ALDH2 is related to the production of nitric oxide in vivo and whether the mitochondrial permeability transition pore is involved in it. Methods: experimental rats were randomly divided into ischemia / reperfusion group (Ischemia and Reperfusion, I / R) and alcohol (ethanol, EtOH) intervention group (I / R EtOH), cyanamide,). CYA (I / R CYA) and I / R EtOH CYA); (I / R EtOH CYA);) Nitro L-arginine methyl ester (NG-nitro-L-methyl arginine ester,L-NAME) intervention group (I / R L-NAME) and alcohol nitro-L-arginine methyl ester intervention group (I / R EtOH L-NAME); Alcohol Atractyloside (Atractyloside,Atr) intervention group (I / R EtOH Atr).) The left ventricular cardiac function was measured by ligating the left anterior descending coronary artery (30min) and reperfusion (120min) by Langendorff perfusion in isolated rat heart. The contents of lactate dehydrogenase (lactatede hydrogenase,LDH) and nitric oxide (nitric oxide, NO) in myocardial tissue were measured, the myocardial infarction area was measured by TTC double staining, and the gene expression of ALDH2,Bcl-2,Bax in myocardial tissue was detected by RT-PCR technique. Results: 1. Left ventricular function index: left ventricular development pressure (left ventricular developed pressure, LVDP), left ventricular pressure (rate pressure product, RPP) and heart rate (heart rate, HR) decreased in I / R group after ischemia. Left ventricular end-diastolic pressure (left ventricular end diastolic pressure,LVEDP) elevated, LVDP, 鹵dp/dtmax,HR and RPP decreased significantly and LVEDP increased significantly at the end of reperfusion. Compared with I / R group, EtOH group inhibited the decrease of LVDP,HR, 鹵dp/dtmax and RPP and the elevation of LVEDP (P0.05~P0.01); CYA group promoted the decrease of LVDP, 鹵dp/dtmax,RPP and LVEDP (P0.05~P0.01); The results of EtOH CYA group, EtOH L-NAME group and EtOH Atr group were similar to that of CYA group (P0.05~P0.01). 2. LDH content in coronary effluents: compared with I / R group, EtOH group and EtOH L-NAME group significantly decreased the release of LDH in coronary effluents (P0.01); CYA group and EtOH CYA group significantly increased LDH release (P0.01). Compared with EtOH group, EtOH Atr group in, EtOH CYA group significantly increased the release of LDH (P0.01); EtOH L-NAME group significantly increased LDH release in coronary effluents during 5min reperfusion (P0.01). Myocardial infarction area: compared with I / R group, EtOH group decreased myocardial infarction area (P0.05); CYA group, EtOH CYA group and EtOH Atr group significantly increased myocardial infarction area (P0.01). Compared with EtOH group, EtOH Atr group in EtOH L-NAME group increased myocardial infarction area significantly (P0.01). The amount of NO in myocardial tissue: compared with I / R group, EtOH group, L-NAME group and EtOH L-NAME group significantly decreased the content of NO in myocardial tissue (P0.01). Compared with EtOH group, EtOH L-NAME group further decreased the release of NO. 5. RT-PCR: compared with I / R group, the expression of ALDH2,Bcl-2 in myocardial tissue of EtOH group was significantly higher (P0.01), Bax expression was significantly lower (P0.01); ALDH2,Bcl-2 expression in CYA group and EtOH CYA group was significantly decreased (P0.05~P0.01), Bax expression was significantly increased (P0.05) ALDH2,Bcl-2 expression in); EtOH Atr group was significantly decreased (P0.01), Bax was not significantly different (P0.05). Compared with EtOH group, the expression of ALDH2,Bcl-2 in, EtOH Atr group of EtOH L-NAME group was significantly lower than that in, EtOH CYA group (P0.01), Bax expression was significantly higher (P0.01). Conclusion: the increase of ALDH2 activity can protect myocardium and inhibit apoptosis, and the decrease of ALDH2 activity can aggravate myocardial injury and promote myocardial apoptosis. The mechanism of myocardial protection of ALDH2 may be related to the reduction of NO in ischemic / reperfusion myocardium. ALDH2 inhibited the opening of mitochondrial permeability transition channels.
【學位授予單位】：蚌埠醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R363

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