線粒體乙醛脫氫酶2在心肌保護(hù)中的作用及可能機(jī)制探討
發(fā)布時(shí)間：2018-11-14 13:45
【摘要】：目的：研究ALDH2對(duì)離體大鼠心肌缺血/復(fù)灌損傷的作用，進(jìn)一步探討ALDH2心肌保護(hù)作用是否與體內(nèi)一氧化氮的生成有關(guān)，并分析線粒體通透性轉(zhuǎn)換孔是否參與其中。方法：實(shí)驗(yàn)大鼠隨機(jī)分為缺血/復(fù)灌組(Ischemia and Reperfusion, I/R)、酒精(ethanol, EtOH)干預(yù)組(I/R+EtOH)、氨基氰（cyanamide, CYA）干預(yù)組（I/R+CYA）及酒精+氨基氰干預(yù)組(I/R+EtOH+CYA)；硝基L-精氨酸甲基酯(NG-nitro-L-methyl arginine ester,L-NAME)干預(yù)組（I/R+L-NAME）及酒精+硝基L-精氨酸甲基酯干預(yù)組（I/R+EtOH+L-NAME）；酒精+蒼術(shù)苷(Atractyloside,Atr)干預(yù)組(I/R+EtOH+Atr)。采用離體大鼠心臟Langendorff灌流方法,結(jié)扎冠狀動(dòng)脈左前降支30min和復(fù)灌120min復(fù)制局部缺血/復(fù)灌損傷模型,測(cè)定左室心功能指標(biāo)；測(cè)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(cè)定心肌梗死面積； RT-PCR技術(shù)檢測(cè)心肌組織中ALDH2、Bcl-2、Bax的基因表達(dá)。結(jié)果：1.左室心功能指標(biāo)：I/R組缺血后左室發(fā)展壓（left ventricular developed pressure, LVDP）、左室內(nèi)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(fù)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(jìn)了復(fù)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(jié)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(fù)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(fù)灌5min時(shí)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(jìn)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(dá)顯著增高（P0.01），Bax表達(dá)顯著降低（P0.01）；CYA組和EtOH+CYA組ALDH2、Bcl-2表達(dá)顯著降低（P0.05~P0.01），Bax表達(dá)顯著增高（P0.05）；EtOH+Atr組ALDH2、Bcl-2表達(dá)顯著降低（P0.01），Bax差異無(wú)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(dá)顯著降低（P0.01），Bax表達(dá)顯著增高（P0.01）。結(jié)論：ALDH2活性增高起到保護(hù)心肌和抗凋亡的作用，ALDH2活性降低加重了心肌損傷，促進(jìn)心肌凋亡。ALDH2心肌保護(hù)機(jī)制可能與降低缺血/復(fù)灌心肌組織中的NO有關(guān)； ALDH2可抑制線粒體滲透性轉(zhuǎn)換孔道的開(kāi)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(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R363
本文編號(hào)：2331332
[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(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R363
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