本文選題：慢性間歇性低壓低氧 + anandamide��； 參考：《河北醫(yī)科大學(xué)》2011年碩士論文

【摘要】：目的:大量研究表明,慢性間歇性低壓低氧(Chronic intermittent hypobaric hypoxia, CIHH)具有明顯的心臟保護(hù)作用,可增強(qiáng)心臟對(duì)缺血、缺氧的耐受性,減輕缺血/再灌注對(duì)心肌的損傷,促進(jìn)缺血/再灌注后心臟舒縮功能的恢復(fù),以及抗心律失常。有研究報(bào)道,CIHH可改變機(jī)體對(duì)某些藥物的反應(yīng)性,從而間接發(fā)揮作用。Anandamide(AEA)是一種內(nèi)源性大麻素物質(zhì),具有抗心肌缺血、抗心律失常等心臟保護(hù)作用。但不知CIHH對(duì)AEA心臟保護(hù)作用有何影響。本研究旨在利用大鼠離體心臟缺血/再灌模型,觀察CIHH對(duì)AEA抗心肌缺血/再灌損傷保護(hù)作用的影響,并探討其機(jī)制。 方法:雄性成年Sprague-Dawlay(SD)大鼠隨機(jī)分為5組:間歇性低氧14天組(CIHH14)、間歇性低氧28天組(CIHH28)、anandamide組(AEA)、間歇性低氧14天加AEA組(CIHH14+AEA)和對(duì)照組(CON)。CIHH組和CIHH+AEA大鼠置于低壓氧艙,分別接受14天、28天模擬5000米海拔高度(PB=404 mmHg, PO2=84 mmHg)的低氧處理,每天6小時(shí)。AEA和CON組大鼠除了不接受低氧處理外,其它處理均與間歇性低氧組動(dòng)物相同。 大鼠麻醉固定,迅速開(kāi)胸,取出心臟,應(yīng)用Langendorff離體心臟灌流裝置制備心肌缺血/再灌注模型,即首先穩(wěn)定灌流30 min,停灌30 min(缺血),然后復(fù)灌60 min(再灌注)。將液體小囊插入左心室,室內(nèi)壓力通過(guò)壓力換能器輸入記錄系統(tǒng),分別描記離體大鼠心臟基礎(chǔ)狀態(tài)和缺血/再灌注狀態(tài)下不同時(shí)間的心臟功能的變化。心功能指標(biāo)包括:左室發(fā)展壓(left ventricular systolic pressure, LVDP)、左室舒張末壓(left ventricular end diastolic pressure, LVEDP)和左室壓力最大變化速率(maximal positive and negative velocity of left ventrichlar pressure,±LVdp/dtmax)。 對(duì)離體大鼠心臟,分別在缺血前給予不同的藥物干預(yù)。⑴對(duì)AEA和CIHH14+AEA組大鼠離體心臟,在缺血前給予10 min的AEA(2 nM)預(yù)處理;⑵對(duì)部分CIHH14+AEA大鼠離體心臟,在給AEA前分別給予10 min的AEA CB2受體阻斷劑AM630(10 nM)和CB1受體阻斷劑AM251(10 nM)預(yù)處理;⑶對(duì)部分CON大鼠離體心臟在缺血前分別給予10 min的AM630(10 nM)和AM251(10 nM)預(yù)處理。 實(shí)驗(yàn)過(guò)程中,留取各監(jiān)測(cè)時(shí)間點(diǎn)心臟灌流漏出液(冠脈流量),利用分光光度計(jì)按照乳酸脫氫酶試劑盒方法測(cè)定其中乳酸脫氫酶(lactate dehydrogenase, LDH)含量。實(shí)驗(yàn)結(jié)束后,取下心臟,置于液氮中冰凍保存。通過(guò)黃嘌呤氧化酶法方法,測(cè)定左心室心肌中超氧化物岐化酶(superoxide dismudase, SOD)活力,通過(guò)硫代巴比妥酸法計(jì)算出丙二醛(malondialdehyde, MDA)含量。 結(jié)果:⑴基礎(chǔ)狀態(tài)下,CIHH14、CIHH28、AEA、CIHH14+AEA和CON大鼠的LVDP分別為132.1±9.7、132.0±12.8、135.5±8.6、131.6±8.5和132.2±11.0;LVEDP分別為5.5±2.9、5.6±1.8、7.8±5.7、5.8±3.3和5.7±2.6;+LVdp/dtmax分別為3606.4±97.1、3616.8±109.2、3610.3±116.8、3586.6±90.4和3617.3±97.9 ; -LVdp/dtmax分別為2536.4±87.9、2612.0±46.4、2498.4±96.4、2480.4±63.0和2500.4±80.4,各組心功能參數(shù)間均無(wú)明顯差異(P0.05)。 ⑵缺血后、再灌注60 min時(shí),CIHH28和CIHH14+AEA大鼠的LVDP恢復(fù)分別為52.7±6.5%和53.1±8.5%,兩者間無(wú)顯著差異(P0.05),但明顯好于CIHH14大鼠的11.2±5.9%、AEA大鼠的8.9±5.3%和CON大鼠的10.4±4.0%(P0.01);LVEDP的恢復(fù)在CIHH28和CIHH14+AEA大鼠分別為48.3±12.3%和42.1±11.5%,兩者間無(wú)顯著差異(P0.05),但明顯好于CIHH14大鼠的14.5±10.2%、AEA大鼠的15.7±9.7%和CON大鼠的15.6±9.1%(P0.05);+LVdp/dtmax的恢復(fù)在CIHH28和CIHH14+AEA大鼠分別為38.1±2.2%和38.2±2.4%,兩者間無(wú)顯著差異(P0.05),但明顯好于CIHH14大鼠的8.6±1.5%、AEA大鼠的8.3%±1.1%和CON大鼠的8.5±1.5%(P0.05);-LVdp/dtmax的恢復(fù)在CIHH28和CIHH14+AEA大鼠分別為41.8±2.4%和40.5±1.3%,兩者間無(wú)顯著差異(P0.05),但明顯好于CIHH14大鼠的11.2±1.7%、AEA大鼠的11.5±2.1%和CON大鼠的10.7±1.3% (P0.05)。 ⑶缺血/再灌注后60 min時(shí),CIHH28和CIHH14+AEA大鼠冠脈流出液中的LDH分別為49.3±6.5和40.9±5.2,兩者間無(wú)明顯差異(P0.05),但明顯低于CIHH14、AEA和CON大鼠的93.7±6.4、110.2±8.0和101.6±8.0(P0.05);心肌組織中SOD在CIHH28和CIHH14+AEA大鼠分別為229.7±6.7和239.3±6.2,兩者間無(wú)明顯差異(P0.05),但明顯高于CIHH14、AEA和CON大鼠的160.2±6.7、170.7±7.3和181.9±4.4(P0.05);MDA在CIHH28和CIHH14+AEA大鼠分別為1.8±0.1和1.3±0.1,兩者間無(wú)明顯差異(P0.05),但明顯低于CIHH14、AEA和CON大鼠的2.1±0.1、2.0±0.1和2.2±0.1(P0.05)。 ⑷CB1受體阻斷劑AM251和CB2受體阻斷劑AM630對(duì)CON大鼠的心功能無(wú)影響。AM630可明顯抑制CIHH14+AEA大鼠心臟缺血后左心室功能的恢復(fù),使LVDP、LVEDP和±LVdp/dtmax恢復(fù)率明顯降低;同時(shí)冠脈流出液LDH升高,心肌組織SOD活性降低、MDA升高。而AM251無(wú)此作用。 結(jié)論:⑴28天CIHH處理具有明顯的心臟保護(hù)作用,可促進(jìn)大鼠缺血/再灌注后心功能的恢復(fù),降低冠脈流出液中的LDH含量,增加心肌組織SOD活性和降低心肌組織MDA含量。 ⑵14天CIHH處理和低濃度AEA對(duì)心臟均無(wú)明顯保護(hù)作用,但14天CIHH可增強(qiáng)AEA抗心肌缺血/再灌注損傷的心臟保護(hù)作用,促進(jìn)缺血/再灌注后心功能的恢復(fù),降低冠脈流出液中LDH含量,增加心肌組織SOD活性和降低心肌組織MDA含量。 ⑶CIHH對(duì)AEA的加強(qiáng)作用可被CB2受體阻斷劑AM630所阻斷。提示CIHH促進(jìn)AEA的心臟保護(hù)作用可能由CB2受體所介導(dǎo)。
[Abstract]:Objective: many studies show that chronic intermittent hypobaric hypoxia (Chronic intermittent hypobaric hypoxia, CIHH) has obvious protective effect on heart, can enhance heart to ischemia, hypoxia tolerance, reduce ischemia / reperfusion on myocardial ischemia / reperfusion injury and promote the recovery of cardiac function after injection, and anti arrhythmia. Studies have reported that CIHH can change the reactivity of some drugs, thereby indirectly play a role in.Anandamide (AEA) is a kind of endogenous cannabinoid substances with anti myocardial ischemia, cardiac protective effects of anti arrhythmia. But I do not know the effect of CIHH on the cardioprotective effects of AEA. The purpose of this study was to use rats from ischemia body / heart reperfusion injury model, observe the effect of CIHH on the protective effect of AEA against myocardial ischemia / reperfusion, and to explore its mechanism.
Methods: male Sprague-Dawlay (SD) rats were randomly divided into 5 groups: intermittent hypoxia group (CIHH14) 14 days, 28 days of intermittent hypoxia group (CIHH28), anandamide group (AEA), 14 days of intermittent hypoxia AEA group (CIHH14+AEA) and control group (CON) in group.CIHH and CIHH+AEA rats were placed in a hypobaric chamber respectively, for 14 days, 28 days of simulated 5000 meters altitude (PB=404 mmHg, PO2=84 mmHg) hypoxia treatment, 6 hours a day.AEA and CON rats in hypoxia exposure, other treatments with the same group of intermittent hypoxia animal.
The rats were anesthetized and fixed, quickly open the chest, remove the heart, using Langendorff isolated heart perfusion preparation of myocardial ischemia / reperfusion model, namely the first stable perfusion for 30 min, 30 min perfusion (ischemia), and 60 min of reperfusion (reperfusion). The liquid capsule was inserted into the left ventricle, the indoor pressure the pressure transducer input recording system, were recorded from the heart of the basic state and ischemia / reperfusion and heart function in different time under the condition of change. Including the indexes of cardiac function: left ventricular developed pressure (left ventricular systolic pressure, LVDP), left at the end of Shi Shuzhang (left ventricular end diastolic pressure pressure, and LVEDP) the maximum change rate of left ventricular pressure (maximal positive and negative velocity of left ventrichlar pressure + LVdp/dtmax).
On the isolated rat heart, were treated with different drug intervention before ischemia. The heart of AEA and CIHH14+AEA rats, given 10 min before ischemia AEA (2 nM) of the pretreatment; the heart part of CIHH14+AEA rats, to AEA were given 10 min AEA CB2 receptor antagonist AM630 (10 nM) and CB1 receptor antagonist AM251 (10 nM) on the part of the pretreatment; CON isolated rat heart before ischemia were treated with 10 min AM630 (10 nM) and AM251 (10 nM) pretreatment.
During the experiment, take the monitoring time heart perfusion transudate (coronary flow), using the spectrophotometer to measure the lactate dehydrogenase method (lactate dehydrogenase, according to Lactate Dehydrogenase Kit LDH) content. After the end of the experiment, take heart, kept frozen in liquid nitrogen. By the method of xanthine oxidase method, determination of left ventricular myocardium superoxide dismutase (superoxide dismudase, SOD) activity, calculated by the thiobarbituric acid method malondialdehyde (malondialdehyde, MDA) content.
Results: the basic state, CIHH14, CIHH28, AEA, CIHH14+AEA and CON rats LVDP were 132.1 + 9.7132.0 + 12.8135.5 + 8.6131.6 + 8.5 and 132.2 + 11; LVEDP = 5.5 + 2.9,5.6 + 1.8,7.8 + 5.7,5.8 + 3.3 and 5.7 + 2.6; +LVdp/dtmax = 3606.4 + 97.13616.8 + 109.23610.3 + 116.83586.6 + 90.4 and 3617.3 + 97.9; -LVdp/dtmax = 2536.4 + 87.92612.0 + 46.42498.4 + 96.42480.4 + 63 and 2500.4 + 80.4, there were no significant differences between the parameters of heart function (P0.05).
鈶電己琛，

本文編號(hào)：1760605