中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2015年
5期
632-636
,共5页
杜文娟%王海英%李小娟%陈伟%徐鹏%喻田
杜文娟%王海英%李小娟%陳偉%徐鵬%喻田
두문연%왕해영%리소연%진위%서붕%유전
异氟醚%脂肪乳剂,静脉注射用%缺血后处理%心肌再灌注损伤%NF-E2相关因子2%反应元件
異氟醚%脂肪乳劑,靜脈註射用%缺血後處理%心肌再灌註損傷%NF-E2相關因子2%反應元件
이불미%지방유제,정맥주사용%결혈후처리%심기재관주손상%NF-E2상관인자2%반응원건
Isoflurane%Fat emulsions,intravenous%Ischemic postconditioning%Myocardial reperfusion injury%Nuclear factor-E2 related factor 2%Response elements
目的：评价乳化异氟醚后处理对大鼠缺血再灌注时核转录因子 NF?E2相关因子2（ Nrf2）?抗氧化反应元件（ ARE）信号通路的影响。方法健康雄性SD大鼠，体重250～300 g，4～5月龄，采用Langendorff灌注装置建立大鼠离体心脏灌注模型。取模型制备成功的心脏32个，采用随机数字表法分为4组（ n＝8）：对照组（ C组）、缺血再灌注组（ I∕R组）、乳化异氟醚后处理组（ EIP组）和脂肪乳组（ F组）。平衡灌注20 min后，C组继续灌注100 min；I∕R组32℃下缺血40 min，恢复灌注60 min；EIP组32℃下缺血40 min，于再灌注前即刻灌注含乳化异氟醚1．68 mmol∕L的K?H液2 min，继续灌注37℃含氧的K?H液58 min；F组32℃下缺血40 min，于再灌注前即刻灌注含脂肪乳712 mg∕L 的K?H液2 min，继续灌注37℃含氧K?H液58 min。分别于平衡灌注末及再灌注末记录HR、左室发展压（LVDP）、左室舒张末压（LVEDP）和左心室压力最大上升速度（＋dp∕dtmax）。于再灌注末，取左心室心肌组织，观察心肌细胞超微结构，分别采用RT?PCR法和Western blot法检测心肌组织Nrf2、血红素加氧酶?l （HO?1）、醌氧化还原酶1（NQO1）、超氧化物歧化酶1（SOD1）及其mRNA的表达水平。结果与C组比较，再灌注末I∕R组和F组HR、＋dp∕dtmax和LVDP 降低，LVEDP 升高，EIP组LVDP降低，LVEDP升高（ P＜0．05），HR和＋dp∕dtmax差异无统计学意义（ P＞0．05）， I∕R组、EIP组和F组心肌组织Nrf2、HO?1、NQO1和SOD1及其mRNA的表达下调（ P＜0．05）；与I∕R组比较，EIP组和F组再灌注末HR、＋dp∕dtmax和LVDP 升高，LVEDP 降低，EIP 组心肌组织Nrf2、HO?1、NQO1和SOD1及其mRNA的表达上调， F组心肌组织Nrf2、HO?1、NQO1和SOD1的mRNA表达上调，Nrf2和HO?1的表达上调（ P＜0．05），NQO1和SOD1的表达差异无统计学意义（ P＞0．05）；与EIP组比较，F组再灌注末HR、＋dp∕dtmax和LVDP降低，LVEDP升高，心肌组织Nrf2、HO?1、NQO1和SOD1及其mRNA的表达下调（ P＜0．05）。结论乳化异氟醚后处理可能通过激活Nrf2?ARE信号通路，减轻大鼠心肌缺血再灌注损伤。
目的：評價乳化異氟醚後處理對大鼠缺血再灌註時覈轉錄因子 NF?E2相關因子2（ Nrf2）?抗氧化反應元件（ ARE）信號通路的影響。方法健康雄性SD大鼠，體重250～300 g，4～5月齡，採用Langendorff灌註裝置建立大鼠離體心髒灌註模型。取模型製備成功的心髒32箇，採用隨機數字錶法分為4組（ n＝8）：對照組（ C組）、缺血再灌註組（ I∕R組）、乳化異氟醚後處理組（ EIP組）和脂肪乳組（ F組）。平衡灌註20 min後，C組繼續灌註100 min；I∕R組32℃下缺血40 min，恢複灌註60 min；EIP組32℃下缺血40 min，于再灌註前即刻灌註含乳化異氟醚1．68 mmol∕L的K?H液2 min，繼續灌註37℃含氧的K?H液58 min；F組32℃下缺血40 min，于再灌註前即刻灌註含脂肪乳712 mg∕L 的K?H液2 min，繼續灌註37℃含氧K?H液58 min。分彆于平衡灌註末及再灌註末記錄HR、左室髮展壓（LVDP）、左室舒張末壓（LVEDP）和左心室壓力最大上升速度（＋dp∕dtmax）。于再灌註末，取左心室心肌組織，觀察心肌細胞超微結構，分彆採用RT?PCR法和Western blot法檢測心肌組織Nrf2、血紅素加氧酶?l （HO?1）、醌氧化還原酶1（NQO1）、超氧化物歧化酶1（SOD1）及其mRNA的錶達水平。結果與C組比較，再灌註末I∕R組和F組HR、＋dp∕dtmax和LVDP 降低，LVEDP 升高，EIP組LVDP降低，LVEDP升高（ P＜0．05），HR和＋dp∕dtmax差異無統計學意義（ P＞0．05）， I∕R組、EIP組和F組心肌組織Nrf2、HO?1、NQO1和SOD1及其mRNA的錶達下調（ P＜0．05）；與I∕R組比較，EIP組和F組再灌註末HR、＋dp∕dtmax和LVDP 升高，LVEDP 降低，EIP 組心肌組織Nrf2、HO?1、NQO1和SOD1及其mRNA的錶達上調， F組心肌組織Nrf2、HO?1、NQO1和SOD1的mRNA錶達上調，Nrf2和HO?1的錶達上調（ P＜0．05），NQO1和SOD1的錶達差異無統計學意義（ P＞0．05）；與EIP組比較，F組再灌註末HR、＋dp∕dtmax和LVDP降低，LVEDP升高，心肌組織Nrf2、HO?1、NQO1和SOD1及其mRNA的錶達下調（ P＜0．05）。結論乳化異氟醚後處理可能通過激活Nrf2?ARE信號通路，減輕大鼠心肌缺血再灌註損傷。
목적：평개유화이불미후처리대대서결혈재관주시핵전록인자 NF?E2상관인자2（ Nrf2）?항양화반응원건（ ARE）신호통로적영향。방법건강웅성SD대서，체중250～300 g，4～5월령，채용Langendorff관주장치건립대서리체심장관주모형。취모형제비성공적심장32개，채용수궤수자표법분위4조（ n＝8）：대조조（ C조）、결혈재관주조（ I∕R조）、유화이불미후처리조（ EIP조）화지방유조（ F조）。평형관주20 min후，C조계속관주100 min；I∕R조32℃하결혈40 min，회복관주60 min；EIP조32℃하결혈40 min，우재관주전즉각관주함유화이불미1．68 mmol∕L적K?H액2 min，계속관주37℃함양적K?H액58 min；F조32℃하결혈40 min，우재관주전즉각관주함지방유712 mg∕L 적K?H액2 min，계속관주37℃함양K?H액58 min。분별우평형관주말급재관주말기록HR、좌실발전압（LVDP）、좌실서장말압（LVEDP）화좌심실압력최대상승속도（＋dp∕dtmax）。우재관주말，취좌심실심기조직，관찰심기세포초미결구，분별채용RT?PCR법화Western blot법검측심기조직Nrf2、혈홍소가양매?l （HO?1）、곤양화환원매1（NQO1）、초양화물기화매1（SOD1）급기mRNA적표체수평。결과여C조비교，재관주말I∕R조화F조HR、＋dp∕dtmax화LVDP 강저，LVEDP 승고，EIP조LVDP강저，LVEDP승고（ P＜0．05），HR화＋dp∕dtmax차이무통계학의의（ P＞0．05）， I∕R조、EIP조화F조심기조직Nrf2、HO?1、NQO1화SOD1급기mRNA적표체하조（ P＜0．05）；여I∕R조비교，EIP조화F조재관주말HR、＋dp∕dtmax화LVDP 승고，LVEDP 강저，EIP 조심기조직Nrf2、HO?1、NQO1화SOD1급기mRNA적표체상조， F조심기조직Nrf2、HO?1、NQO1화SOD1적mRNA표체상조，Nrf2화HO?1적표체상조（ P＜0．05），NQO1화SOD1적표체차이무통계학의의（ P＞0．05）；여EIP조비교，F조재관주말HR、＋dp∕dtmax화LVDP강저，LVEDP승고，심기조직Nrf2、HO?1、NQO1화SOD1급기mRNA적표체하조（ P＜0．05）。결론유화이불미후처리가능통과격활Nrf2?ARE신호통로，감경대서심기결혈재관주손상。
Objective To evaluate the effect of emulsified isoflurane postconditioning on nuclear factor?E2 related factor 2 ( Nrf2 )?antioxidant response element ( ARE ) signaling pathway during myocardial ischemia?reperfusion ( I∕R ) in rats in vitro. Methods Healthy male Sprague?Dawley rats, aged 4-5 months, weighing 250-300 g, were heparinized and anesthetized with intraperitoneal 1% amobarbital sodium 40 mg∕kg. Their hearts were excised and perfused in a Langendorff apparatus with K?H solution. Thirty?two isolated rat hearts were randomly divided into 4 groups ( n=8 each ) using a random number table: control group (group C), group I∕R, emulsified isoflurane postconditioning group (EIP group) and fat emulsion group ( group F) . After 20 min of equilibration, group C was continuously perfused with K?H solusion for 100 min. Group I∕R underwent 40 min of ischemia at 32 ℃, followed by reperfusion for 60 min. In EIP and F groups, after undergoing 40 min of global ischemia, the isolated hearts were perfused for 2 min with K?H solution containing 1.68 mmol∕L emulsified isoflurane and 712 mg∕L intralipid, respectively, starting from the onset of reperfusion, and then were continuously perfused with K?H solution containing oxygen at 37 ℃ for 58 min. Heart rate (HR), left ventricular developed pressure (LVDP), left ventricular end?diastolic pressure ( LVEDP ) , and positive maximal pressure of left ventricular increase (+dp∕dtmax ) were recorded at the end of equilibration and reperfusion. At the end of reperfusion, myocardial specimens were obtained from the left ventricle for examination of the ultrastructure of myocardial cells and for determination of Nrf2, heme oxygenase?1 ( HO?1) , quinone oxidoreductase 1 ( NQO1) , and superoxide dismutase 1 ( SOD1) and mRNA expression using Western blot and real?time PCR. Results Compared with group C, HR, +dp∕dtmax and LVDP were significantly decreased, and LVEDP was increased at the end of reperfusion in I∕R and F groups, LVDP was significantly decreased, LVEDP was increased, and no significant changes were found in HR and +dp∕dtmax at the end of reperfusion in EIP group, and Nrf2, HO?1, NQO1 and SOD1 and mRNA expression was down?regulated at the end of reperfusion in I∕R, EIP and F groups. Compared with group I∕R, HR, +dp∕dtmax and LVDP were significantly increased, and LVEDP was decreased at the end of reperfusion in EIP and F groups, Nrf2, HO?1, NQO1 and SOD1 and mRNA expression was significantly up?regulated at the end of reperfusion in EIP group, and Nrf2, HO?1, NQO1 and SOD1 mRNA expression was significantly up?regulated, Nrf2 and HO?1 expression was up?regulated, and no significant changes were found in NQO1 and SOD1 expression at the end of reperfusion in group F. Compared with group EIP, HR, +dp∕dtmax and LVDP were significantly decreased, LVEDP was increased, and Nrf2, HO?1, NQO1 and SOD1 and mRNA expression was down?regulated in group F. Conclusion Emulsified isoflurane postconditioning attenuates myocardial I∕R injury probably by activating Nrf2?ARE signaling pathway in isolated rat hearts.
