中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2010年
1期
80-85
,共6页
王静%李东野%夏勇%罗园媛%陈丹%潘德锋%朱红%张卓琦%徐通达
王靜%李東野%夏勇%囉園媛%陳丹%潘德鋒%硃紅%張卓琦%徐通達
왕정%리동야%하용%라완원%진단%반덕봉%주홍%장탁기%서통체
基因%Akt1%再灌注损伤%线粒体通透性转换
基因%Akt1%再灌註損傷%線粒體通透性轉換
기인%Akt1%재관주손상%선립체통투성전환
Genes%Akt1%Reperfusion injury%Mitochondrial permeability transition
目的: 研究Akt1基因对缺血再灌注(I/R)损伤心肌细胞的保护作用,并探讨Akt1基因对心肌I/R损伤时线粒体通透性转换(MPT)功能的影响.方法: 使用结扎/松解左冠状动脉前降支法,结扎冠状动脉前降支30 min,再灌注120 min,建立大鼠在体心肌I/R损伤模型.40只雄性SD大鼠随机分为5组,每组8只:假手术对照组(control组)、心肌缺血/再灌注组(I/R组)、转Akt1基因+I/R组(Ad-gene组)、空载体+I/R组(Ad-blank组)、转Akt1基因+I/R+Akt抑制剂组(Ad-inhibitor组).Ad-gene组术前48 h开胸心肌内直接分点注射脂质体Akt1质粒复合物,control组和I/R组分别注射同等体积磷酸盐缓冲液(PBS),Ad-blank组注射等体积脂质体,Ad-inhibitor组注射等体积脂质体Akt1质粒LY294002混合物.观察转染Akt1基因对大鼠I/R心脏乳酸脱氢酶(LDH)和肌酸激酶(CK)释放、心肌细胞凋亡、Akt1表达、线粒体和胞浆内细胞色素C(Cyc-C)表达以及MPT的影响.结果: Control组可见Akt1表达,但明显低于其余各组;与control组相比较,其余各组心肌细胞凋亡指数(AI)、LDH和CK均增高;与control组相比较, 其余各组Cyc-C较胞浆表达增加而线粒体内表达减少. Ad-gene组Akt1蛋白表达较control组、I/R组、Ad-blank组及Ad-inhibitor组均增加;Ad-gene组AI、LDH和CK较I/R组、Ad-blank组及Ad-inhibitor组均显著减少,但仍高于control组;Ad-gene组 Cyc-C较I/R组、Ad-blank组以及Ad-inhibitor组胞浆内表达减少而线粒体内表达增加;Ad-gene组较IR组、Ad-blank组和Ad-inhibitor组在540 nm处吸光度值增高,但明显低于control组.I/R组、Ad-blank组以及Ad-inhibitor组Akt1、Cyc-C、AI、LDH和CK以及在540 nm处吸光度值比较均无显著差异.结论: 转染Akt1基因对I/R损伤心肌细胞具有保护作用,该作用可能与Akt1抑制I/R诱导的心肌线粒体膜通透性转换,从而保护线粒体的功能有关.
目的: 研究Akt1基因對缺血再灌註(I/R)損傷心肌細胞的保護作用,併探討Akt1基因對心肌I/R損傷時線粒體通透性轉換(MPT)功能的影響.方法: 使用結扎/鬆解左冠狀動脈前降支法,結扎冠狀動脈前降支30 min,再灌註120 min,建立大鼠在體心肌I/R損傷模型.40隻雄性SD大鼠隨機分為5組,每組8隻:假手術對照組(control組)、心肌缺血/再灌註組(I/R組)、轉Akt1基因+I/R組(Ad-gene組)、空載體+I/R組(Ad-blank組)、轉Akt1基因+I/R+Akt抑製劑組(Ad-inhibitor組).Ad-gene組術前48 h開胸心肌內直接分點註射脂質體Akt1質粒複閤物,control組和I/R組分彆註射同等體積燐痠鹽緩遲液(PBS),Ad-blank組註射等體積脂質體,Ad-inhibitor組註射等體積脂質體Akt1質粒LY294002混閤物.觀察轉染Akt1基因對大鼠I/R心髒乳痠脫氫酶(LDH)和肌痠激酶(CK)釋放、心肌細胞凋亡、Akt1錶達、線粒體和胞漿內細胞色素C(Cyc-C)錶達以及MPT的影響.結果: Control組可見Akt1錶達,但明顯低于其餘各組;與control組相比較,其餘各組心肌細胞凋亡指數(AI)、LDH和CK均增高;與control組相比較, 其餘各組Cyc-C較胞漿錶達增加而線粒體內錶達減少. Ad-gene組Akt1蛋白錶達較control組、I/R組、Ad-blank組及Ad-inhibitor組均增加;Ad-gene組AI、LDH和CK較I/R組、Ad-blank組及Ad-inhibitor組均顯著減少,但仍高于control組;Ad-gene組 Cyc-C較I/R組、Ad-blank組以及Ad-inhibitor組胞漿內錶達減少而線粒體內錶達增加;Ad-gene組較IR組、Ad-blank組和Ad-inhibitor組在540 nm處吸光度值增高,但明顯低于control組.I/R組、Ad-blank組以及Ad-inhibitor組Akt1、Cyc-C、AI、LDH和CK以及在540 nm處吸光度值比較均無顯著差異.結論: 轉染Akt1基因對I/R損傷心肌細胞具有保護作用,該作用可能與Akt1抑製I/R誘導的心肌線粒體膜通透性轉換,從而保護線粒體的功能有關.
목적: 연구Akt1기인대결혈재관주(I/R)손상심기세포적보호작용,병탐토Akt1기인대심기I/R손상시선립체통투성전환(MPT)공능적영향.방법: 사용결찰/송해좌관상동맥전강지법,결찰관상동맥전강지30 min,재관주120 min,건립대서재체심기I/R손상모형.40지웅성SD대서수궤분위5조,매조8지:가수술대조조(control조)、심기결혈/재관주조(I/R조)、전Akt1기인+I/R조(Ad-gene조)、공재체+I/R조(Ad-blank조)、전Akt1기인+I/R+Akt억제제조(Ad-inhibitor조).Ad-gene조술전48 h개흉심기내직접분점주사지질체Akt1질립복합물,control조화I/R조분별주사동등체적린산염완충액(PBS),Ad-blank조주사등체적지질체,Ad-inhibitor조주사등체적지질체Akt1질립LY294002혼합물.관찰전염Akt1기인대대서I/R심장유산탈경매(LDH)화기산격매(CK)석방、심기세포조망、Akt1표체、선립체화포장내세포색소C(Cyc-C)표체이급MPT적영향.결과: Control조가견Akt1표체,단명현저우기여각조;여control조상비교,기여각조심기세포조망지수(AI)、LDH화CK균증고;여control조상비교, 기여각조Cyc-C교포장표체증가이선립체내표체감소. Ad-gene조Akt1단백표체교control조、I/R조、Ad-blank조급Ad-inhibitor조균증가;Ad-gene조AI、LDH화CK교I/R조、Ad-blank조급Ad-inhibitor조균현저감소,단잉고우control조;Ad-gene조 Cyc-C교I/R조、Ad-blank조이급Ad-inhibitor조포장내표체감소이선립체내표체증가;Ad-gene조교IR조、Ad-blank조화Ad-inhibitor조재540 nm처흡광도치증고,단명현저우control조.I/R조、Ad-blank조이급Ad-inhibitor조Akt1、Cyc-C、AI、LDH화CK이급재540 nm처흡광도치비교균무현저차이.결론: 전염Akt1기인대I/R손상심기세포구유보호작용,해작용가능여Akt1억제I/R유도적심기선립체막통투성전환,종이보호선립체적공능유관.
AIM:To investigate the effects of Akt1 gene transfection into myocardium after ischemia-reperfusion (I/R) on mitochondrial permeability transition. METHODS:Forty adult male SD rats were divided randomly into five groups with 8 rats each:control group,I/R group,Ad-gene group,Ad-blank group and Ad-inhibitor group. The rats in Ad-gene group were injected with 30 μL Lipofectamine 2000 solution including Akt1 gene to the myocardium 48 h before ischemia while those in control group and I/R group were injected with PBS of the same volume. Rats in Ad-blank group were injected with Lipofectamine 2000 of the same volume into myocardium. In Ad-inhibitor group 30 μL Lipofectamine 2000 and gene complexes with LY294002 were injected. Hemodynamics,apoptotic index,the concentrations of lactate dehydrogenase,creatine kinase,the expression of Akt1,cytosolic,mitochondrial cytochrome C and MPT were also measured. RESULTS:The lowest level of Akt1 protein expression was observed in control group. The protein expression of Akt1 in Ad-gene group was higher than that in I/R group,Ad-blank group and Ad-inhibitor group. The AI,LDH and CK in Ad-gene group were significantly lower than those in other groups except control group. Transfection of Akt1 markedly reduced the loss of mitochondrial cytochome C after I/R injury. Ad-gene transfection led to a significant increase in absorbance at 540 nm compared to I/R group,Ad - black group and Ad-inhibitor group (P<0.05). CONCLUSION:Akt1 gene prevents myocardial apoptosis after I/R injury. Akt1 gene also inhibits the opening of mitochondria permeability transition and protects mitochondrial functions of myocardium in I/R injury.
