中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2010年
2期
101-104
,共4页
牛铁生%齐国先%付鹏%孙英贤
牛鐵生%齊國先%付鵬%孫英賢
우철생%제국선%부붕%손영현
缺血预适应%缺氧诱导因子-1α%心肌梗死%缺血/再灌注损伤%心%蛋白激酶C
缺血預適應%缺氧誘導因子-1α%心肌梗死%缺血/再灌註損傷%心%蛋白激酶C
결혈예괄응%결양유도인자-1α%심기경사%결혈/재관주손상%심%단백격매C
Ischemic preconditioning%Hypoxia-indueihle factor-1α%Myocardial infarction%Myocardial ischemia/reperfusion injury%Protein kinase C
目的 观察缺氧诱导因子-1α(HIF-1α)对大鼠心肌缺血/再灌注损伤(IRI)的保护作用及蛋白激酶C(PKC)信号转导作用.方法 采用结扎冠状动脉左前降支30 min、再灌注180 min的方法建立IRI动物模型.将32只雄性Wistar大鼠按随机数字表法分为假手术组、IRI组、缺血预处理(IPC)组、IPC加PKC抑制剂组(IPC+Ⅰ组,IPC前静脉注射PKC抑制剂白屈菜季铵碱5 mg/kg),每组8只.再灌注180 min后取出心脏,测定HIF-1α、血红素氧合酶1(HO-1)的mRNA和蛋白以及天冬氨酸特异性半胱氨酸蛋白酶3(caspase-3)蛋白的表达;心内取血测定白细胞介素-8(IL-8)和髓过氧化物酶(MPO)水平.结果 与假手术组比较,IRI组心肌HIF-1α、HO-1的mRNA和蛋白及caspase-3蛋白表达均明显增多(HIF-1α mRNA:0.849±0.032比0.356±0.022,HIF-1α蛋白:0.762±0.042比0.324±0.016,HO-1 mRNA:0.862±0.045比0.332±0.012,HO-1蛋白:0.792±0.044比0.335±0.031,caspase-3蛋白:0.371±0.015比0.061±0.012,均P＜0.01),血IL-8、MPO显著升高[IL-8:(812±26)ng/L比(72±13)ng/L,MPO:(78.7±2.9)kU/L比(13.3±1.5)kU/L,均P＜0.01].与IRI组比较,IPC组HIF-1α、HO-1的mRNA和蛋白表达(HIF-1α mRNA,1.412±0.039,HIF-1α蛋白:1.362±0.045,HO-1 mRNA:1.523±0.038,HO-1蛋白:1.420±0.041)明显增多,caspase-3蛋白表达(0.129±0.019)明显降低(均P＜0.01),血IL-8[(432±59)ng/L]和MPO水平[(43.2±5.9)kU/L)明显降低(均P＜0.01).IPC+Ⅰ组各指标与IRI组无明显差异.结论 HIF-1α对心肌IRI具有重要保护作用,HIF-1α的表达是依赖PKC激活的,PKC是HIF-1α表达的重要信号转导通路.
目的 觀察缺氧誘導因子-1α(HIF-1α)對大鼠心肌缺血/再灌註損傷(IRI)的保護作用及蛋白激酶C(PKC)信號轉導作用.方法 採用結扎冠狀動脈左前降支30 min、再灌註180 min的方法建立IRI動物模型.將32隻雄性Wistar大鼠按隨機數字錶法分為假手術組、IRI組、缺血預處理(IPC)組、IPC加PKC抑製劑組(IPC+Ⅰ組,IPC前靜脈註射PKC抑製劑白屈菜季銨堿5 mg/kg),每組8隻.再灌註180 min後取齣心髒,測定HIF-1α、血紅素氧閤酶1(HO-1)的mRNA和蛋白以及天鼕氨痠特異性半胱氨痠蛋白酶3(caspase-3)蛋白的錶達;心內取血測定白細胞介素-8(IL-8)和髓過氧化物酶(MPO)水平.結果 與假手術組比較,IRI組心肌HIF-1α、HO-1的mRNA和蛋白及caspase-3蛋白錶達均明顯增多(HIF-1α mRNA:0.849±0.032比0.356±0.022,HIF-1α蛋白:0.762±0.042比0.324±0.016,HO-1 mRNA:0.862±0.045比0.332±0.012,HO-1蛋白:0.792±0.044比0.335±0.031,caspase-3蛋白:0.371±0.015比0.061±0.012,均P＜0.01),血IL-8、MPO顯著升高[IL-8:(812±26)ng/L比(72±13)ng/L,MPO:(78.7±2.9)kU/L比(13.3±1.5)kU/L,均P＜0.01].與IRI組比較,IPC組HIF-1α、HO-1的mRNA和蛋白錶達(HIF-1α mRNA,1.412±0.039,HIF-1α蛋白:1.362±0.045,HO-1 mRNA:1.523±0.038,HO-1蛋白:1.420±0.041)明顯增多,caspase-3蛋白錶達(0.129±0.019)明顯降低(均P＜0.01),血IL-8[(432±59)ng/L]和MPO水平[(43.2±5.9)kU/L)明顯降低(均P＜0.01).IPC+Ⅰ組各指標與IRI組無明顯差異.結論 HIF-1α對心肌IRI具有重要保護作用,HIF-1α的錶達是依賴PKC激活的,PKC是HIF-1α錶達的重要信號轉導通路.
목적 관찰결양유도인자-1α(HIF-1α)대대서심기결혈/재관주손상(IRI)적보호작용급단백격매C(PKC)신호전도작용.방법 채용결찰관상동맥좌전강지30 min、재관주180 min적방법건립IRI동물모형.장32지웅성Wistar대서안수궤수자표법분위가수술조、IRI조、결혈예처리(IPC)조、IPC가PKC억제제조(IPC+Ⅰ조,IPC전정맥주사PKC억제제백굴채계안감5 mg/kg),매조8지.재관주180 min후취출심장,측정HIF-1α、혈홍소양합매1(HO-1)적mRNA화단백이급천동안산특이성반광안산단백매3(caspase-3)단백적표체;심내취혈측정백세포개소-8(IL-8)화수과양화물매(MPO)수평.결과 여가수술조비교,IRI조심기HIF-1α、HO-1적mRNA화단백급caspase-3단백표체균명현증다(HIF-1α mRNA:0.849±0.032비0.356±0.022,HIF-1α단백:0.762±0.042비0.324±0.016,HO-1 mRNA:0.862±0.045비0.332±0.012,HO-1단백:0.792±0.044비0.335±0.031,caspase-3단백:0.371±0.015비0.061±0.012,균P＜0.01),혈IL-8、MPO현저승고[IL-8:(812±26)ng/L비(72±13)ng/L,MPO:(78.7±2.9)kU/L비(13.3±1.5)kU/L,균P＜0.01].여IRI조비교,IPC조HIF-1α、HO-1적mRNA화단백표체(HIF-1α mRNA,1.412±0.039,HIF-1α단백:1.362±0.045,HO-1 mRNA:1.523±0.038,HO-1단백:1.420±0.041)명현증다,caspase-3단백표체(0.129±0.019)명현강저(균P＜0.01),혈IL-8[(432±59)ng/L]화MPO수평[(43.2±5.9)kU/L)명현강저(균P＜0.01).IPC+Ⅰ조각지표여IRI조무명현차이.결론 HIF-1α대심기IRI구유중요보호작용,HIF-1α적표체시의뢰PKC격활적,PKC시HIF-1α표체적중요신호전도통로.
Objective To study the expression of hypoxia-inducible factor-1α (HIF-1α) in a rat model of myocardial ischemia/reperfusiou injury (IRI) and the role of protein kinase C (PKC) in signal pathway.Methods A rat model of myocardial IRI was reproduced by 30 minutes of left anterior descending coronary artery (LCA) occlusion followed by 180 minutes of reperfusion.Thirty-two healthy male Wistar rats were randomly divided into four groups.The first group was ischemie preconditioning (IPC) group; the second group was simple IRI group; the third group was IPC plus PKC inhibitor group (IPC±Ⅰ group); the fourth group was the sham-operation group without ligation of LCA.Eight rats were used in each group.The heart was harvested 180 minutes post-reperfusion,the mRNA and protein expression of HIF-1αand hemeoxygenase-1 (HO-1) were assessed.Meanwhile,the protein expression of caspase-3 was assayed.Blood samples were obtained from heart to determine the levels of interleukin-8 (IL-8) and myeloperoxidase (MPO).Results The mRNA and protein expression of HIF-1αand HO-1 increased significantly in the IRI group compared with the sham-operation group,while the protein expression of caspase-3 increased significantly in the IRI group (HIF-1α mRNA :0.849±0.032 vs.0.356±0.022,HIF-1α protein:0.762±0.042 vs.0.324±0.016,HO-1 mRNA:0.862±0.045 vs.0.332±0.012,HO-1 protein:0.792±0.044 vs.0.335±0.031,caspase-3 protein:0.371±0.015 vs.0.061±0.012,respectively,all P＜0.01).The levels of IL-8 and MPO increased significantly in the IRI group[IL-8:(812±626) ng/L vs.(72±613) ng/L,MPO:(78.7±2.9) kU/L vs.(13.3±1.5) kU/L,both P＜0.01].The protein and mRNA expression of HLF-1αand HO-1 increased significantly in the IPC group compared with IRI group (HIF-1α mRNA :1.412±0.039,HIF-1α protein:1.362±0.045,HO-1 mRNA:1.523±0.038,HO-1 protein:1.420±0.041,respectively),meanwhile the protein expression of caspase-3 (0.129±0.019) decreased significantly in the IPC group (all P＜0.01).The levels of IL-8[(432±59) ng/L]and MPO[(43.2±5.9) kU/L]decreased significantly in the IPC group compared with IRI group (both P＜0.01).All above parameters showed no significant change between IPC+Ⅰ group and IRI group.ConclusionHIF-1α plays a protective role in myocardial IRI,PKC is an important signal pathway of HIF-1α gene expression in IRI.
