CAJ | 학술논문

目的探讨细胞外信号调节激酶1/2(extracellular signal-regulated kinases,ERK1/2)通路在内毒素休克大鼠肺脏内血红素氧合酶-1 (heme oxygenase-1,HO-1)表达过程中作用. 方法清洁级雄性SD大鼠48只,体重180 g～200 g,采用随机数字表法,随机分为4组(每组12只):假手术组(S组)、内毒素休克组(SS组)、内毒素休克+阻断剂组(SE组)和阻断剂组(E组).S组和SS组股静脉输注0.1ml二甲亚砜(DMSO),SE组和E组股静脉ERK1/2阻断剂PD98059 10 μmol/kg(溶于0.1 mlDMSO)；30 min后,S组和E组分别给予0.5 ml生理盐水,SS组和SE组分别给予脂多糖(lipopolysaccharide,LPS) 10 mg/kg(溶于0.5 ml生理盐水),连续监测平均动脉压(mean arterial pressure,MAP),2h内SS组和SE组MAP下降≥基础血压25％,认为模型制备成功.根据病理学切片、超氧化物歧化酶(superoxide dismutase,SOD)活性及丙二醛(malondialdehyde,MDA)含量评定肺损伤程度,应用荧光定量PCR技术和Western blot技术测定ERK1/2和HO-1的表达,判断ERK1/2通路在内毒素休克大鼠肺脏HO-1表达过程中的作用. 结果 SS组的病理学评分(4.5±2.1)、肺含水率(81±6)％、MDA含量(3.12±1.30) nmol/mg明显高于S组(2.0±1.0)、(78±5)％、(1.79±0.12) nmol/mg和E组(2.1±1.1)、(77±4)％、(1.83±0.20) nmol/mg(P＜0.05),但低于SE组(6.4±1.3)、(87±5)％、(4.62±0.92) nmol/mg(P＜0.05)；SS组SOD活性(20.2±2.4)NU/mg明显低于S组(30.9±2.9)NU/mg和E组(32.2±1.2) NU/mg(P＜0.05),但高于SE组(14.9±3.2) NU/mg (P＜0.05)；SS组ERK1/2蛋白(0.43 ±0.09)及p-ERK1/2蛋白(1.46±0.22)、HO-1mRNA(1.834±0.023)及HO-1蛋白(1.17±0.25)表达明显高于SE组、S组和E组(P＜0.05)；上述指标在S组和E组之间差异无统计学意义(P＞0.05). 结论内毒素休克大鼠给予ERK1/2通路的阻断剂后,肺损伤加重,其机制可能与ERK 1/2被阻断后HO-1表达减少有关.
목적 탐토세포외신호조절격매1/2(extracellular signal-regulated kinases,ERK1/2)통로재내독소휴극대서폐장내혈홍소양합매-1 (heme oxygenase-1,HO-1)표체과정중작용. 방법 청길급웅성SD대서48지,체중180 g～200 g,채용수궤수자표법,수궤분위4조(매조12지):가수술조(S조)、내독소휴극조(SS조)、내독소휴극+조단제조(SE조)화조단제조(E조).S조화SS조고정맥수주0.1ml이갑아풍(DMSO),SE조화E조고정맥ERK1/2조단제PD98059 10 μmol/kg(용우0.1 mlDMSO)；30 min후,S조화E조분별급여0.5 ml생리염수,SS조화SE조분별급여지다당(lipopolysaccharide,LPS) 10 mg/kg(용우0.5 ml생리염수),련속감측평균동맥압(mean arterial pressure,MAP),2h내SS조화SE조MAP하강≥기출혈압25％,인위모형제비성공.근거병이학절편、초양화물기화매(superoxide dismutase,SOD)활성급병이철(malondialdehyde,MDA)함량평정폐손상정도,응용형광정량PCR기술화Western blot기술측정ERK1/2화HO-1적표체,판단ERK1/2통로재내독소휴극대서폐장HO-1표체과정중적작용. 결과 SS조적병이학평분(4.5±2.1)、폐함수솔(81±6)％、MDA함량(3.12±1.30) nmol/mg명현고우S조(2.0±1.0)、(78±5)％、(1.79±0.12) nmol/mg화E조(2.1±1.1)、(77±4)％、(1.83±0.20) nmol/mg(P＜0.05),단저우SE조(6.4±1.3)、(87±5)％、(4.62±0.92) nmol/mg(P＜0.05)；SS조SOD활성(20.2±2.4)NU/mg명현저우S조(30.9±2.9)NU/mg화E조(32.2±1.2) NU/mg(P＜0.05),단고우SE조(14.9±3.2) NU/mg (P＜0.05)；SS조ERK1/2단백(0.43 ±0.09)급p-ERK1/2단백(1.46±0.22)、HO-1mRNA(1.834±0.023)급HO-1단백(1.17±0.25)표체명현고우SE조、S조화E조(P＜0.05)；상술지표재S조화E조지간차이무통계학의의(P＞0.05). 결론 내독소휴극대서급여ERK1/2통로적조단제후,폐손상가중,기궤제가능여ERK 1/2피조단후HO-1표체감소유관.
Objective To investigate the effects of extracellular signal-regulated kinases (ERK1/2) signaling pathway on the expression of heme oxygenase-1 (HO-1) in acute lung injury of endotoxic shock induced by lipopolysaccharide (LPS) in rats.Methods Forty-eight pathogen-free male SD rats weighing 180 g-200 g were randomly divided into 4 groups (n=12 each):sham operation group (group S),endotoxic shock group (group SS),endotoxic shock and PD98059 group (group SE) and PD98059 group (group E).Rats in group S and group SS were administrated with DMSO 0.1 ml intravenously.Rats in Group SE and group E were injected with the inhibitor of ERK1/2 (PD98059) in 0.1 ml DMSO intravenously.30 min later,rats in Group S and group E received 0.5 ml normal saline intravenously while rats in group SS and group SE received LPS 10 mg/kg in 0.5 ml normal saline intravenously.The invasive arterial pressure was monitored,if there was an initial 25％ decrease in mean arterial pressure,then the model can be put to use.The lung injury degree was judged by microscopic examination,moisture content,malondialdehyde (MDA) content and superoxide dismutase (SOD) activity.The ERK1/2 protein,p-ERK1/2 protein and HO-1 protein were measured by Western blot.HO-1 mRNA level were measured by RT-PCR.Results The grade of the pathology (4.5 ±2.1),MDA content (3.12±1.30) nmol/mg,moisture content(81±6)％ in group SS were significantly higher than that in group S (2.0±1.0),(78±5)％,(1.79±0.12) nmol/mgand group E (2.1±1.1),(77±4)％,(1.83±-0.20) nmol/mg,(P＜0.05).But significantly lower than that in group SE(6.4±1.3),(87±5)％,(4.62±0.92) nmol/mg (P＜0.05).The SOD activity in group SS (20.2±2.4) NU/mg were significantly lower than that in group S (30.9± 2.9) NU/mgand group E (32.2±1.2) NU/mg(P＜0.05),but significantly higher than that in group SE(14.9±3.2) NU/mg(P＜0.05).ERK1/2 protein (0.43±0.09) and p-ERK1/2 protein (1.46 ±0.22),HO-1mRNA (1.834± 0.023) and HO-1 protein (1.17± 0.25) in group SS were significantly higher than that in group S,group SE and group E (P＜0.05).And there were no significant difference in group S and group P among all the indexes (P＞0.05).Conclusions Using the inhibitor of ERK1/2 signaling pathway in endotoxic shock rats worsened lung injury,and its mechanism may be related to decreased HO-1 expression by inhibition of ERK1/2 signaling pathway.