CAJ | 학술논문

目的研究细胞外调节蛋白激酶(extracellular regulated kinase 1/2,ERK1/2)信号转导通路在大鼠急性坏死性胰腺炎(ANP)时的变化规律,探讨ERK1/2特异性抑制剂PD98059对ANP时胰腺的保护作用.方法以5%牛磺胆酸钠胰胆管逆行注射建立ANP模型,5只大鼠作为正常组,余75只大鼠按数字表法随机分为假手术组、ANP组及PD98059组,各25只.术后15 min、30 min、1 h、3 h和6 h分批处死动物,取血及胰腺组织.常规胰腺病理检查并评分;ELISA方法检测大鼠血清IL-1β、TNF-α水平;酶化学法检测胰腺组织内MPO活性;Western blotting法检测胰腺组织磷酸化ERK1/2的表达量.结果假手术组胰腺无明显病理改变;ANP组胰腺损伤明显,3 h的病理评分为9.9±0.4,PD98059组胰腺损伤减轻,3 h病理评分为4.0±0.4,与ANP组相差显著(P<0.05).假手术组、ANP组和PD98059组大鼠3 h时血清TNF-α水平分别为(65.8±20.5)pg/ml、(286.5±50.3)pg/ml、(180.4±32.9)pg/ml;IL-1β水平为(85.8±25.5)pg/ml、(293.8±46.3)pg/ml、(200.5±33.6)pg/ml;胰腺MPO活性为(0.19±0.02)U/g、(0.61±0.05)U/g、(0.52±0.03)U/g.ANP组和PD98059组均显著高于假手术组,而PD98059组又显著低于ANP组(P值均<0.05).正常大鼠胰腺磷酸化ERK1/2表达量为1100±141;ANP组15 min、30 min时磷酸化ERK1/2表达量分别为5300±486、5621±384,1 h开始下降,6 h时几乎与假手术组相似;PD98059组造模后15 min、30 min的磷酸化ERK1/2表达量分别为4200±370、3600±290,显著低于同时间点ANP组(P值均<0.01).结论 ERK1/2信号转导通路参与大鼠ANP发病机制,PD98059干预可减少IL-1β、TNF-α的产生,降低胰腺组织MPO活性,改善胰腺的病理损伤程度.
목적 연구세포외조절단백격매(extracellular regulated kinase 1/2,ERK1/2)신호전도통로재대서급성배사성이선염(ANP)시적변화규률,탐토ERK1/2특이성억제제PD98059대ANP시이선적보호작용.방법 이5%우광담산납이담관역행주사건립ANP모형,5지대서작위정상조,여75지대서안수자표법수궤분위가수술조、ANP조급PD98059조,각25지.술후15 min、30 min、1 h、3 h화6 h분비처사동물,취혈급이선조직.상규이선병리검사병평분;ELISA방법검측대서혈청IL-1β、TNF-α수평;매화학법검측이선조직내MPO활성;Western blotting법검측이선조직린산화ERK1/2적표체량.결과 가수술조이선무명현병리개변;ANP조이선손상명현,3 h적병리평분위9.9±0.4,PD98059조이선손상감경,3 h병리평분위4.0±0.4,여ANP조상차현저(P<0.05).가수술조、ANP조화PD98059조대서3 h시혈청TNF-α수평분별위(65.8±20.5)pg/ml、(286.5±50.3)pg/ml、(180.4±32.9)pg/ml;IL-1β수평위(85.8±25.5)pg/ml、(293.8±46.3)pg/ml、(200.5±33.6)pg/ml;이선MPO활성위(0.19±0.02)U/g、(0.61±0.05)U/g、(0.52±0.03)U/g.ANP조화PD98059조균현저고우가수술조,이PD98059조우현저저우ANP조(P치균<0.05).정상대서이선린산화ERK1/2표체량위1100±141;ANP조15 min、30 min시린산화ERK1/2표체량분별위5300±486、5621±384,1 h개시하강,6 h시궤호여가수술조상사;PD98059조조모후15 min、30 min적린산화ERK1/2표체량분별위4200±370、3600±290,현저저우동시간점ANP조(P치균<0.01).결론 ERK1/2신호전도통로삼여대서ANP발병궤제,PD98059간예가감소IL-1β、TNF-α적산생,강저이선조직MPO활성,개선이선적병리손상정도.
Objective To investigate the changes of extraceUular regulated kinase 1/2 (ERK1/2) phosphorylation and assess the effects of blocking the ERK1/2 phosphorylation on rats with acute necrotizing pancreatitis (ANP). Methods The ANP model was induced by retrograde injection of 5% sodium tanrocholate into the biliary and pancreatic duct. 5 rats were treated as normal control. Other 75 Sprague-Dawley (SD) rots were randomly divided into sham operations(SO) group (n =25), ANP group (n =25) and PD98059 group (n =25). The rats were sacrificed at 15 min, 30 min, 1 h, 3 h and 6 h after ANP induction, the blood and pancreatic sample were taken. Pathological changes of pancreas were observed with light microscope and scored. The serum level of TNF-α and IL-1β was determined by ELISA. MPO activities in pancreas were measured by enzyme chemistry assay. Western blotting was performed to determine the phosphorylations of ERK1/2 in the pancreas homogenates. Results There was no significant pathologic changes in rats of SO group;but significant injuries occurred in ANP group, the pathologic score at 3 h was 9.9 ± 0.4;the extent of injuries attenuated in PD98059 group, the pathologic score at 3 h was 4.0 ± 0.4 (P < 0.05). The serum levels of TNF-α at 3 h in SO, ANP and PD98059 groups were (65.8 ± 20.5) pg/ml, (286.5 ± 50.3) pg/ml, (180.4±32.9)pg/ml, respectively;the serum levels of IL-1β at 3 h in SO, ANP and PD98059groups were (85.8 ± 25.5) pg/ml, (293.8 ± 46.3) pg/ml, (200. 5 ± 33.6) pg/ml, respectively;MPOactivities in pancreas were (0. 19 ± 0.02)U/g, (0.61±0.05)U/g, (0.52±0.03) U/g, and the values in ANP and PD98059 groups were significantly higher than those in SO group, while the values in PD98059 group were significantly lower than those in ANP group (P < 0.01). The expression of ERK1/2 phosphorylation in normal pancreas was 1100 ± 141, the expressions of ERK1/2 phosphorylation in ANP group at 15 min, 30 min were 5300 ± 486, 5621 ± 384, respectively;the expressions began to decrease 1 h later and returned the similar level as SO group at 6 h;the expressions of ERK1/2 phosphorylation in PD98059 group at 15 min, 30 rain were 4200 ± 370, 3600 ± 290, respectively;which were signifieanfly lower than those in ANP group (all P value < 0. 01). Conclusions The ERK1/2 signal transduetion pathway plays an important role in the pathogenesis of ANP. Inhibition of ERK1/2 phosphorylation by PD98059 may decrease the production of IL-1β, TNF-α and pancreatic MPO, attenuate the extent of pancreatic pathologic injuries.