CAJ | 학술논문

目的探讨脂氧素A4(LXA4)类似物(LXA4-ME)对急性胰腺炎(AP)大鼠的保护作用及其作用机制.方法将120只SD大鼠按数字表法随机分为假手术组、急性胰腺炎组(AP组)及LXA4-ME治疗组(LXA4-ME组),各40只.逆胰胆管注射5％牛磺胆酸钠(1 ml/kg)建立AP模型.LXA4-ME组在造模后经尾静脉注射含LXA4-ME(87.5 μg/kg)的生理盐水1 ml.假手术组及AP组造模后经尾静脉注射等量生理盐水.于12、24 h观察大鼠腹水的性质、量及胰腺病理学变化.测定血浆淀粉酶(AMY)含量.检测各组胰腺髓过氧化物酶(MPO)活性及丙二醛含量.反转录(RT)-PCR法检测各组胰腺肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-10及细胞间黏附分子(ICAM)-1、E-选择素、核因子κB p65的mRNA表达水平.免疫组织化学法检测各组胰腺核因子κB p65蛋白的表达.结果 LXA4-ME组大鼠12、24h腹水量[(6.88±1.23)、(6.53±0.91)ml]及胰腺病理学评分(8.7±1.3、7.8±1.1)均明显低于AP组[(12.32±1.94)、(14.15±1.68)ml,11.3±1.5、11.7±0.8,均P＜0.01]；12、24 h胰腺MPO活性、丙二醛及AMY水平均显著低于AP组(均P＜0.01)；胰腺TNF-α、IL-1β、ICAM-1、E-选择素、核因子κB p65的mRNA表达水平也均显著低于AP组(均P＜0.01),而IL-10 mRNA表达水平均高于AP组(均P＜0.01)；12、24 h胰腺核因子κB p65蛋白表达阳性率(24.8％±3.0％、31.6％±3.0％)显著低于AP组相应时间点(45.3％±3.4％、48.1％±4.6％,均P＜0.01).结论 LXA4-ME对AP大鼠具有一定的保护作用,其保护机制可能与减少核因子κB的激活有关.
목적 탐토지양소A4(LXA4)유사물(LXA4-ME)대급성이선염(AP)대서적보호작용급기작용궤제.방법 장120지SD대서안수자표법수궤분위가수술조、급성이선염조(AP조)급LXA4-ME치료조(LXA4-ME조),각40지.역이담관주사5％우광담산납(1 ml/kg)건립AP모형.LXA4-ME조재조모후경미정맥주사함LXA4-ME(87.5 μg/kg)적생리염수1 ml.가수술조급AP조조모후경미정맥주사등량생리염수.우12、24 h관찰대서복수적성질、량급이선병이학변화.측정혈장정분매(AMY)함량.검측각조이선수과양화물매(MPO)활성급병이철함량.반전록(RT)-PCR법검측각조이선종류배사인자(TNF)-α、백세포개소(IL)-1β、IL-10급세포간점부분자(ICAM)-1、E-선택소、핵인자κB p65적mRNA표체수평.면역조직화학법검측각조이선핵인자κB p65단백적표체.결과 LXA4-ME조대서12、24h복수량[(6.88±1.23)、(6.53±0.91)ml]급이선병이학평분(8.7±1.3、7.8±1.1)균명현저우AP조[(12.32±1.94)、(14.15±1.68)ml,11.3±1.5、11.7±0.8,균P＜0.01]；12、24 h이선MPO활성、병이철급AMY수평균현저저우AP조(균P＜0.01)；이선TNF-α、IL-1β、ICAM-1、E-선택소、핵인자κB p65적mRNA표체수평야균현저저우AP조(균P＜0.01),이IL-10 mRNA표체수평균고우AP조(균P＜0.01)；12、24 h이선핵인자κB p65단백표체양성솔(24.8％±3.0％、31.6％±3.0％)현저저우AP조상응시간점(45.3％±3.4％、48.1％±4.6％,균P＜0.01).결론 LXA4-ME대AP대서구유일정적보호작용,기보호궤제가능여감소핵인자κB적격활유관.
Objective To explore the protective effects and possible mechanism of lipoxin A4-methyl ester (LXA4-ME) in rats with acute pancreatitis (AP).Methods A total of 120 male SD rats were randomly divided into 3 groups:Sham operation (n=40),AP (n=40) and LXA4-ME (n=40).Sham operation group received normal salinc aftcr sham operation.AP was induced by a retrograde infusion of 5％ sodium taurocholate into pancreatobiliary duct. AP group received normal saline after modeling. In the LXA4-ME group,LXA4-ME was administered (87.5 μg/kg) intraveneously after the onset of AP.The rats were sacrificed at 12 h and 24 h post-induction.Their serum levels of amylase were detected.The amount of ascites was calculated and histological changes of pancreas were observed.The activities of myeloperoxidase (MPO) and levels of malonaldehyde (MDA) in pancreas were determined.The mRNA levels of tumor necrosis factor (TNF)-α,interleukin (IL)-1β,IL-10,intercellular adhesion molecule (ICAM-1),E-selectin and nuclear factor (NF)-κB p65 in pancreas were measured by reverse transcription-polymerase chain reaction (RT-PCR) . The expression of NF-κB p65 protein was also measured by immunohistochemistry.Results Compared with the AP group,the pathological scores of the LXA4-ME group improved ( 12 h:8.7±1.3 vs 11.3±1.5,24 h:7.8±1.1 vs 11.7±0.8 ) and the amount of ascites was lower(12 h:(6.88±1.23) ml vs (12.32±1.94) ml,24 h:(6.53±0.91) ml vs (14.15±1.68)ml,all P＜0.01 ).The serum levels of amylase in the LXA4-ME group were significantly lower than those in the AP group respectively at 12 h and 24 h post-operation (all P＜0.01).The activity of MPO and the level of MDA in pancreas in the LXA4-ME group were significantly lower than those in the AP group ( all P＜0.01 ).The pancreatic expressions of TNF-α mRNA,IL-1β mRNA,ICAM-1 mRNA,E-selectin mRNA and NF-κB p65 mRNA at 12 h and 24 h decreased in the LXA4-ME group versus the AP group at the corresponding timepoints( all P＜0.01 )while the expression of IL-10 mRNA increased versus the AP group at the corresponding timepoints ( all P＜0.01 ).Compared with that in the AP group,the pancreatic expression of NF-κB p65 protein decreased in the LXA4-ME group (12 h:24.8％±3.0％ vs 45.3％±3.4％,24h:31.6％±3.0％ vs 48.1％±4.6％,both P＜0.01).Conclusion LXA4-ME exerts protective effects in AP rats.And its mechanism may be due to the suppression of NF-κB activation