CAJ | 학술논문

目的观察脑死亡对猪肝脏功能、形态的影响并探讨核因子κB(nuclear factor κB,NF-κB)在脑死亡发生机制中的作用.方法健康长白猪12头,随机分为2组:对照组6头,仅行麻醉维持24 h;脑死亡组6头,建立猪脑死亡模型并维持脑死亡状态24 h.脑死亡后(对照组持续麻醉后)6、12、24 h取血标本及开腹取同一部位肝组织,检测血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸氨基转移酶(espartate aminotransferase,AST)水平;ELISA法检测血清白细胞介素1β(intedeukin-1β,IL-1β)水平;实时荧光定量PCR(real-time fluorescence quantitative polymerase chain reaction,real-time PCR)法检测肝组织NF-κB mRNA变化;免疫组织化学法观察肝组织NF-κB p65蛋白表达情况.结果脑死亡组长白猪各时间点血清ALT、AST、IL-1β水平、肝组织NF-κB mRNA转录水平及肝组织NF-κB p65蛋白表达水平均较对照组高,两者相比差异有统计学意义(P<0.05).脑死亡组脑死亡后12 h肝细胞轻度浊肿;24 h细胞水肿,肝细胞疏松化.结论脑死亡导致猪肝脏出现功能与形态学的损伤性变化.其机制可能是脑死亡引起肝细胞NF-κB mRNA 转录和蛋白翻译水平升高,进而导致机体炎症介质合成与释放.
목적 관찰뇌사망대저간장공능、형태적영향병탐토핵인자κB(nuclear factor κB,NF-κB)재뇌사망발생궤제중적작용.방법 건강장백저12두,수궤분위2조:대조조6두,부행마취유지24 h;뇌사망조6두,건립저뇌사망모형병유지뇌사망상태24 h.뇌사망후(대조조지속마취후)6、12、24 h취혈표본급개복취동일부위간조직,검측혈청중병안산안기전이매(alanine aminotransferase,ALT)、천동안산안기전이매(espartate aminotransferase,AST)수평;ELISA법검측혈청백세포개소1β(intedeukin-1β,IL-1β)수평;실시형광정량PCR(real-time fluorescence quantitative polymerase chain reaction,real-time PCR)법검측간조직NF-κB mRNA변화;면역조직화학법관찰간조직NF-κB p65단백표체정황.결과 뇌사망조장백저각시간점혈청ALT、AST、IL-1β수평、간조직NF-κB mRNA전록수평급간조직NF-κB p65단백표체수평균교대조조고,량자상비차이유통계학의의(P<0.05).뇌사망조뇌사망후12 h간세포경도탁종;24 h세포수종,간세포소송화.결론 뇌사망도치저간장출현공능여형태학적손상성변화.기궤제가능시뇌사망인기간세포NF-κB mRNA 전록화단백번역수평승고,진이도치궤체염증개질합성여석방.
Objective To observe how brain death affects the hepatic morphology and function of pigs and explore the roles of NF-κB. Methods Under general anaesthesia twelve healthy pigs were allocated randomly to two groups:control group(6 pigs),with non-inflacted Foley balloon catheter placed in the cerebral ventricle for 24 h,and brain death group,6 pigs,with estabhshment of brain death for 24 h.The serum and hepatic tissues in the same locus were taken at 6 h,12 h,and 24 h after the initial conformation of brain death.AST and ALT were determined by automatic biochemistry analyzer.IL-1βwas determined by ELISA.The NF-κB mRNA was determined by Real-time PCR and the NF-κB p65 by immunohistochemistry. Results The AST,ALT,IL-1β in serum,the NF-κB mRNA and the NF-κB p65 in hepatic tissues in brain death group were higher than those in control group and they all increased with the time(P<0.05).In brain death group,hepatocytes were edematous lightly after 12 hours,and the swelling progressively deteriorated after 24 hours,but there were no necrosis. Conclusion The activated NF-κB by brain death promoted the synthesis and release of inflammatory mediators,resulting in the hepatic dysfunction.