CAJ | 학술논문

背景：目前，蛋白质组学是一项比较成熟的科研技术，已经在肝移植基础研究领域中初步得到应用，但在大鼠减体积肝移植相关研究中的应用未曾报道。 　　目的：应用蛋白质组学相关技术探讨大鼠减体积肝移植后与肝脏能量代谢蛋白相关的差异蛋白。 　　方法：肝移植的供体是Lewis雌性大鼠，受体是Wistar雄性大鼠，供受体体质量差约20 g；移植肝质量/受体肝质量≈50%。采用改良减体积肝移植模型。获取供体肝组织过程中按照要求进行减体积，修整供体肝脏，将门静脉和肝下下腔静脉分别套入不同型号的套管以备套入受体的门静脉和肝下下腔静脉，胆道用细小支撑管植入供体的胆管中以备插入到受体的胆管中；最后进行受体的肝移植，受体先切除受体的原来肝脏组织，然后将准备好的供体肝脏植入受体中。造模后1，3和7 d获取移植肝脏组织，然后与预先获取冻存的供、受体肝脏组织应用蛋白组学技术建立双向凝胶电泳图谱，再利用串联质谱分析及数据库对差异表达的蛋白质点进行鉴定。 　　结果与结论：采用变化倍数大于10倍为标准进行差异蛋白点的选择，结果总共发现了72个差异点，通过鉴定，最终32个蛋白的功能比较明确，其中有ATP合成酶β亚基、电子转化黄素蛋白β多肽和质子转运ATP合成酶3个蛋白参与了细胞能量代谢的过程，其分布于肝移植后的第1和7天，占6%。
배경：목전，단백질조학시일항비교성숙적과연기술，이경재간이식기출연구영역중초보득도응용，단재대서감체적간이식상관연구중적응용미증보도。 　　목적：응용단백질조학상관기술탐토대서감체적간이식후여간장능량대사단백상관적차이단백。 　　방법：간이식적공체시Lewis자성대서，수체시Wistar웅성대서，공수체체질량차약20 g；이식간질량/수체간질량≈50%。채용개량감체적간이식모형。획취공체간조직과정중안조요구진행감체적，수정공체간장，장문정맥화간하하강정맥분별투입불동형호적투관이비투입수체적문정맥화간하하강정맥，담도용세소지탱관식입공체적담관중이비삽입도수체적담관중；최후진행수체적간이식，수체선절제수체적원래간장조직，연후장준비호적공체간장식입수체중。조모후1，3화7 d획취이식간장조직，연후여예선획취동존적공、수체간장조직응용단백조학기술건립쌍향응효전영도보，재이용천련질보분석급수거고대차이표체적단백질점진행감정。 　　결과여결론：채용변화배수대우10배위표준진행차이단백점적선택，결과총공발현료72개차이점，통과감정，최종32개단백적공능비교명학，기중유ATP합성매β아기、전자전화황소단백β다태화질자전운ATP합성매3개단백삼여료세포능량대사적과정，기분포우간이식후적제1화7천，점6%。
BACKGROUND:At present, the proteome is a mature technology that has been applied in basic research fields related to liver transplantation. But, it has been not reported in research related to reduced-size liver transplantation. OBJECTIVE:To explore the expression of differential proteins related to hepatic energy metabolism fol owing reduce-size liver transplantation in rats by using by proteomic technology. METHODS:The improved model of reduced-size liver transplantation was used in this experiment. The donor was health female Lewis rats and the recipient was male Wistar rats for liver transplantation. The difference between the donor and the recipient was about 20 g. The weight of donor liver/the weight of recipient donor was approximately equal to 50%. The donor liver tissue was harvested and trimmed to the required size. The portal vein and infrahepatic vena cava were cannulated, and the biliary tract was implanted into the donor bile duct for transplantation. Then the donor was transplanted into the recipient after the removal of original liver tissue. Hepatic specimens were harvested by 1, 3 and 7 days after reduced-size liver transplantation. Then, the harvested specimens were compared with the normal donor and recipient liver tissue that were previously harvested and frozen, to generate two-dimensional gel electrophoresis profile using proteome technology. Then tandem mass spectrometry and databases analysis were performed after two-dimensional electrophoresis for identifying differential protein stains. RESULTS AND CONCLUSION:In this experiment, 72 differential protein stains with over lo-fold changes were selected. After identification, 32 proteins showed clear functions, and among them three differential proteins (ATP synthase beta subunit, electron-transferring flavoprotein beta peptide and proton-transferring ATP synthase) were involved in the process of cel energy metabolism. The proteins were distributed on 1 and 7 days after reduce-size liver transplantation, accounting for 6%.