构建大鼠减体积肝移植模型的肝脏信号转导差异蛋白表达

摘要

背景：蛋白质组学是目前医学界比较热门的科学研究技术，已经在肝移植的相关研究中得到初步应用，但未曾报道有在大鼠减体积肝移植相关研究中的应用。目的：利用蛋白质组学相关技术探讨大鼠减体积肝移植后与肝脏信号转导蛋白相关的差异蛋白。方法：在成功建立大鼠减体积肝移植模型的基础上，分别于移植后1，3，7 d获取移植肝脏组织，然后与预先获取冻存的供体和受体肝脏组织采用固相pH梯度双向凝胶电泳技术，建立双向凝胶电泳图谱，再利用串联质谱(MS-MS)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：实验中以变化倍数大于10倍为标准进行差异蛋白点的选择，总共发现了72个差异点，最终鉴定到了功能比较明确的32个蛋白，其中有4个蛋白参与了信号转导的过程，其分布于肝移植后的第3天和第7天，占6%。结果显示在大鼠减体积肝移植模型的成功和稳定建立的基础上，利用蛋白组学的相关技术，研究了大鼠减体积肝移植后参与肝脏信号转导的差异蛋白，为下一步深入研究调控这些蛋白的 MicroRNA 提供了有力的前期基础研究数据。%BACKGROUND:The proteome is a highlight technology in medical research fields lately, and has been reported to be applied in basic research fields related to liver transplantation. However, it has not been heard that the proteome has been used in research related to reduced-size liver transplantation. OBJECTIVE: To study expression of hepatic differential proteins related to signal transduction using proteomics after reduced-size liver transplantation in rats. METHODS:On the basis of successful establishment of rat models of reduced-size liver transplantation, transplanted liver tissues were obtained at 1, 3 and 7 days after transplantation. Postoperative liver tissue and normal donor, receptor liver tissues were subjected to solid pH gradient two-dimensional gel electrophoresis. Two-dimensional gel electrophoresis patterns were set up. Differentialy expressed protein spots were identified using tandem mass spectrometry analysis and database. RESULTS AND CONCLUSION:Seventy-two differential protein stains were found taking 10 times measure. Finaly, 32 proteins with clear functions were identified. Of them, four proteins participated in signal transduction, and they distributed at 3 and 7 days after liver transplantation, accounting for 6%. Results verified that on the basis of successful and stable establishment of rat models of reduced-size liver transplantation, proteomics technology was utilized to study differential proteins involving in signal transduction after reduced-size liver transplantation, and this study provides data for further deep investigation of regulating MicroRNA of these proteins.