Wu Kun, Ma Long, Xu Ting, Qin Zhensheng, Xia Tianfang, Wang Yi, Yu Xiangyou, Pang Liqun
Department of General Surgery, Huai'an First People's Hospital, Nanjing Medical University Huai'an 223300, Jiangsu, China.
Department of Intensive Care Uunit, First Hospital Affiliated to Xinjiang Medical University Urumqi, 830054, Xinjiang, China.
Int J Clin Exp Med. 2015 Aug 15;8(8):13818-23. eCollection 2015.
This study aims to investigate the protective effects and mechanism of recombinant adenovirus Ad.VSG-hBCL-2 towards ischemia/reperfusion injury in rat liver graft. Recombinant adenovirus Ad.VSG-hBCL-2 was injected into the donor rat liver of the experiment group through the portal vein, the laparotomy was performed for liver 36 h later, and the liver was save in lactated Ringer's solution at 4°C for 4 h, "two-cuff method" was used to perform the orthotopic liver transplantation. The bile secretion situations of two groups were observed 6 h after the portal vein reflow; the recipient rats were killed to detect the plasma levels of AST, ALT and LDH. And the expressions of Bcl-2 and TNF-α in liver tissue, and TUNEL assay was used to detect the apoptosis of liver tissue cells, electron microscopy was used to observe the changes of subcellular structures of liver tissue. 6 h after the surgery, the immunohistochemistry and Western Blot test showed that the Bcl-2 expression in the liver of the experiment group significantly increased than the control group, the bile secretion increased, the levels of AST, ALT and LDH were significantly lower, and the TNF-α expression increased significantly. The changes of cellular morphology of the experiment group were milder, and the apoptotic index was significantly lower than the control group. The portal vein-transfected recombinant adenovirus Ad.VSG-hBCL-2 could be effectively expressed in rat liver, and the high expressed Bcl-2 could reduce the ischemia/reperfusion injury in the transplanted liver.
本研究旨在探讨重组腺病毒Ad.VSG-hBCL-2对大鼠肝移植缺血/再灌注损伤的保护作用及机制。将重组腺病毒Ad.VSG-hBCL-2经门静脉注入实验组供体大鼠肝脏,36 h后开腹取肝,将肝脏于4℃保存于乳酸林格液中4 h,采用“双袖套法”行原位肝移植。门静脉再通6 h后观察两组胆汁分泌情况;处死受体大鼠,检测血浆AST、ALT和LDH水平。检测肝组织中Bcl-2和TNF-α的表达,采用TUNEL法检测肝组织细胞凋亡情况,电镜观察肝组织亚细胞结构变化。术后6 h免疫组化及Western Blot检测显示,实验组肝脏中Bcl-2表达较对照组显著增加,胆汁分泌增多,AST、ALT和LDH水平显著降低,TNF-α表达显著增加。实验组细胞形态变化较轻,凋亡指数显著低于对照组。经门静脉转染的重组腺病毒Ad.VSG-hBCL-2能在大鼠肝脏中有效表达,高表达的Bcl-2可减轻移植肝的缺血/再灌注损伤。