Theruvath Tom P, Zhong Zhi, Pediaditakis Peter, Ramshesh Venkat K, Currin Robert T, Tikunov Andrey, Holmuhamedov Ekhson, Lemasters John J
Center for Cell Death, Injury and Regeneration, Medical University of South Carolina, Charleston, SC 29425, USA.
Hepatology. 2008 Jan;47(1):236-46. doi: 10.1002/hep.21912.
Graft failure after liver transplantation may involve mitochondrial dysfunction. We examined whether prevention of mitochondrial injury would improve graft function. Orthotopic rat liver transplantation was performed after 18 hours' cold storage in University of Wisconsin solution and treatment with vehicle, minocycline, tetracycline, or N-methyl-4-isoleucine cyclosporin (NIM811) of explants and recipients. Serum alanine aminotransferase (ALT), necrosis, and apoptosis were assessed 6 hours after implantation. Mitochondrial polarization and cell viability were assessed by intravital microscopy. Respiration and the mitochondrial permeability transition (MPT) were assessed in isolated rat liver mitochondria. After transplantation with vehicle or tetracycline, ALT increased to 5242 U/L and 4373 U/L, respectively. Minocycline and NIM811 treatment decreased ALT to 2374 U/L and 2159 U/L, respectively (P < 0.01). Necrosis and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) also decreased from 21.4% and 21 cells/field, respectively, after vehicle to 10.1% and 6 cells/field after minocycline and to 8.7% and 5.2 cells/field after NIM811 (P < 0.05). Additionally, minocycline decreased caspase-3 activity in graft homogenates (P < 0.05). Long-term graft survival was 27% and 33%, respectively, after vehicle and tetracycline treatment, which increased to 60% and 70% after minocycline and NIM811 (P < 0.05). In isolated mitochondria, minocycline and NIM811 but not tetracycline blocked the MPT. Minocycline blocked the MPT by decreasing mitochondrial Ca(2+) uptake, whereas NIM811 blocks by interaction with cyclophilin D. Intravital microscopy showed that minocycline and NIM811 preserved mitochondrial polarization and cell viability after transplantation (P < 0.05).
Minocycline and NIM811 attenuated graft injury after rat liver transplantation and improved graft survival. Minocycline and/or NIM811 might be useful clinically in hepatic surgery and transplantation.
肝移植后的移植物功能衰竭可能涉及线粒体功能障碍。我们研究了预防线粒体损伤是否会改善移植物功能。在威斯康星大学溶液中冷藏18小时后，对供体肝和受体进行原位大鼠肝移植，并分别用赋形剂、米诺环素、四环素或N-甲基-4-异亮氨酸环孢素（NIM811）进行处理。移植后6小时评估血清丙氨酸氨基转移酶（ALT）、坏死和凋亡情况。通过活体显微镜评估线粒体极化和细胞活力。在分离的大鼠肝线粒体中评估呼吸作用和线粒体通透性转换（MPT）。用赋形剂或四环素处理后进行移植，ALT分别升至5242 U/L和4373 U/L。米诺环素和NIM811处理分别将ALT降至2374 U/L和2159 U/L（P<0.01）。坏死和末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）也分别从用赋形剂处理后的21.4%和21个细胞/视野，降至米诺环素处理后的10.1%和6个细胞/视野，以及NIM811处理后的8.7%和5.2个细胞/视野（P<0.05）。此外，米诺环素降低了移植物匀浆中的半胱天冬酶-3活性（P<0.05）。用赋形剂和四环素处理后的长期移植物存活率分别为27%和33%，米诺环素和NIM811处理后分别升至60%和70%（P<0.05）。在分离的线粒体中，米诺环素和NIM811而非四环素可阻断MPT。米诺环素通过减少线粒体对Ca(2+)的摄取来阻断MPT，而NIM811则通过与亲环蛋白D相互作用来阻断。活体显微镜检查显示，米诺环素和NIM811在移植后可保持线粒体极化和细胞活力（P<0.05）。
米诺环素和NIM811减轻了大鼠肝移植后的移植物损伤并提高了移植物存活率。米诺环素和/或NIM811在肝脏手术和移植中可能具有临床应用价值。
