Chen Shenglin, Shao Cunhua, Dong Tianfu, Chai Hao, Xiong Xinkui, Sun Daoyi, Zhang Long, Yu Yue, Wang Ping, Cheng Feng
Department of Hepatobiliary Surgery Ward of General Surgery, The Affiliated Wuhu No. 2 People's Hospital of Wannan Medical College, Wuhu, Anhui Province, China.
Department of Hepatobiliary Surgery, Dongying People's Hospital, Dongying, Shandong Province, China.
PLoS One. 2014 Nov 6;9(11):e111425. doi: 10.1371/journal.pone.0111425. eCollection 2014.
Recent studies have demonstrated that transplantation of ATP7B-transduced hepatocytes ameliorates disease progression in LEC (Long-Evans Cinnamon) rats, a model of Wilson's disease (WD). However, the inability of transplanted cells to proliferate in a normal liver hampers long-term treatment. In the current study, we investigated whether transplantation of ATP7B-transduced bone marrow mesenchymal stem cells (BM-MSCs) could decrease copper overload in LEC rats.
The livers of LEC rats were preconditioned with radiation (RT) and/or ischemia-reperfusion (IRP) before portal vein infusion of ATP7B-transduced MSCs (MSCsATP7B). The volumes of MSCsATP7B or saline injected as controls were identical. The expression of ATP7B was analyzed by real-time quantitative polymerase chain reaction (RT-PCR) at 4, 12 and 24 weeks post-transplantation. MSCATP7B repopulation, liver copper concentrations, serum ceruloplasmin levels, and alanine transaminase (ALT) and aspartate transaminase (AST) levels were also analyzed at each time-point post-transplantation.
IRP-plus-RT preconditioning was the most effective strategy for enhancing the engraftment and repopulation of transplanted MSCsATP7B. This strategy resulted in higher ATP7B expression and serum ceruloplasmin, and lower copper concentration in this doubly preconditioned group compared with the saline control group, the IRP group, and the RT group at all three time-points post-transplantation (p<0.05 for all). Moreover, 24 weeks post-transplantation, the levels of ALT and AST in the IRP group, the RT group, and the IRP-plus-RT group were all significantly decreased compared to those of the saline group (p<0.05 compared with the IRP group and RT group, p<0.01 compared with IRP-plus-RT group); ALT and AST levels were significantly lower in the IRP-plus-RT group compared to either the IRP group or the RT group (p<0.01 and p<0.05. respectively).
These results demonstrate that transplantation of MSCsATP7B into IRP-plus-RT preconditioned LEC rats decreased copper overload and was associated with an increase in MSC engraftment and repopulation.
最近的研究表明，移植经ATP7B转导的肝细胞可改善威尔逊病（WD）模型——长 Evans 肉桂色（LEC）大鼠的疾病进展。然而，移植细胞在正常肝脏中无法增殖阻碍了长期治疗。在本研究中，我们调查了移植经ATP7B转导的骨髓间充质干细胞（BM-MSCs）是否能降低LEC大鼠的铜过载。
在门静脉输注经ATP7B转导的间充质干细胞（MSCsATP7B）之前，对LEC大鼠的肝脏进行辐射（RT）和/或缺血再灌注（IRP）预处理。作为对照注射的MSCsATP7B或生理盐水的体积相同。在移植后4周、12周和24周通过实时定量聚合酶链反应（RT-PCR）分析ATP7B的表达。在移植后的每个时间点还分析了MSCsATP7B的再填充、肝脏铜浓度、血清铜蓝蛋白水平以及丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）水平。
IRP加RT预处理是增强移植的MSCsATP7B植入和再填充的最有效策略。与生理盐水对照组、IRP组和RT组相比，在移植后的所有三个时间点，该策略导致双预处理组中ATP7B表达和血清铜蓝蛋白更高，铜浓度更低（所有p<0.05）。此外，移植后24周，与生理盐水组相比，IRP组、RT组和IRP加RT组的ALT和AST水平均显著降低（与IRP组和RT组相比p<0.05，与IRP加RT组相比p<0.01）；与IRP组或RT组相比，IRP加RT组的ALT和AST水平显著更低（分别为p<0.01和p<0.05）。
这些结果表明，将MSCsATP7B移植到经IRP加RT预处理的LEC大鼠中可降低铜过载，并与MSCs植入和再填充增加相关。
