Cai Yijing, Zou Zhuolin, Liu Liyuan, Chen Si, Chen Yi, Lin Zhuo, Shi Keqing, Xu Lanman, Chen Yongping
Department of Infection Diseases, The First Affiliated Hospital of Wenzhou Medical University Wenzhou, Zhejiang 325000, P.R. China ; Wenzhou Key Laboratory of Hepatology Wenzhou, Zhejiang 325000, P.R. China ; Hepatology Institute of Wenzhou Medical University Wenzhou, Zhejiang 325000, P.R. China.
Wenzhou Key Laboratory of Hepatology Wenzhou, Zhejiang 325000, P.R. China ; Hepatology Institute of Wenzhou Medical University Wenzhou, Zhejiang 325000, P.R. China.
Int J Clin Exp Pathol. 2015 Jan 1;8(1):107-16. eCollection 2015.
To investigate the protective effect of bone marrow-derived mesenchymal stem cells (BMSCs) transplantation on acute liver injury (ALI) rats.
BMSCs were extracted from rat bone marrow, cultured and expansion in vitro, and identified by flow cytometer. Rat model with acute liver injury was established by employing D-galactosamine and Lipopolysaccharide. Male rats were randomly divided into ALI model group and BMSCs transplantation group. Rats were sacrificed 24 h, 72 h and 120 h after BMSCs injection to determine alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in serum. Proliferating cell nuclear antigen (PCNA) immunohistochemistry staining and quantitative reverse transcription polymerase chain reaction (RT-PCR) of α-fetoprotein (AFP) and glypican-3 (GPC3) were performed to analysis proliferation. Terminal deoxynucleontidyl Transferase Biotin-dUTP Nick End Labeling (TUNEL) assays were used to analyze apoptosis and mitochondria-dependent-pathway related factors Bax and Bcl-2 were examined by Western blot.
Compared with the ALI model group, the BMSCs transplantation group presented the lower levels of ALT, AST, decreased Bax proteins expression, and increased Bcl-2 expression. The mRNA levels of AFP and GPC3 and expression of PCNA were significantly higher in BMSCs transplantation group.
BMSCs transplantation could significantly restore liver function. These effects were supposed to be mediated by suppressing hepatocyte apoptosis as well as promoting proliferation. Reduction of apoptosis seemed to correlate with mitochondria-dependent-pathway.
探讨骨髓间充质干细胞（BMSCs）移植对急性肝损伤（ALI）大鼠的保护作用。
从大鼠骨髓中提取BMSCs，体外培养扩增，并用流式细胞仪进行鉴定。采用D-半乳糖胺和脂多糖建立大鼠急性肝损伤模型。将雄性大鼠随机分为ALI模型组和BMSCs移植组。在注射BMSCs后24小时、72小时和120小时处死大鼠，测定血清中丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）水平。进行增殖细胞核抗原（PCNA）免疫组化染色以及甲胎蛋白（AFP）和磷脂酰肌醇蛋白聚糖-3（GPC3）的定量逆转录聚合酶链反应（RT-PCR）以分析增殖情况。采用末端脱氧核苷酸转移酶生物素-dUTP缺口末端标记（TUNEL）法分析细胞凋亡，并通过蛋白质免疫印迹法检测线粒体依赖途径相关因子Bax和Bcl-2。
与ALI模型组相比，BMSCs移植组的ALT、AST水平较低，Bax蛋白表达降低，Bcl-2表达增加。BMSCs移植组中AFP和GPC3的mRNA水平以及PCNA的表达显著更高。
BMSCs移植可显著恢复肝功能。这些作用可能是通过抑制肝细胞凋亡以及促进增殖来介导的。细胞凋亡的减少似乎与线粒体依赖途径相关。
