Transplantation of MSCs Overexpressing HGF into a Rat Model of Liver Fibrosis
Transplantation of MSCs Overexpressing HGF into a Rat Model of Liver Fibrosis
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DOI：
10.1007/s11307-015-0869-x
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发表时间：
2016-02-01
影响因子：
3.1
通讯作者：
Shan, Hong
中科院分区：
文献类型：
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作者：
Lai, Lisha;Chen, Junwei;Shan, Hong
目的：本研究旨在探讨过表达人肝细胞生长因子（human hepatocyte growth factor，HGF）对间充质干细胞（mesenchymal stem cells，MSCs）在大鼠肝纤维化再生中的作用，以及对MSCs在大鼠肝内的磁共振（magnetic resonance，MR）追踪。观察SPIO-HGF/MSCs的特性。在体外和体内进行普鲁士蓝染色用于铁评估。将SPIO-HGF/MSCs（A组）或SPIO-GFP/MSCs（B组）移植到肝纤维化大鼠模型中，并对大鼠肝脏进行MR成像。测量信噪比（SNR）和R2*（1/T2*）值。采用普鲁士蓝染色检测MSCs在体内的分布情况，并对肝脏进行Ki 67免疫组化染色。结果：HGF转染阳性率为93.17%，SPIO标记成功率为97.80 ± 1.06%。用SPIO标记MSC在体外不改变细胞增殖。肝T2* WI信号强度在细胞移植后第1天下降，第15天（A组）和第13天（B组）恢复至移植前水平。A组SNR显著低于B组（P= 0.006），R2* 值显著高于B组（P <0.001）。R2* 值与SNR呈显著负相关。体内普鲁士蓝染色阳性细胞数A组多于B组。A组Ki 67阳性率为16.11 ± 2.13%，血清ALT/HA水平降低。结论：HGF转染促进了MSCs在肝脏的定位，有助于肝脏修复。R2* 值可能是除了SNR之外的一个可行的指标来跟踪SPIO-MSC在肝脏中的移植。
Purpose: The aim of this study is to evaluate the effect of overexpressing human hepatocyte growth factor (HGF) for mesenchymal stem cells (MSCs) in liver fibrosis regeneration and magnetic resonance (MR) tracking of MSCs in rat liver.Procedures: MSCs were transfected with ad-HGF/ad-green fluorescent protein (GFP) and labeled with superparamagnetic iron oxide (SPIO). The characteristics of SPIO-HGF/MSCs were investigated. Prussian blue staining for iron assessment was conducted in vitro and in vivo. SPIO-HGF/MSCs (group A) or SPIO-GFP/MSCs (group B) were transplanted into a rat model of liver fibrosis, and MR imaging of the rat liver was performed. The signal to noise ratio (SNR) and R2* (1/T2*) value were measured. Prussian blue staining was performed to detect the in vivo distribution of MSCs, and liver Ki67 immunohistochemistry (IHC) staining was studied. The serum levels of HGF, alanine aminotransferase (ALT) and hyaluronic acid (HA) were determined.Results: The positive rate of HGF transfection was 93.17 % and the HGF/MSCs were labeled with SPIO successfully (97.80 +/- 1.06 %). Labeling of MSCs with SPIO did not alter cell proliferation in vitro. The signal intensity of liver T2* WI images decreased on day 1 after cell transplantation and recovered to pre-transplantation level on day 15 (group A) and day 13 (group B). The SNR of group A were significantly lower than that of group B (P= 0.006), and the R2* values of group A were significantly higher than those of group B (PG0.001). The R2* value had a significantly negative correlation with SNR. There were more Prussian blue-positive cells in of group A were more than in group B in vivo. The positive rate of Ki67 was 16.11 +/- 2.13 %, and the serum level of ALT/HA was decreased in group A.Conclusion: HGF transfection improved MSCs localization in the liver and aided liver repair. The R2* value might be a feasible index in addition to SNR to track the SPIO-MSC transplantation in the liver.