Human amniotic mesenchymal stem cell transplantation for the repair of liver ischemia-reperfusion injury

doi:10.3969/j.issn.2095-4344.2016.45.015

Abstract

Abstract:

BACKGROUND: Studies have shown that human amniotic mesenchymal stem cells can differentiate into hepatocyte-like cells, suggesting that human amniotic mesenchymal stem cell transplantation provides a new potential for the clinical treatment of liver diseases.
OBJECTIVE: To observe the effect of human amniotic mesenchymal stem cell transplantation on the repair of liver ischemia-reperfusion injury repair.
METHODS: Sixty Sprague-Dawley rats were randomized into stem cell transplantation, model and control groups. Animal models of liver ischemia-reperfusion injury were made in the rats in the stem cell transplantation and model groups. One hour after modeling, rats in the stem cell transplantation were given injection of human amniotic mesenchymal stem cells (0.5 mL, 106 cells) via the tail vein, while rats in the model and control group were given L-DMEM (0.5 mL) or normal saline (0.5 mL), respectively. Liver function and liver morphology were detected at 1, 2, 3 weeks after transplantation. Meanwhile, RT-PCR detection and western blot assay were also conducted.
RESULTS AND CONCLUSION: (1) Liver function: Compared with the control group, levels of aspartate aminotransferase, alanine aminotransferase and malondialdehyde were significantly increased in the model group at different time points after transplantation (P < 0.05), while a significant reduction in the levels of these three indicators was found after cell transplantation as compared with the model group (P < 0.05). (2) Liver morphology: 2 weeks after transplantation, rats in the model group exhibited hepatocyte degeneration and necrosis, and severe fibrosis, but these changes were remarkably alleviated in the stem cell transplantation group. (3) PT-PCR and western blot detection: 2 weeks after transplantation, a significantly higher level of hepatocyte growth factor in the liver tissue and a lower level of α-smooth muscle protein were found in the stem cell transplantation group compared with the model group (P < 0.05). All these experimental findings indicate that human amniotic mesenchymal stem cell transplantation can improve impaired liver function in rats, possibly through regulating hepatocyte growth factor and α-smooth muscle protein expression levels in the liver, and thereby promotes the repair of liver ischemia-reperfusion injury.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Mesenchymal Stem Cell Transplantation, Reperfusion Injury, Tissue Engineering

CLC Number:

R394.2

Wang Xin-dang, Zhang Jian-jun, Ye Kui . Human amniotic mesenchymal stem cell transplantation for the repair of liver ischemia-reperfusion injury[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(45): 6788-6794.

Figures/Tables 2

2.1 hAD-MSCs的形态观察 hAD-MSCs在体外培养五六天可传代，传代后24 h内即完全贴壁，细胞呈梭形或多角形，增殖速度加快，培养至10 d左右时细胞生长面积可达80%-90%融合。传1代后的hAD-MSCs细胞生长迅速，呈长梭形集落生长，传第3 代hAD-MSCs细胞形态单一，呈漩涡状分布，细胞排列紧密，形态均匀，纯度较好，实验选取第3代hAD-MSCs细胞实施实验，第3代hAD-MSCs细胞见图1。 2.2 各组大鼠肝功能检测结果干细胞移植组与模型组大鼠均造模成功，进入结果分析。移植后1周，干细胞移植组、模型组门冬氨酸转移酶、丙氨酸转移酶和丙二醛明显高于正常对照组(P < 0.05)，干细胞移植组门冬氨酸转移酶、丙氨酸转移酶和丙二醛明显低于模型组(P < 0.05)。移植后2周，模型组门冬氨酸转移酶、丙氨酸转移酶和丙二醛明显高于正常对照组(P < 0.05)；干细胞移植组门冬氨酸转移酶、丙氨酸转移酶和丙二醛水平均明显低于模型组(P < 0.05)，但丙二醛水平与正常对照组比较仍较高(P < 0.05)。移植后3周，3组间门冬氨酸转移酶、丙氨酸转移酶和丙二醛水平两两比较差异均有显著性意义，见表1-3。 2.3 各组大鼠肝组织形态学变化移植后2周，模型组肝脏病理切片显示肝脏纤维化较严重，肝细胞出现水肿，脂肪样变性坏死，肝窦内出血，肝窦结构消失，假小叶形成明显，可见核固缩、溶解，炎性细胞浸润明显，肝小叶结构破坏较为严重，见图2A；干细胞移植组肝细胞坏死较少，亦可见假小叶形成，但数量较损伤组减少，肝脏水肿、纤维化、肝脂肪变性程度明显减轻，肝小叶结构明显恢复，见图2B；正常对照组肝脏结构完整清晰，质软、色红，无炎性细胞浸润及细胞水肿、变性、坏死等现象，见图2C。 2.4 各组肝细胞生长因子及α-平滑肌蛋白基因表达肝细胞生长因子主要由肝脏间质细胞产生；α-平滑肌蛋白主要分布于肝窦周围、汇管区、纤维间隔及增生的胆管周围细胞。移植后2周，RT-PCR法检测得知，与模型组比较，干细胞移植组中肝细胞生长因子基因表达水平明显升高(P < 0.05)，而α-平滑肌蛋白基因表达水平明显下降(P < 0.05)；模型组、干细胞移植组肝细胞生长因子、α-平滑肌蛋白基因表达与正常对照组比较差异有显著性意义(P < 0.05)，见图3。 2.5 各组肝细胞生长因子及α-平滑肌蛋白表达移植后2周，Western blot法检测得知，与模型组比较，干细胞移植组中肝细胞生长因子蛋白表达水平明显升高(P < 0.05)，α-平滑肌蛋白表达水平明显下降(P < 0.05)；模型组、干细胞移植组肝细胞生长因子蛋白、α-平滑肌蛋白与正常对照组比较差异有显著性意义(P < 0.05)，见图4。"

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