Human adipose-derived mesenchymal stem cell transplantation via the tail vein for acute liver failure in rats

doi:10.3969/j.issn.2095-4344.1630

Abstract

Abstract:

BACKGROUND: Mesenchymal stem cells can protect and repair the liver of rats with liver failure, but the mechanisms are not completely clear.
OBJECTIVE: To explore the protective effects and related mechanisms of intravenous injection of human adipose-derived mesenchymal stem cells on acute liver failure in rats.
METHODS: Thirty-six Sprague-Dawley rats (provided by Qingdao Daren Fucheng Animal Husbandry Co., Ltd. in China) were randomly divided into control group, model group and transplantation group. Animal models of acute liver failure were established by intraperitoneal injection of D-galactosamine in the model group and the transplantation group. One day after modeling, the rats in the transplantation group were injected with human adipose-derived mesenchymal stem cell suspension, and those in the model group were injected with the same amount of saline. After 1 and 3 days of cell transplantation, the serum levels of alanine aminotransferase, aspartate aminotransferase, and total bilirubin were measured. Three days after cell transplantation, the serum levels of tumor necrosis factor-α, interleukin-6 and interleukin-10 were detected, the pathological changes of the rat liver were observed by hematoxylin-eosin staining, and the activity of glycogen synthase kinase-3β protein in the liver tissue was detected by western blot.
RESULTS AND CONCLUSION: Compared with the model group, there was a significant reduction in the serum levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, tumor necrosis factor-α, interleukin-6 and interleukin-10 in the transplantation group (P < 0.05). Inflammation and necrosis of liver tissues in the transplant group were alleviated compared with the model group. The activity of glycogen synthase kinase 3β in the liver tissue of the transplanted group was lower than that of the model group (P < 0.05). Overall, these results indicate that human adipose-derived mesenchymal stem cells can alleviate hepatic inflammation and pathological injury, and improve the liver function in rats with acute hepatic failure. Moreover, the mechanism may be related to the inhibition of glycogen synthase kinase 3β activity.

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: Adipose Tissue, Mesenchymal Stem Cell Transplantation, Liver Failure, Acute, Glycogen Synthase Kinase 3, Tissue Engineering

CLC Number:

Zhang Deguo, Tan Xueying, Tang Nan, Huang Fei, Shi Guangjun. Human adipose-derived mesenchymal stem cell transplantation via the tail vein for acute liver failure in rats[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(9): 1416-1421.

Figures/Tables 2

2.1 人脂肪间充质干细胞的培养及鉴定结果分离培养的人脂肪间充质干细胞原代接种于培养瓶，3 d后细胞呈短梭形；5 d后细胞数量增加，呈成纤维细胞样，散在生长；10 d后可见细胞融合达80%左右，呈漩涡状排列，见图1。流式细胞仪检测结果显示人脂肪间充质干细胞高表达CD29、CD49d、CD90，阳性率分别为99.7%，99.5%，99.1%；低表达CD34、CD45，阴性率分别为1.4%，0.1%，见图2。 2.2 实验动物数量分析每组12只大鼠参加实验，最终36只大鼠进入结果分析。 2.3 各组大鼠肝功能指标变化模型组大鼠血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶、总胆红素水平升高，与对照组相比差异有显著性意义(P < 0.05)。移植组大鼠血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶、总胆红素与对照组相比升高，但与同期模型组相比均有明显下降，差异有显著性意义(P < 0.05)，见图3。 2.4 各组大鼠血清炎性因子指标变化模型组大鼠血清肿瘤坏死因子α、白细胞介素6、白细胞介素10水平升高，与对照组相比差异有显著性意义(P < 0.05)。移植组大鼠血清炎性因子水平与模型组相比下降，差异有显著性意义(P < 0.05)，见图4。 2.5 各组肝脏病理组织学形态对照组肝脏颜色均匀，肝小叶结构完整清晰，未见坏死及炎性细胞浸润。模型组肝小叶结构破坏明显，肝细胞充血、水肿，可见肝细胞弥漫性坏死，大量炎性细胞浸润。移植组肝细胞坏死程度减轻，炎性细胞浸润减少，见图5。 2.6 各组大鼠肝脏组织糖原合成酶激酶3β表达糖原合成酶激酶3β第9位丝氨酸位点磷酸化越强，活性越弱。Western blot结果显示，模型组糖原合成酶激酶3β的磷酸化水平较对照组减弱，移植组较模型组的磷酸化水平升高，而总糖原合成酶激酶3β水平无明显改变，见图6。"

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