人羊膜间充质干细胞移植后慢性酒精性肝损伤大鼠血生化指标含量的变化
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Changes of blood biochemical indexes in the rats with chronic alcoholic liver injury after transplantation of human amniotic mesenchymal stem cells
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    摘要:

    目的 有研究表明人羊膜间充质干细胞(human amniotic mesenchymal stem cells,hAMSCs)具有多种分化和增殖功能,其可以诱导分化成肝样细胞,从而对肝损伤有一定的修复作用。方法 体外复苏培养人羊膜间充质干细胞,66只健康雌性Wistar大鼠,随机取22只,不做任何处理为正常对照组,余44只采用白酒灌胃30 d的方式建立大鼠慢性酒精性肝损伤模型,建模后将其随机分为模型组(尾静脉注射1 mL PBS),hAMSCs组(经尾静脉注入1 mL hAMSCs(2×106个),每组各22只。于移植后4周,采用全自动生化分析仪分别检测各组大鼠血清丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)、血清总胆红素(TBIL)、血清白蛋白(ALB)、血清总蛋白(Tp);同时检测检测各组大鼠肝脏超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)、过氧化氢酶(CAT)、以及丙二醛(MDA)、IL-4含量变化;免疫荧光显微镜观察PKH-26标记的hAMSCs分布情况,TUNEL法检测各组肝细胞凋亡情况。结果 各组大鼠血生化检测结果显示,与正常对照组比较,模型组中ALT、AST、TBIL、Tp含量均显著升高,而与模型组比较,hAMSCs移植组中ALT、AST、TBIL、Tp含量均显著下降(P<0.05),与正常对照组比较,模型组中ALB含量明显下降,而与模型组比较,hAMSCs移植组中ALB含量明显升高(P<0.05);与模型组比较,hAMSCs移植组SOD、GSH-PX、CAT含量均显著增高,MDA、IL-4含量均显著下降,与正常对照组比较,hAMSCs移植组SOD、GSH-PX、CAT含量均降低,MDA、IL-4含量均升高(P<0.05);移植组可见移植的PKH-26标记阳性的hAMSCs细胞分布于肝脏组织内,其余各组未见此类细胞分布(P<0.05);TUNEL法检测各组肝细胞凋亡可见凋亡细胞数,模型组最多(34.27±5.71),hAMSCs移植组次之(18.42±3.95),正常对照组未见凋亡细胞。结论 人羊膜间充质干细胞移植能够改善大鼠肝硬化的血生化指标水平,hAMSCs移植后可以减轻慢性酒精性肝损伤大鼠的肝脏细胞凋亡。

    Abstract:

    Objective Studies have suggested that human amniotic mesenchymal stem cells (hAMSCs) have multiple differentiation and proliferation functions, and can be induced to differentiate into hepatocyte-like cells to repair the liver injury.Methods Human amniotic mesenchymal stem cell swere prepared by resuscitation of frozen cells. Sixty-six healthy female Wistar rats were divided randomly into normal control group (n=22) without any treatment,and rat models of chronic alcoholic liver injury(n=44) treated by gastric gavage of alcohol for consecutive 30 days. After modeling,22 of those 44 rats were randomly divided into model group (tail vein injection of 1 mL PBS), ad another 22 rats as hAMSCs group which received tail vein injection of 1 mL (2×106) hAMSCs.Four weeks after the transplantation, the rat serum alanine aminotransferase (ALT),aspartate aminotransferase(AST), total bilirubin (TBIL), albumin (ALB), total protein (TP)were detected by a full automatic biochemical analyzer, and the catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase(GSH PX), malondialdehyde (MDA) and IL-4 content in the liver tissue were detected,the distribution of PKH-26 labeled hAMSCs was observed by immunofluorescence microscopy, and TUNEL staining was used to detect apoptosis in liver cells.Results The rat blood biochemical test showed that compared with normal control group, the serum content of ALT, AST, Tp and TBIL in the model group was significantly higher,compared with those of the model group; the serum contents of ALT, AST, TBIL and Tp in the hAMSCs transplantation group were significantly decreased(P<0.05), compared with hose of the normal control group. The content of ALB in the model group was significantly decreased, and the content of ALB in hAMSCs transplantation group was significantly higher (P<0.05)compared with those of the model group. The contents of SOD GSH- PX and CAT in the liver tissues were significantly increased in the hAMSCs transplantation group. The contents of MDA and IL-4 were decreased significantly, compared with those of the normal control group. The contents of SOD GSH- PX, CAT in the liver tissues were decreased in the hAMSCs transplantation group, and the content of MDA and IL-4 in the liver tissues were significantly increased(P<0.05). PKH-26-labeled positive hAMSCs cells were observed to be distributed in the liver tissue of the hAMSCs transplantation group but not in the liver tissue of other groups (P<0.05).TUNEL assay detected apoptotic cells in the liver of all model groups, with the highest number in the model group (34.27±571) and less in the hAMSCs transplantation group (18.42±3.95), but not found in the normal control group.Conclusions Human amniotic mesenchymal stem cell transplantation can improve the blood biochemical indexes of rats with cirrhosis, and hAMSCs can reduce the apoptosis in liver cells in rats with chronic alcoholic liver injury.

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韩海燕,宋文广,刘锦,崔云涛,薛占国.人羊膜间充质干细胞移植后慢性酒精性肝损伤大鼠血生化指标含量的变化[J].中国比较医学杂志,2017,27(2):49~54.

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  • 最后修改日期:2016-09-09
  • 在线发布日期: 2017-03-09
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