中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (1): 103-107.doi: 10.3969/j.issn.1673-8225.2010.01.022

• 干细胞与中医药 • 上一篇    下一篇

黄芪注射液促进脐血干细胞向肝细胞分化
具有增强移植干细胞治疗大鼠肝衰竭的效果

张永红1，曾 妍2，唐晓鹏1   

  1. 1中南大学湘雅二医院感染科，中南大学肝病研究所，湖南省长沙市 410011；
    2湖南省劳动卫生职业病防治研究所附属医院，湖南省长沙市  410007
  • 出版日期:2010-01-04 发布日期:2010-01-04
  • 通讯作者: 唐晓鹏，博士，教授，中南大学湘雅二医院感染科，中南大学肝病研究所，湖南省长沙市 410011 tangxiaopeng2008@163.com
  • 作者简介:张永红★，女，1965年生，四川省广安市人，汉族，2004年中南大学湘雅医学院毕业，硕士，副教授，主要从事各种肝病方面的研究。 zhangyh_xy@163.com
  • 基金资助:

    国家自然科学基金(39870651)；
    湖南省自然科学基金(04JJ6048)。

Astragalus membranaceus injection promotes hepatocytic differentiation of umbilical cord blood stem cell: Enhancement of cell transplantation for the treatment of liver failure

Zhang Yong-hong1, Zeng Yan2, Tang Xiao-peng1   

  1. 1Department of Infection, Xiangya Second Hospital, Central South University, Institute of Hepatology of Central South University, Changsha   410011, Hunan Province, China;
    2Affiliated Hospital of Hunan Provincial Institute for Labor Hygiene and Occupational Diseases, Changsha   410007, Hunan Province, China
  • Online:2010-01-04 Published:2010-01-04
  • Contact: Tang Xiao-peng, Doctor, Professor, Department of Infection, Xiangya Second Hospital, Central South University, Institute of Hepatology of Central South University, Changsha 410011, Hunan Province, China tangxiaopeng2008@163.com
  • About author:Zhang Yong-hong★, Master, Associate professor, Department of Infection, Xiangya Second Hospital, Central South University, Institute of Hepatology of Central South University, Changsha 410011, Hunan Province, China zhangyh_xy@163.com
  • Supported by:

    the National Natural Science Foundation of China, No.39870651*;
    the Natural Science Foundation of Hunan Province, No.04JJ6048*  

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摘要：

背景：在脐血干细胞移植治疗肝功能衰竭研究中，文献报道黄芪制剂可刺激造血干/祖细胞的增殖。

目的：观察黄芪注射液在体内外促进脐血干细胞向肝细胞的分化，以及增强脐血干细胞移植治疗大鼠肝衰竭的效果。

方法：取传至三四代的人脐血干细胞，药物筛选实验设立4组，分别加入0，40，200，400 mg/L黄芪，筛选适宜脐血干细胞生长的黄芪浓度；干细胞分化实验设立2组，分别加入肝细胞生长因子10 μg/L、肝细胞生长因子10 μg/L+黄芪200 mg/L。采用腹腔内注射D-氨基半乳糖法制备大鼠肝衰竭模型，建模48 h后仍存活的大鼠随机分为6组：模型对照组、黄芪组、鼠外周血单个核细胞组、联合组、联合+环磷酰胺组、联合+地塞米松组。RT-PCR法检测大鼠肝脏组织甲胎蛋白及白蛋白mRNA的表达，观察相关肝功能指标的变化。

结果与结论：不同质量浓度的黄芪制剂对人脐血干细胞生长影响不同，200 mg/L黄芪最适宜细胞生长和增殖。与单纯加入肝细胞生长因子的人脐血干细胞比较，另加入200 mg/L黄芪的人脐血干细胞白蛋白免疫组化染色阳性细胞数明显增加(P < 0.05)。随干预时间的延长，联合组、联合+环磷酰胺组、联合+地塞米松组肝脏组织白蛋白mRNA的表达均强于鼠外周血单个核细胞组；而甲胎蛋白mRNA的表达在早期均强于鼠外周血单个核细胞组，在后期则表达消失。干预7 d后，联合组、联合+环磷酰胺组、联合+地塞米松组的丙氨酸氨基转移酶、天门冬氨酸氨基转移酶及总胆红素下降幅度均明显大于模型对照组、黄芪组、鼠外周血单个核细胞组(P < 0.05)，而此3组间各项指标的变化无明显差异(P > 0.05)。提示200 mg/L黄芪在体内及体外均可以促进人脐血干细胞生长并分化为肝细胞，减轻肝脏损害，增强脐血干细胞移植治疗肝衰竭的效果。

关键词: 黄芪, 细胞分化, 肝细胞, 人脐血干细胞, 干细胞

Abstract:

BACKGROUND: Studies of umbilical cord blood stem cells transplantation for liver function failure have demonstrated that Astragalus membranaceus preparation can stimulate hemopoietic stem/progenitor cell proliferation.

OBJECTIVE: To explore the effect of Astragalus membranaceus injection on the differentiation of umbilical cord blood stem cell into hepatocyte in vitro and in vivo for the treatment of liver failure. METHODS: The third and fourth passage of human umbilical cord blood stem cells (HUCBSCs) were collected. In drug screening test, there were 4 groups: cells were separately cultured with 0, 40, 200, and 400 mg/L Astragalus membranaceus injection to screen the appropriate for cell growth. In cell differentiation test, there were 2 groups: HUCBSCs were respectively cultured with hepatocyte growth factor (HGF, 10 μg/L), and HGF (10 μg/L) plus 200 mg/L Astragalus membranaceus injection. D-aminogalactose was intraperitoneally injected to establish a model of acute liver failure. Surviving model rats (48 hours) were randomly divided into six groups: model control, Astragalus membranaceus injection, rat peripheral blood mononuclear cells, combination, combination+Cytoxan, and combination+dexamethasone groups. The alpha fetoprotein mRNA and albumin mRNA expression was determined by RT-PCR, and liver function indexes were observed.

RESULTS AND CONCLUSION: Different mass concentration of Astragalus membranaceus injection displayed varied influence on HUCBSC proliferation: 200 mg/L was the best for HUCBSC proliferation. Compared with HUCBSC cultured with HGF alone, the number of albumen-positive cells in HUCBSCs cultured with 200 mg/L Astragalus membranaceus injection and HGF was greater (P < 0.05). Moreover, the expression of albumen mRNA in combination, combination+Cytoxan, and combination+dexamethasone groups was greater than rat peripheral blood mononuclear cells group, while alpha fetoprotein mRNA expression was only greater than rat peripheral blood mononuclear cells group in early stage. At 7 days of treatment, the values of alanine aminotransferase, aspartate amino transferase and total bilirubin were significantly greater in combination, combination+Cytoxan, and combination+dexamethasone groups compared with model control, Astragalus membranaceus alone and rat peripheral blood mononuclear cells groups (P < 0.05), but no differences were observed among model control, Astragalus membranaceus alone and rat peripheral blood mononuclear cells groups (P > 0.05). Results indicate that Astragalus membranaceus injection at 200 mg/L can promote the proliferation and differentiation of HUCBSCs into hepatocyte in vitro and in vivo, ameliorate liver function and improve treatment effect of HUCBSC transplantation for liver failure.

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