摘要
目的:研究采用人脐血单个核细胞(humancord blood mononuclear cell,CB-MNC)治疗肝硬化大鼠后,其肝细胞中基因表达的变化.方法:首先分离CB-MNC,利用流式细胞术分析其CD34+细胞含量;然后利用四氯化碳(carbon tetrachloride,CCl4)联合乙醇方法建立大鼠肝硬化模型,将CB-MNC通过尾静脉注射对肝硬化大鼠进行治疗,通过血清学检测和组织学染色证明其治疗效果;提取肝脏RNA,进行表达谱芯片杂交并对结果进行统计分析.结果:血清学检测表明应用CCl4和乙醇造模后,谷草转氨酶和谷丙转氨酶显著升高(P<0.05).组织病理学染色结果表明大鼠肝硬化模型制作成功.与对照组相比,CB-MNC治疗可以显著地降低谷草转氨酶、谷丙转氨酶和转肽酶(P<0.05),且肝脏形态好转,肝细胞坏死和脂肪病变减少.基因芯片分析结果显示:与对照组相比,脐血单个核细胞调节了蛋白异源三聚化、氧化去甲基化、焦点黏连、白细胞跨内皮迁移、细胞外受体相互作用、补体凝血和P450对外源物代谢等相关基因.结论:CCl4和乙醇混合法可以成功建立大鼠肝硬化模型,而CB-MNC治疗可以显著地改善肝损伤.在CB-MNC治疗肝硬化逆转肝损伤的过程中,可能通过上调了补体凝血相关基因及过氧化物酶体增殖物激活受体信号通路,下调焦点黏连、白细胞跨内皮迁移和细胞外基质受体相互作用等相关基因发挥治疗作用.
AIM： To investigate the gene expression profile changes in liver cells of a rat model after human cord blood mononuclear cell（CB-MNC） therapy for hepatic cirrhosis.METHODS： CB-MNC was isolated from human cord blood,and the characterization of CD34＋ cells in CB-MNC was performed by flow cytometry analysis.Hepatic cirrhosis in SD rats was induced by subcutaneous injection of carbon tetrachloride and oral administration of alcohol.CB-MNC or PBS were transplanted by intravenous injection.Histo-pathological staining and serological testing were used to compare the morphology and liver func-tion between different groups.The gene expression alterations were compared between the PBS group and CB-MNC group by gene microarray analysis.RESULTS： Flow cytometric analysis showed the ratio of CD34＋ cells was 0.45% in CB-MNC.The results of serological assay and histopathol-ogy proved that transplantation of MNCs could improve the liver function in the animal model of hepatic cirrhosis.The levels of alanine aminotrans-ferase（ALT）,aspartate aminotransferase（AST） and glutamyl transferase（GGT） were reduced in the CB-MNC group（P〈0.05）.Gene microarray analy-sis showed that compared with the control group,CB-MNC therapy up-regulated the expression of genes related to complement,coagulation and peroxisome proliferator-activated receptor（PPAR）,and down-regulated the expression of genes re-lated to focal adhesion,leukocyte transendothelial migration and extracellular matrix（ECM）-receptor interaction.CONCLUSION： CB-MNC might improve the liver function by increasing the expression of genes related to complement,coagulation and PPAR,and decreasing the expression of genes related to focal adhesion,leukocyte transendo-thelial migration and ECM-receptor interaction in rats with hepatic cirrhosis.
出处
《世界华人消化杂志》
CAS
北大核心
2014年第23期3388-3395,共8页
World Chinese Journal of Digestology
基金
山东大学自主创新基金资助项目
No.2012ZD023
山东省科技攻关计划基金资助项目
No.2013GSF11812~~
关键词
人脐血单个核细胞
肝硬化
基因表达谱
细胞治疗
Human cord blood mononuclear cells
Hepatic cirrhosis
Gene microarray
Cell therapy
