摘要
目的：研究小鼠骨髓间充质干细胞（MSCs）和单个核细胞（MNCs）在四氯化碳（CCl4）诱导性大鼠肝损伤治疗中的作用。方法：无菌条件下取雄性Balb/c小鼠骨髓分离MNCs,培养纯化及流式细胞仪筛选获得MSCs;采用CCl4制作雌性Wistar大鼠肝损伤模型,随机分为MSCs移植组、MNCs移植组和肝损伤对照组。2个细胞移植组分别在CCl4损伤后立即经尾静脉注入小鼠MSCs或MNCs,各组分别于移植细胞输入前即刻（T0）、输入后第1（T1）、7（T2）、14（T3）、28（T4）、42（T5）天断尾取血检测丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）。6周后处死大鼠,比较肝脏质量指数,肝病理学切片检测肝损伤程度,PCR检测小鼠Sry基因片段植入大鼠肝脏的情况。结果：MSCsCD44、CD90均表达阳性,而CD45表达阴性;2个细胞移植组在T1~4时点ALT和AST浓度差异有统计学意义（P〈0.05）;与肝损伤对照组比较,MSCs移植组ALT和AST浓度在T1~4时点均降低;MSCs移植组肝损伤修复程度明显好于MNCs移植组和肝损伤对照组;仅MSCs移植组肝内有小鼠Y染色体阳性细胞存在。结论：体外分离培养及扩增的小鼠MSCs可植入到CCl4诱导性肝损伤大鼠的肝组织中,并改善CCl4导致的肝损伤。
Objective： To investigate the effect of mesenchymal stem cells （MSCs） and mononuclear cells （MNCs） from mice bone marrow on liver damage of carbon tetrachloride （CCl4）-treated rats. Methods：Bone marrow was collected from male Balb/c mice under sterile condition. MNCs were isolated, and MSCs were cultured, purified and screened by flow cytometry. Female Wistar rats were challenged with CCl4 to make liver damage models. The rats with liver damage were randomly divided into MSCs transplanted group, MNCs transplanted group and liver damage control group（n=10 in each group）. MSCs or MNCs were infused immediately through the tail veins after CCl4 treated in the cell transplanted rats. Tail blood samples of rats were collected for measuring the concentrations of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） before the induction of cell engraftment （T0）, and 1 d （T1）, 7 d （T2）, 14 d （T3）, 28 d （T4） and 42 d （T5） after the end of cell engraftment. After six weeks, the rat liver tissues were obtained to analyze the expression of Sry gene fragment with polymerase chain reaction. Results： According to flow cytometry assay, MSCs were positive for CD44 and CD90, while negative for CD45. There were significant differences in concentrations of ALT and AST at different time points between the two cell transplanted groups （P 0.05）. As compared with liver damage group, the concentrations of ALT and AST were significantly decreased at T1~4 in MSCs transplanted group, while not decreased at T1~4 in MNCs transplanted group. The histological examination suggested that liver damage recovery was much better only in these MSCs transplanted rats. PCR analysis revealed that mice Y-chromosome-positive cells can only be found in these MSCs transplanted rats. Conclusion： Mice MSCs could be isolated, cultured and expanded in vitro. Transplanted to CCl4-treated rats, they could engraft into the liver tissues of CCl4-treated rats and improve liver damage.
出处
《天津医药》
CAS
北大核心
2010年第6期509-512,547,共5页
Tianjin Medical Journal
基金
南昌大学科研基金(项目编号:JC44)
