CAJ | 학술논문

背景:研究表明,应用四氯化碳建造的重型肝衰竭小鼠模型中移植骨髓间充质干细胞后能够明显改善肝功能,促进肝再生.目的:观察骨髓间充质干细胞移植后动物模型肝功能及肝硬化程度等指标的变化情况.方法:采用密度梯度离心法和贴壁培养法获取兔骨髓间充质干细胞.取传代培养至3代的兔骨髓间充质干细胞,添加肝细胞生长因子和表皮生长因子,诱导21 d后进行BrdU标记.建立肝硬化兔模型,实验组兔将收集的BrdU标记过的肝样细胞悬液注入门静脉,对照组注入相同体积生理盐水.细胞移植后21 d麻醉下处死试验兔,观察肝硬化兔肝脏内分布及肝功能与对照组变化情况.结果与结论:兔骨髓间充质干细胞在体外经肝细胞生长因子、表皮生长因子的诱导条件下,于21 d发现细胞呈类圆形,细胞染色显示,甲胎蛋白、血清白蛋白和糖原表达均呈阳性表达.肝样细胞经门静脉移植后,实验组兔肝生化指标与对照组相比较显著改善(P<0.05).BrdU免疫组织化学染色显示:在汇管区和肝索肝窦内均可见BrdU阳性细胞分布.说明兔骨髓间充质干细胞能够向肝样细胞分化,经门静脉移植诱导后的肝样细胞可以改善肝硬化兔的肝功能,促进兔肝细胞再生.
배경:연구표명,응용사록화탄건조적중형간쇠갈소서모형중이식골수간충질간세포후능구명현개선간공능,촉진간재생.목적:관찰골수간충질간세포이식후동물모형간공능급간경화정도등지표적변화정황.방법:채용밀도제도리심법화첩벽배양법획취토골수간충질간세포.취전대배양지3대적토골수간충질간세포,첨가간세포생장인자화표피생장인자,유도21 d후진행BrdU표기.건립간경화토모형,실험조토장수집적BrdU표기과적간양세포현액주입문정맥,대조조주입상동체적생리염수.세포이식후21 d마취하처사시험토,관찰간경화토간장내분포급간공능여대조조변화정황.결과여결론:토골수간충질간세포재체외경간세포생장인자、표피생장인자적유도조건하,우21 d발현세포정류원형,세포염색현시,갑태단백、혈청백단백화당원표체균정양성표체.간양세포경문정맥이식후,실험조토간생화지표여대조조상비교현저개선(P<0.05).BrdU면역조직화학염색현시:재회관구화간색간두내균가견BrdU양성세포분포.설명토골수간충질간세포능구향간양세포분화,경문정맥이식유도후적간양세포가이개선간경화토적간공능,촉진토간세포재생.
BACKGROUND:Bone marrow mesenchymal stem cel s transplantation can significantly improve liver function and promote liver regeneration in a mouse model of severe liver failure induced by carbon tetrachloride. OBJECTIVE:To investigate liver function and hepatic cirrhosis degree after bone marrow mesenchymal stem cel s transplantation. METHODS:Bone marrow mesenchymal stem cel s were isolated by density gradient centrifugation method and adherent culture method. Passage 3 bone marrow mesenchymal stem cels were treated with hepatocyte growth factor and epidermal growth factor and labeled with BrdU after induced for 21 days. At the same time, animal models of liver cirrhosis were established. The BrdU-labeled hepatocyte-like cels suspension and normal saline were injected into rabbits in the experimental and control groups through the portal vein. The rabbits were sacrificed under anesthesia at 21 days after transplantation. The hepatic cirrhosis degree and liver function of rabbits with cirrhosis were investigated. RESULTS AND CONCLUSION:Morphological changes in bone marrow mesenchymal stem cel s were observed after in vitro induced with hepatocyte growth factor and epidermal growth factor for 21 days and cel morphology changed into round. Cel staining results showed positive expression of alpha-fetoprotein, serum albumin and glycogen. After transplantation of hepatocyte-like cel s through the portal vein, the liver biochemical indicators in the experimental group were significantly improved when compared with the control group (P<0. 05). BrdU immunohistochemical staining showed the BrdU-positive cel s were observed in the portal area and hepatic sinusoids. Rabbit bone marrow mesenchymal stem cel s can be induced to differentiate into hepatocyte-like cel s, and the induced cel s can improve rabbit liver function and promote hepatocyte regeneration.