大鼠骨髓细胞CD34^+β_2m^-CD90^+亚群肝内移植的研究

Research of transplantation of rat′s bone marrow cells subsets CD34^+β_2m^-CD90^+ into livers

目的观察骨髓CD34+β2m-CD90+细胞在受体大鼠纤维化肝内的定居、生长以及对受体肝的影响。方法将雄性肝纤维化大鼠的骨髓CD34+β2m-CD90+细胞用PKH26-GL标记后,从门静脉注射到预先经放疗的同种模型雌性大鼠肝内;PCR检测雌鼠血清Y染色体的表达,荧光显微镜检测PKH26-GL标记的骨髓CD34+β2m-CD90+细胞在受体肝内的分布;病理学检查移植骨髓CD34+β2m-CD90+细胞后大鼠肝纤维化组织形态学改变;免疫组化检测肝内白蛋白、α-SMA的表达,放免法测定受体血清层粘连蛋白(LN)、透明质酸(HA)的水平,斑点杂交检测肝脏Ⅰ、Ⅲ型前胶原mRNA的表达。结果在移植后2周内,雌性受体血清一直能检测到Y染色体特异的Sry基因表达,肝内有PKH26-GL红色荧光分布并有聚集成团趋势;受体肝纤维化减轻,间隔和胞周性纤维化较对照组改善;受体血清层粘连蛋白(LN)、透明质酸(HA)的水平下降(P<0.05),肝组织α-SMA、Ⅰ、Ⅲ型前胶原mRNA的表达下降(P<0.05),白蛋白的表达升高(P<0.05)。结论大鼠骨髓CD34+β2m-CD90+细胞自体移植入纤维化肝后,可能定居并有一定程度的增殖;大鼠移植CD34+β2m-CD90+细胞后,受体肝纤维化程度有减轻的趋势。

[Objective] To observe the inhabitation, proliferation of bone marrow CD34^＋β2m-CD90^＋ cells into recipients＇ livers and its influence. [Methods] We injected the bone marrow CD34^＋β2m-CD90^＋ cells marked by the red fluorescent dye （PHK26-GL） of the male rats with hepatic fibrosis through the portal vein. Then detected the expression of Y-chromosome in the blood of the female rats with PCR, the expression of distribution of CD34^＋β2m-CD90^＋ cells marked by PKH26-GL with fluorescent microscope, the hostomorphological change of rats＇ livers with pathological method. And the expression of albumin, α-SMA. Laminin（LN）. hyaluronic acid （HA） and pro-collagen Ⅰ, Ⅲ mRNA with dot blot hybridization. [Results] Y-chromosome＇s speeifu sry gent can be detected constantly 2 weeks after transplation. The red fluorescence of PKH26-GL distributed in the livers and had tendency to aggregate. The fibrosis of rats＇ livers has lessened and the expression of pro-collagen Ⅰ , Ⅲ mRNA has descended （P 〈0.05）. [Conclusion] The bone marrow CD34^＋β2m-CD90^＋ cells can inhabit and proliferate in the rats＇; the fibrosis of rats＇ livers alleviate when transplanted bone marrow CD34^＋β2m-CD90^＋ ceils.