无白蛋白受体鼠中源于移植肝非实质细胞的肝细胞功能研究

Albumin-producing hepatocytes derived from liver nonparenchymal cells transplanted into the analbuminemic rats

目的研究F344大鼠来源的肝非实质细胞（LNPCs）能否驻存于γ射线全身照射的先天性无白蛋白大鼠（F344alb）的骨髓，进而转化为产白蛋白肝细胞。方法研究分为5组：无细胞移植和γ射线全身照射组（组Ⅰ，n=5）；接受γ射线全身照射，无细胞移植（组Ⅱ，n=5）；移植1×10^7 LNPCs，无γ射线全身照射（组Ⅲ，n=5）；γ射线全身照射后移植1×10^7 LNPCs（组Ⅳ，n=5）；γ射线全身照射后移植1×10^7骨髓细胞（BMCs，组Ⅴ，n=5），8周后，处死大鼠。肝脏切片行白蛋白免疫染色，检测呈多于6个白蛋白阳性簇状分布的肝细胞群中提取的DNA和血清中白蛋白的水平。结果（1）生存率：接受LNPCs移植的γ射线全身照射F344alb（组Ⅳ）平均存活率83．3％，而仅γ射线全身照射的F344alb存活率为20．0％（组Ⅱ），同时接受BMCs移植的γ射线全身照射F344alb平均存活率的的达100．0％（组Ⅴ）。（2）白蛋白阳性肝细胞呈多于6个的簇状分布仅发现于γ射线全身照射后接受LNPCs或BMCs移植的肝切片中（组Ⅳ和组Ⅴ）。（3）从F344alb受体（组Ⅳ和组Ⅴ）骨髓细胞及肝脏切片中微切的呈多于6个白蛋白阳性簇状分布的肝细胞群中提取的DNA中可检测到正常的白蛋白基因序列。（4）组Ⅳ和组Ⅴ血清中白蛋白水平显著升高。结论F344来源的肝非实质细胞能驻存于γ射线全身照射的F344alb的骨髓，进而在肝内转化为产白蛋白肝细胞。

Objective To investigate whether liver nonparenchymal cells （LNPCs） could harbor in the bone marrow cavity of the F344alb recipients with irradiation and further differentiate into albumin-producing hepatocytes in liver. Methods All F344alb were divided into 5 groups respectively shared sham-operation （group Ⅰ , n = 5 ） ; no transplantation, but accepting 7 irradiation （group Ⅱ , n = 5） ; being transplanted with 1 ×10^7 LNPCs without γ-irradiation （group Ⅲ , n = 5） ; being transplanted with 1×10^7 LNPCs after γ-irradiation （group Ⅳ, n = 5） ;transplanted with 1×10^7 BMCs after γ-irradiation （Group Ⅴ , n = 5）. Slices of the liver were stained by immunostaining with anti-rat albumin antibody. DNA extracted from BMCs as well as serum albumin levels of recipients were examined. Results （ 1 ） Survival rate： γ-irradiated F344alb had a mean 83.3 % survival rate when they were transplanted with LNPCs （group Ⅳ ）,at mean time all γ-irradiated rats without LNPCs or BMCs transplantation （group Ⅱ ） only had 20.0 % and those with BMCs transplantation had a 100.0 % （group Ⅴ ） survival rate,respectively. （2） Clusters consisting of more than 6 alb ＋ hepatocytes were only detected in the livers of recipientst hat received transplantation of LNPCs and BMCs after γ-irradiation （group Ⅳ and Ⅴ）. （3） The normal gene sequence of albumin was detected in BMCs of F344alb recipients and dissected alb ＋ clusters in F344alb tranaplanted with LNPCs and BMCs after γ-irradiation （group Ⅳ and V ）. （4） Serum albumin levels of F344alb recipients in group Ⅳ and group Ⅴ were significantly increased. Conclusion LNPCs could harbor in BMCs of the F344alb recipients with γ-irradiation and further differentiate into albumin-producing hepatocytes in the liver.