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目的探讨同种异体骨髓基质细胞(BMSCs)移植到D-氨基半乳糖(D-Gal)诱导的兔急性肝损伤模型的治疗效果。材料与方法BMSCs由雄性白兔的股骨和胫骨的骨髓中提取,分离、培养及传代3周并观察其形态学特征,然后直接肝脏注射移植到D-Gal诱导的同种雄性兔的急性肝损伤模型内。实验组(n=24):为直接肝脏内注射5 ml BMSCs(约2×107个);对照组(n=34):为直接肝脏内注射5 ml D-Hanks液。每天观察各组实验兔的行为变化、存活率、肝功能[丙氨酸氨基转移酶(ALT),谷草转氨酶(AST),白蛋白(ALB)和总胆红素(TBIL)]的改变;BMSCs移植后1、2、4周分别处死两组实验兔,观察肝脏组织病理学变化及肝脏坏死程度,参照评分标准。两位组织病理学医师双盲予以各实验动物的肝脏组织标本评分。结果实验组1周后的存活率高于对照组(95.8%vs70.6%,P<0.05),而1～2周后的存活率较对照组未见显著性改善(100%vs 82.4%,P>0.05)。肝功能的检测显示:与对照组比较,实验组在细胞移植后血清ALB明显升高(P<0.05),ALT和AST均有显著性降低(P<0.05),但两组的TBIL水平的变化并无统计学意义(P>0.05)。组织病理学评分的结果显示:与对照组比较,BMSCs移植到受损肝脏后可显著减轻肝脏坏死程度及刺激肝细胞再生进而发挥强大的抗坏死作用(P<0.001)。结论同种异体BMSCs移植可加速修复D-Gal诱导的兔严重急性肝损伤。BMSCs直接肝内注射移植为实验动物模型简单实用的移植方式。
Objective To investigate the therapeutic effect of allogeneic bone marrow stromal cells (BMSCs) transplanted into D-galactose-induced acute liver injury model in rabbits. MATERIALS AND METHODS BMSCs were isolated, cultured, and passaged from bone marrow of femur and tibia of male white rabbits for 3 weeks and their morphological characteristics were observed. Then, BMSCs were transplanted into D-Gal-induced acute liver injury in allo-rabbits Inside the model. In the experimental group (n = 24), 5 ml of BMSCs (about 2 × 107) were injected directly into the liver; and in the control group (n = 34), 5 ml of D-Hanks solution was injected directly into the liver. The changes of behavior, survival rate, hepatic function (ALT, AST, ALB and TBIL) in rabbits in each group were observed daily. BMSCs At 1, 2 and 4 weeks after transplantation, two groups of experimental rabbits were sacrificed to observe the histopathological changes of liver and the degree of liver necrosis with reference to the grading standards. Two histopathologists double-blind to each experimental animal liver tissue specimens score. Results The survival rate of experimental group after 1 week was higher than that of control group (95.8% vs 70.6%, P <0.05), while the survival rate after 1 ~ 2 weeks was not significantly improved compared with control group (100% vs 82.4% P> 0.05). The liver function tests showed that the serum ALB was significantly increased (P <0.05) and the ALT and AST were significantly decreased (P <0.05) in the experimental group compared with the control group, but the change of TBIL level There was no statistical significance (P> 0.05). The results of histopathological score showed that transplanted BMSCs to the damaged liver could significantly reduce the degree of liver necrosis and stimulate the regeneration of hepatocytes, which in turn exert a strong anti-necrosis effect (P <0.001) compared with the control group. Conclusion Allogeneic BMSCs transplantation can accelerate the repair of D-Gal-induced severe acute liver injury in rabbits. BMSCs transplanted directly into the liver for the experimental animal model simple and practical method of transplantation.