骨髓间充质干细胞异体移植对大鼠肝细胞增殖影响的实验研究

目的探讨骨髓间充质干细胞治疗急性肝衰竭的机制。方法全骨髓贴壁法培养扩增原代SD大鼠骨髓干细胞，传代培养，形态学、生长曲线及免疫表型鉴定骨髓间充质干细胞，移植治疗D-氨基半乳糖联合脂多糖致肝衰竭大鼠，观察干预治疗后1d、3d、5d、7d肝组织病理学改变、肝组织PCNA蛋白、CyclinDlmRNA表达量。结果原代培养大鼠骨髓间充质干细胞传代后，形态较均一，为梭形或纺锤形，高表达CD90（94．3％），CD29（99．4％），不表达CD11b／c（5．7％），CD45（3．2％）。骨髓间充质干细胞移植减轻肝脏组织病理损伤（P〈0．05）。BMSCs治疗组PCNA的蛋白表达高于对照组（P〈0．01）。干预后不同时间点治疗组细胞周期蛋白D1的mRNA表达高于对照组（1．182±0．248VS0．468±0．053、0．858±0．114vs0．218±0．178），差异有统计学意义（P〈0．01）。结论骨髓间充质干细胞移植减轻肝脏病理损伤，上调细胞周期蛋白DlmRNA的表达，促进肝再生，对急性肝衰竭有治疗作用。

An experimental study about the influence of bone marrow mesenchymal stem cells allotransplantation on hepatocyte proliferation in acute hepatic failure rat model

Objective To explore the mechanism of transplantation of bone marrow mesenchymal stem cells in the treatment of acute liver failure. Methods Cultured and amplified the rat bone marrow mesenchymal stem cells by adherent culture method from primary generation to fifth generation. Acute hepatic failure was induced by two injections of D- galactosamine and one injection of Lipopolysaccharide, delivered intraperitoneally with a 12-hour interval between injections. 1 ml BMSCs 1.4 × 10^7/kg or 0.9% NaC1 solution （control） was infused into caudal vein. PCNA in liver tissue was detected by immunohistochemistry. The level of CyclinD1 mRNA in liver tissue was observed. Results BMSCs in vitro culture were homogenous population and exhibited a spindle-shaped morphology. BMSCs expressed CD29 and CD90, didn＇ t express CDllb or CD45. BMSCs therapy inhibited liver enzyme release （P 〈 0.05）. BMSCs therapy improved gross and microscopic liver histopathology （P 〈 0.05）. BMSCs therapy enhanced liver regeneration （P 〈 0.05）. BMSCs therapy up-regulated the expression of CyclinD 1 mRNA （P 〈 0.05）. Conclusion BMSCs promote the regeneration of liver tissues. BMSCs transplantation has a therapy effect on acute liver failure.