间充质干细胞移植对大鼠移植肝脏缺血再灌注损伤的影响

目的 研究骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)移植促进大鼠减体积肝移植术后肝脏缺血再灌注损伤修复及再生的作用.方法 本研究①取同龄健康Wistar大鼠骨髓进行BMSCs体外培养,用第3代细胞制备悬液.②建立同龄健康Wistar大鼠50%减体积肝移植模型,分为实验组(BMSCs经门静脉植入)和对照组(生理盐水经门静脉注射)观察大鼠术后生存状态,术后不同时间留取血清和肝组织标本,采用免疫组化、流式细胞等方法进行检测.结果 成功体外分离培养BMSCs和建立50%减体积肝移植模型;大鼠肝移植术后2 h,自由活动和饮水.实验组丙二醛(MDA)、超氧化物歧化酶(SOD)、丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)不同时间点与相应的对照组相比差异有统计学意义(均P＜0.05).实验组细胞周期:术后第2、3天时肝组织的G0/G1期细胞比例显著低于对照组(分别F=22.061,44.572,均P＜0.01).增殖细胞核抗原(PCNA)阳性细胞数增高,术后第2、3和7天时比较,差异有统计学意义(分别F=30.716,28.281,8.975,均P＜0.05).结论 移植肝局部植入BMSCs后,可以促进大鼠肝细胞增殖和加快减体积肝移植术后缺血再灌注损伤的修复.

Effects of syngraft bone marrow mesenchymal stem cells on ischemic reperfusion injury of rat liver graft

Objective To study the effects of syngraft bone marrow mesenchymal stem cells (BMSCs) on the recovery of ischemic reperfusion injury related liver graft and graft regeneration in a reduced-size liver transplantation in rats.Methods BMSCs were isolated from the femur of Wistar rat by density gradient centrifugation,the third generation cells were prepared in suspension.50% reduced-size liver transplantation models (Wistar donor liver to Wistar recipient) were established,which were divided into the experimental group (1 ml BMSCs suspension was injected into the portal vein) and the control group (1 ml saline was injected into the portal vein).After liver transplantation the status of the recipients was observed.After transplantation,liver and serum were collected and analyzed by biochemistry,immunohistochemistry and flow cytometry.Results BMSCs were in a high purity and homogeneity.Rats were up and about and drinking 2 hrs after transplantation.The difference of MDA,SOD,ALT and AST between the experimental group and the control group was statistically significant (all P ＜ 0.05).The cell cycle in the experimental group showed that G0/G1 ratio on the 2~(nd) and 3~(rd) day after transplantation was lower than that of the control group (respectively F = 22.061,44.572,all P ＜ 0.01).The number of positive PCNA cells in experimental group was higher than that in control group.The difference between these two groups on the 2~(nd),3~(rd) and 7~(th) day was statistically significant (respectively F = 30.716,28.281,8.975,all P ＜ 0.05).Conclusion Syngraft BMSCs transfused into the portal vein of the liver graft promotes both the proliferation of liver ceils and the recovery of isehemic reperfusion injury related graft liver.