脐带间充质干细胞旁分泌物质对暴发性肝衰竭大鼠肝功能及肝细胞增殖的影响  

Effect of human umbilical cord mesenchymal stem cell-paracrine substance on liver function and hepatocytes proliferation in FHF rat

作　　者：孙琳琳[1] 朱争艳[2] 杜智 黎娇[2] 王鹏[2] 高英堂[4] 周佳美[1] 

机构地区：[1]天津医科大学三中心临床学院,300070 [2]天津市第三中心医院,天津市人工细胞重点实验室,再生医学与疾病生物治疗工程研究中心 [3]肝胆外科 [4]天津市肝胆疾病研究所

出　　处：《中华肝胆外科杂志》2011年第4期313-317,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：天津市发改委重点学科项目(再生医学与疾病生物治疗工程研究中心)资助

摘　　要：目的 探索人脐带间充质干细胞（human umbilical cord mesenchymal stem cells,UC-MSCs）旁分泌物质对实验性暴发性肝衰竭大鼠的治疗作用,研究其对大鼠肝功能及肝细胞增殖的影响.方法 体外分离培养人脐带间充质于细胞,流式细胞仪检测UC-MSCs的表面标志,制备含有UC-MSCs旁分泌物质的条件培养基（MSC-CM）,腹腔注射D-氨基半乳糖制备暴发性肝衰竭大鼠模型.实验分为3组：MSC-CM组、生理盐水（NS）组和促肝细胞生长素（PHGF）组.在模型制备后24 h从大鼠尾静脉分别注射三组治疗药物.每组8只大鼠治疗后12 h、24 h、36 h、60 h分别经内眦取血测定谷丙转氨酶（ALT）和总胆红素（TBIL）含量.每组另取5只大鼠在治疗后36 h取肝脏组织制备切片进行PCNA免疫组化染色,检测大鼠肝细胞增殖情况.观察并记录各组大鼠的生存状态及生存时间.结果 MSC-CM组及PHGF组大鼠治疗后24 h ALT及TBIL的含量均低于NS组（P＜0.05）,MSC-CM组与PHGF组比较差异无统计学意义.大鼠治疗后36 h肝脏切片PCNA染色显示,MSC-CM组和PHGF组PCNA肝细胞阳性数显著高于NS组（P＜0.01）,MSC-CM组与PHGF组比较差异无统计学意义.生存分析显示,MSC-CM组和PHGF组大鼠的生存率高于NS组（P=0.049）,MSC-CM组与PHGF组比较差异无统计学意义.结论 人脐带间充质干细胞的旁分泌物质可以刺激暴发性肝衰竭大鼠肝细胞增殖,改善暴发性肝衰竭大鼠的肝功能,为暴发性肝衰竭的治疗提供了一种新途径.Objective To investigate the therapeutic effect of human umbilical cord mesenchymal stem cell-paracrine substance on fulminant hepatic failure （FHF） rat, and to study the effect on liver function and hepatocyte proliferation. Methods Mesenchymal stem cells（MSCs）were separated from human umbilical cord, and surface makers of cells were detected by flow cytometry. Human umbilical cord mesenchymal stem cells-conditioned medium（MSC-CM） was prepared. FHF rat model was induced by intraperitoneal injection of D-galactosamine and they were randomly diveded into three groups： MSC-CM group, NS group, PHGF group. 24 h later, 1 ml MSC-CM, 1 ml 0. 9% NaCl solution and lml PHGF solution was injected into the tail vein of MSC-CM, NS, and PHGF rats, respectively. In each group （n=8 per group）, blood samples were collected at 12, 24, 36, and 60 h after treatment from inner canthus for analysis of blood ALT and TBIL levels. We used five rats per group for tissue collection after sacrifice at 36 h after treatment and 10 animals per group for survival analysis. PCNA immunohistochemical staining was used in the sections of liver tissue to detect hepatocyte proliferation. Results 24 h after treatment, the levels of ALT and TBIL in the MSC-CM and PHGF groups were lower than those in the NS group（P〈0. 05）, but there was no significant difference between the MSC-CM and PHGF groups. There were more PCNA-positive hepatocytes in the MSC-CM and PHGF groups than in the NS group（P〈0.01）, but there was no significant difference between MSC-CM and PHGF group. Survival analysis found that the survival rate of rats in the MSC-CM and PHGF groups was higher than that of rats in the NS group （P=0. 049）, but there was no significant difference between the MSC-CM and PHGF group. Conclusions The paracrine substance of human umbilical cord mesenchymal stem cells can stimulate hepatocyte proliferation and improve liver function of FHF rats, potentially creating a new avenue for the treatment of FHF.

关 键 词：脐带间充质干细胞 旁分泌 暴发性肝衰竭 肝功能 肝细胞增殖 

分 类 号：R575.3[医药卫生—消化系统]