骨髓动员与骨髓间充质干细胞移植促进极量肝切除大鼠肝再生作用的对照研究  

A comparison of effects of bone marrow stem cell mobilization and bone marrow mesenchymal stem cell transplantation on the recovery of function and regeneration of rat liver tissue after massive excision of liver

作　　者：李东良[1] 何秀华[1] 范敬静[1] 张世安[1] 曾芝雨 张志强[1] 阮梅[1] 

机构地区：[1]厦门大学附属东方医院(南京军区福州总医院)肝胆内科,福州350025

出　　处：《解放军医学杂志》2014年第8期595-600,共6页Medical Journal of Chinese People's Liberation Army

基　　金：福建省社会发展重点资助项目(2011Y0043);南京军区医学科技创新基金(2007534)~~

摘　　要：目的观察骨髓间充质干细胞(BMSCs)移植与骨髓动员干细胞对极量肝切除大鼠肝功能恢复及肝再生的促进作用,探讨其在极量肝切除术后辅助治疗中的应用价值。方法建立极量肝切除(约85%)大鼠模型,随机分为4组。肝切除组(对照组,n=15):仅行极量肝切除;粒细胞集落刺激因子(G-CSF)组(n=15):腹腔注射rhG-CSF 150μg/(kg·d),连续5d;BMSCs组(n=15):大鼠尾静脉注射1.5ml BMSCs悬液(含1.5×106个BMSCs);G-CSF+BMSCs组(n=14):大鼠尾静脉注射1.5ml BMSCs悬液(含1.5×106个BMSCs),同时给予rhG-CSF 150μg/(kg·d)腹腔注射,连续5d。采用流式细胞仪检测G-CSF动员后大鼠外周血CD34+细胞比例;分别于术后第3、9天采血检测丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和白蛋白(ALB)等肝功能指标;最后一次采血后处死大鼠取肝组织,常规HE染色观察肝组织病理变化,采用免疫组织化学方法观察肝内Ki-67及BrdU阳性细胞表达情况。结果经流式细胞仪鉴定所分离细胞为BMSCs。rhGCSF骨髓动员后外周血BMSCs明显升高,5d后达高峰。对照组CD34+细胞比例为0.004%,G-CSF组骨髓动员3、5、9d后CD34+细胞比例分别为0.009%、0.016%、0.019%。3个治疗组术后第9天ALB均升高,与对照组比较差异有统计学意义(P<0.05或P<0.01)。3个治疗组Ki-67阳性细胞数(分别为110±28、104±34、99±22/400倍视野)及BrdU阳性细胞数(分别为18±6、16±6、17±5/400倍视野)均高于对照组(分别为73±27、12±4/400倍视野),差异有统计学意义(P<0.01)。结论 G-CSF动员自体BMSCs及同种异体大鼠BMSCs移植均有促进极量肝切除大鼠肝再生和肝功能恢复的作用,但二者联合应用未显示出协同作用。Objective To observe the promotion effect of bone mesenchymal stem cells （BMSCs） transplantation and mobilized bone marrow stem cells on the recovery of liver function and liver tissue regeneration after massive partial hepatectomy in rats. Methods The rats were randomly divided into four groups after massive partial hepatectomy （about 85%）, namely massive hepatectomy group （control group, n=15, received massive hepatectomy only）, G-CSF group [n=15, received rhG-CSF 150 μ g/（kg.d） by intraperitoneal injection after the operation for 5 days], MSCs group [n=15, received 5ml suspension of MSCs （about 1.5× 106） by tail vein injection after the operation], and G-CSF±MSCs group [n=14, received 1.5ml suspension of MSCs （about 1.5 × 106） by tail vein injection and rhG-CSF 150μ g/（kg.d） by intraperitoneal injection for 5 days after the operation]. The CD34± cells in the peripheral blood of the rats were assessed by flow cytometry after G-CSF mobilization. Liver function tests including ALT, AST and ALB were performed on the 3rd and 9th day after surgery. All the rats were sacrificed and liver tissue was harvested for histopathological study. The expression of Ki-67 and BrdU positive cells in the liver were determined by immunohistochemistry. Results Flow cytometric results identified the isolated cells were MSCs. BMSCs in peripheral blood significantly increased in number after rhGCSF mobilization, and reaching the maximum number on the 5th day. After mobilization for 3, 5 and 9 days, the positive CD34 cells in G-CSF group were 0.009%, 0.016%, 0.019% respectively, which were higher than those in control group. The level of albumin was significantly elevated in three treatment groups compared with control group on the 9th day after the hepatectomy （P〈O.05 or P〈0.01）. The expression of Ki-67 （110.16 ± 27.64, 103.57 ± 33.90, 98.52 ± 21.87 vs 72.39 ± 27.04 cells） and BrdU positive cells （17.96 ± 5.57, 16.45 ± 5.75, 16.66 ± 5.11 vs 11.72 _± 3.83 cells） in three treatmen

关 键 词：干细胞移植 肝切除术 肝再生 间质干细胞 

分 类 号：R657.3[医药卫生—外科学]