不同途径移植骨髓干细胞治疗急性肝损伤小鼠模型的效果比较
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摘要
目的探讨4种途径移植骨髓干细胞治疗的急性肝损伤小鼠的肝脏迁移情况及肝损伤的修复情况。方法雄性BALB/c小鼠分为A、B、C、D、E、F 6组,每组10只,A、B、C、D为移植组,E组为骨髓干细胞供体组,F组为急性肝损伤模型组。用CCL4/2-乙酰氨基芴制备小鼠急性肝损伤模型,分离小鼠骨髓干细胞,用红色荧光染料PKH26标记后经门静脉(A组,n=10)、尾静脉(B组,n=10)、腹腔(C组,n=10)及脾内(D组,n=10)输入到急性肝损伤小鼠体内,2周后处死小鼠,血清检测肝功能(ALT、AST、Alb),肝组织病理观察骨髓干细胞向肝脏迁移的情况及肝损伤小鼠的肝脏修复情况。F组小鼠于第8天处死检测ALT、AST及Alb值。计量资料2组间比较采用t检验,多组间比较采用单因素方差分析。结果显微镜下4个移植组移植的细胞均迁移到肝脏且通过病理图片均可见新生的肝细胞; ALT、AST、Alb值A、B、C、D 4组分别与F组比较差异均有统计学意义(ALT:t值分别为2. 372、2. 473、2. 354、2. 383,P值均<0. 05; AST:t值分别为2. 534、2. 423、2. 437、2. 643,P值均<0. 05; Alb:t值分别为2. 336、2. 243、2. 373、2. 352,P值均<0. 05)。结论骨髓干细胞促进急性肝损伤小鼠肝脏的修复,其修复程度与移植途径无关。
Objective To investigate the migration of bone marrow stem cells( BMSCs) to the liver and liver repair in mice with acute liver injury treated with BMSC transplantation through four approaches. Methods Male BALB/c mice were divided into groups A,B,C,D,E,and F,with 10 mice in each group. Groups A,B,C,and D were treated by transplantation,group E was used as the donor of BMSCs,and group F was used as the model of acute liver injury. CCL4/2-AFF was used to establish the model of acute liver injury. Mouse BMSCs were isolated,labeled with the red fluorescent dye PKH26,and then transplanted into the mice with acute liver injury through the portal vein( group A),the tail vein( group B),the abdominal cavity( group C),and the spleen( group D). The mice were sacrificed 2 weeks later. Serum levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST),and albumin( Alb) were measured. The pathology of liver tissue was observed to evaluate the migration of BMSCs to the liver and the degree of liver repair. The mice in group F were sacrificed on day 8 to measure the levels of ALT,AST,and Alb. The t test was used for comparison of continuous data between two groups,and one-way analysis of variance was used for comparison of continuous data between multiple groups. Results In groups A,B,C,and D,transplanted BMSCs migrated to the liver under a microscope,and newly formed hepatocytes were observed on pathological images. There were significant differences in the levels of ALT,AST,and Alb between groups A,B,C,and D and group F( ALT: t = 2. 372,2. 473,2. 354,and 2. 383,all P < 0. 05;AST: t = 2. 534,2. 423,2. 437,and 2. 643,all P < 0. 05; Alb: t = 2. 336,2. 243,2. 373,and 2. 352,all P < 0. 05). Conclusion BMSCs can promote repair of the liver in mice with acute liver injury,and the degree of liver repair is not related to the transplantation approach.
Objective To investigate the migration of bone marrow stem cells( BMSCs) to the liver and liver repair in mice with acute liver injury treated with BMSC transplantation through four approaches. Methods Male BALB/c mice were divided into groups A,B,C,D,E,and F,with 10 mice in each group. Groups A,B,C,and D were treated by transplantation,group E was used as the donor of BMSCs,and group F was used as the model of acute liver injury. CCL4/2-AFF was used to establish the model of acute liver injury. Mouse BMSCs were isolated,labeled with the red fluorescent dye PKH26,and then transplanted into the mice with acute liver injury through the portal vein( group A),the tail vein( group B),the abdominal cavity( group C),and the spleen( group D). The mice were sacrificed 2 weeks later. Serum levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST),and albumin( Alb) were measured. The pathology of liver tissue was observed to evaluate the migration of BMSCs to the liver and the degree of liver repair. The mice in group F were sacrificed on day 8 to measure the levels of ALT,AST,and Alb. The t test was used for comparison of continuous data between two groups,and one-way analysis of variance was used for comparison of continuous data between multiple groups. Results In groups A,B,C,and D,transplanted BMSCs migrated to the liver under a microscope,and newly formed hepatocytes were observed on pathological images. There were significant differences in the levels of ALT,AST,and Alb between groups A,B,C,and D and group F( ALT: t = 2. 372,2. 473,2. 354,and 2. 383,all P < 0. 05;AST: t = 2. 534,2. 423,2. 437,and 2. 643,all P < 0. 05; Alb: t = 2. 336,2. 243,2. 373,and 2. 352,all P < 0. 05). Conclusion BMSCs can promote repair of the liver in mice with acute liver injury,and the degree of liver repair is not related to the transplantation approach.
引文
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[12]FISCHER UM,HARTING MT,JIMENEZ F,et al.Pulmonary passage is a major obstacle for intravenous stem cell delivery:The pulmonary first-pass effect[J].Stem cells Dev,2009,18(5):683-692.
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[15]ZHAO MZ,WANG DD,YU ZJ,et al.Promotive effect of hepatocellular injury-conditioned medium on hepatic differentiation of bone marrow mesenchymal stem cells in rats[J].J Jilin Univ:Med Edit,2016,42(6):1108-1115.(in Chinese)赵美子,王丹丹,虞振静,等.肝细胞损伤条件培养基对大鼠骨髓间充质干细胞肝向分化的促进作用[J].吉林大学学报:医学版,2016,42(6):1108-1115.
[16]WU LL,XU B,WEI JW,et al.Therapeutical effect of local transplantation of bone marrow mesenchymal stem cells mixed with fibrin gel on rats with liver injury[J].J Clin Hepatol,2018,34(8):1740-1744.(in Chinese)吴林岚,徐兵,魏建威,等.纤维蛋白凝胶混合骨髓间充质干细胞局部移植治疗对肝损伤大鼠模型的影响[J].临床肝胆病杂志,2018,34(8):1740-1744.
[17]LIU W,YU SY,YAN HZ,et al.Therapeutic effect of MSCs transplantation on rats with hepatic fibrosis[J].Chin J Med Offic,2017,45(9):903-907.(in Chinese)刘卫,余森源,严和中,等.骨髓间质干细胞移植对肝纤维化模型大鼠治疗效果[J].临床军医杂志,2017,45(9):903-907.
[2]BERADIS S,DWISTHI S,NAJIMI M,et al.Use of mesenchymal stem cells to treat liver fibrosis:Current situation and future prospects[J].World J Gastroenterol,2015,21(3):742-758.
[3]JORNS C,NOWAK G,NEMETH A,et al.De novo donorspecific HLA antibody formation in two patients with criglernajjar syndrome type I following human hepatocyte transplantation with partial hepatectomy preconditioning[J].Am JTransplant,2015,16(3):1021-1030.
[4]SUK KT,YOON JH,KIM MY,et al.Transplantation with autologous bone marrow-derived mesenchymal stem cells for alcoholic cirrhosis:Phase 2 trial[J].Hepatology,2016,64(6):2185-2187.
[5]JANG YO,KIM YJ,BAIK SK,et al.Histological improvement following administration of autologous bone marrow-derived mesenchymal stem cells for alcoholic cirrhosis:A pilot study[J].Liver Int,2014,34(1):33-41.
[6]PENG L,TANG L,YE J,et al.Efficacy of autologous transplantation of bone marrow derived mesenchymal stem cells in treatment of decompensated cirrhosis[J].Chin J Liver Dis:Electr Version,2017,9(1):83-86.(in Chinese)彭蕾,汤磊,叶珺,等.自体骨髓间充质干细胞移植治疗失代偿期肝硬化的疗效[J].中国肝脏病杂志:电子版,2017,9(1):83-86.
[7]WAN PX,WANG BW,WANG ZC.Importance of the stem cell microenvironment for ophthalmological cell-based therapy[J].World J Stem Cells,2015,7(2):448-460.
[8]CALVI LM,LINK DC.The hematopoietic stem cell niche in homeostasis and disease[J].Blood,2015,126(22):2443-2451.
[9]XUE G,LI JL,LIU JF,et al.In vivo tracking of PKH26-labeled human umbilical cord mesenchymal stem cells after transplantation into rats with liver cirrhosis[J].Chin J Hepatol,2014,22(12):910-914.(in Chinese)薛改,李建立,刘建芳,等.PKH26标记示踪人脐带间充质干细胞在肝硬化大鼠体内的迁移[J].中华肝脏病杂志,2014,22(12):910-914.
[10]ZHANG LX,LI Y,WANG LM,et al.Bone marrow mesenchymal stem cell transplantation via different approaches in treatment of liver cirrhosis in mice[J].J Clin Hepatol,2016,32(10):1906-1910.(in Chinese)张丽霞,李莹,王黎明,等.不同途径移植骨髓间充质干细胞治疗肝硬化小鼠的效果比较[J].临床肝胆病杂志,2016,32(10):1906-1910.
[11]SONG YM,LIAN CH,WU CS,et al.Effects of bone marrowderived mesenchymal stem cells transplanted via the portal vein or tail vein on liver injury in rats with liver cirrhosis[J].Exp Ther Med,2015,9(4):1292-1298.
[12]FISCHER UM,HARTING MT,JIMENEZ F,et al.Pulmonary passage is a major obstacle for intravenous stem cell delivery:The pulmonary first-pass effect[J].Stem cells Dev,2009,18(5):683-692.
[13]LI HB,FU HY,LU TY,et al.Important progress in liver transplantation in 2017[J].Ogran Transplantation,2018,9(1):41-50.(in Chinese)李海波,符洪源,陆桐宇,等.肝移植领域2017年度重要进展盘点[J].器官移植,2018,9(1):41-50.
[14]MIYAJIMA A,TANAKA M,ITOH T.Stem/progenitor cells in liver development,homeostasis,regeneration,and reprogramming[J].Cell Stem Cell,2014,14(5):561-574.
[15]ZHAO MZ,WANG DD,YU ZJ,et al.Promotive effect of hepatocellular injury-conditioned medium on hepatic differentiation of bone marrow mesenchymal stem cells in rats[J].J Jilin Univ:Med Edit,2016,42(6):1108-1115.(in Chinese)赵美子,王丹丹,虞振静,等.肝细胞损伤条件培养基对大鼠骨髓间充质干细胞肝向分化的促进作用[J].吉林大学学报:医学版,2016,42(6):1108-1115.
[16]WU LL,XU B,WEI JW,et al.Therapeutical effect of local transplantation of bone marrow mesenchymal stem cells mixed with fibrin gel on rats with liver injury[J].J Clin Hepatol,2018,34(8):1740-1744.(in Chinese)吴林岚,徐兵,魏建威,等.纤维蛋白凝胶混合骨髓间充质干细胞局部移植治疗对肝损伤大鼠模型的影响[J].临床肝胆病杂志,2018,34(8):1740-1744.
[17]LIU W,YU SY,YAN HZ,et al.Therapeutic effect of MSCs transplantation on rats with hepatic fibrosis[J].Chin J Med Offic,2017,45(9):903-907.(in Chinese)刘卫,余森源,严和中,等.骨髓间质干细胞移植对肝纤维化模型大鼠治疗效果[J].临床军医杂志,2017,45(9):903-907.
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