Therapeutical effect of local transplantation of bone marrow mesenchymal stem cells mixed with fibrin gel on rats with liver injury
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摘要:
目的探讨用骨髓间充质干细胞(MSCs)治疗肝损伤的方法。方法用冻融法从大鼠血浆或人血浆提取富含纤维蛋白原(FIG)的冷沉淀,用冷沉淀制成纤维蛋白凝胶(FG);检测冷沉淀中FIG、纤维连接蛋白(Fn)、PT、部分凝血活酶时间(APTT),第Ⅷ凝血因子(FⅧ)活性值。用大鼠的FG与MSCs混合组成FG-MSCs。体外培养FG-MSCs,采用二甲基噻唑蓝法检测MSCs增殖率。将FG-MSCs的成分Ⅰ(含FIG、MSCs)和成分Ⅱ(含凝血酶)交替注射到肝损伤大鼠肝实质组织内同一位点,一次注射量可分配到2个或更多位点,形成FG凝胶包埋MSCs。首次移植后第21和28天,同法做第2次和第3次注射。在第3次注射后7 d,称大鼠体质量;处死大鼠取血浆测ALT、TBil、Alb浓度;取肝脏称重,计算大鼠肝指数(平均肝质量/平均体质量)。两组间比较采用t检验;多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果人血浆冷沉淀中的FIG、FⅧ、Fn浓度均明显高于原血浆中的相应浓度(t值分别为9.669、12.317、9.043,P值均<0.01),但凝血活性(PT、APTT)则无明显差异...
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关键词:
- 肝损伤 /
- 纤维蛋白凝胶 /
- 间质干细胞移植 /
- 大鼠,Sprague-Dawley
Abstract:Objective To investigate the method for the treatment of liver injury using bone marrow mesenchymal stem cells ( MSCs) .Methods The cryoprecipitate ( CP) rich in fibrinogen ( FIG) was extracted from rat plasma or human plasma using the freeze-thaw method and was used to make fibrin gel ( FG) . FIG, fibronectin ( Fn) , prothrombin time ( PT) , activated partial thromboplastin time ( APTT) , and factor Ⅷ activity were measured for the CP. The rat FG was mixed with rat MSCs to make FG-MSCs. The FG-MSCs were cultured in vitro, and MTT assay was used to measure the proliferation rate of MSCs. The components Ⅰ ( containing FIG and MSCs) and Ⅱ ( containing thrombin) of FG-MSCs were injected alternately into the same site of liver parenchyma tissue of the rats with liver injury to form FG-embedded MSCs, and a single dose of injection could be assigned to two or more sites. Such injection was repeated on days 21 and 28 after the first transplantation. On day 7 after the third injection, the body weight of rats was measured; the rats were sacrificed and plasma was collected to measure the levels of alanine aminotransferase ( ALT) , total bilirubin ( TBil) , and albumin ( Alb) ; the liver was weighed to calculate liver index [ ( mean liver weight/mean body weight) × 100%]. The t-test was used for comparison between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. Results The CP of human plasma had significantly higher concentrations of FIG, Fn, and factor Ⅷ than the former plasma ( t = 9. 669, 12. 317 and 9. 043, all P < 0. 01) , while there were no significant differences in PT and APTT ( both P > 0. 05) . After in vitro culture for 4 days, FG-MSCs had a slightly higher proliferation rate than MSCs ( P > 0. 05) . After the injection of FG-MSCs or MSCs into the liver tissue of the rats with liver injury, there were significant reductions in the plasma levels of ALT and TBil ( both P < 0. 05) and a significant increase in the level of Alb ( P < 0. 05) . Conclusion The injection of FG-MSCs or MSCs into the liver parenchyma tissue can significantly improve the liver function of rats with liver injury, and FG-MSCs can also prevent bleeding after injection.
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Key words:
- liver injury /
- fibrin gel /
- mesenchymal stem cell transplantation /
- rats, Sprague-Dawley
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[1]PEI YF, PENG CH.Research progresses in differentiation of marrow mesenchuymal stem cell to liver cell[J].J Diagn Concepts Pract, 2013, 12 (3) :376-380. (in Chinese) 裴耀飞, 彭承宏.间充质干细胞向肝细胞分化的研究进展[J].诊断学理论与实践, 2013, 12 (3) :376-380. [2]ZHAO JW.Research progresses in treatment of liver fibrosis and cirrhosis using bone marrow stem cells[J].Progr Physiol Sci, 2017, 48 (2) :88-93. (in Chinese) 赵经文.用骨髓干细胞治疗肝硬化研究进展[J].生理科学进展, 2017, 48 (2) :88-93. [3]ZHANG QQ, HANG HL, XIA Q.Status and progresses in treatment of liver failure using stem cells[J].Chin Hepatol, 2015, 20 (6) :483-485. (in Chinese) 张祺祺, 杭化莲, 夏强.干细胞治疗肝衰竭的现状和进展[J].肝脏, 2015, 20 (6) :483-485. [4]WANG DD, CHEN G, HU ZQ, et al.Research advances in mesenchymal stem cells from various sources in treatment of liver fibrosis[J].J Clin Hepatol, 2016, 32 (3) :597-600. (in Chinese) 王冬冬, 陈刚, 胡宗强, 等.不同来源间充质干细胞治疗肝纤维化的研究进展[J].临床肝胆病杂志, 2016, 32 (3) :597-600. [5]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. [6]XU B, WU LL, YANG XM, et al.Treatment of liver injury by subcapsule transplantation of bone marrow mesenchuymal stem cells guided by B ultrasound in rats[J].Chin Hepatol, 2014, 19 (8) :604-606. (in Chinese) 徐兵, 吴林岚, 杨晓梅, 等.B型超声引导下肝包膜下骨髓间充质干细胞移植治疗大鼠肝损伤[J].肝脏, 2014, 19 (8) :604-606. [7]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. [8]XIONG Q, FENG J, WANG J, et al.Comparison among curative effects of three different transplantation approaches of mesenchymal stem cells on rat model of cirrhosis[J].J Third Milit Med Univ, 2011, 33 (8) :804-808. (in Chinese) 熊全, 冯吉, 王军, 等.三种不同途径移植骨髓间充质干细胞治疗肝硬化模型大鼠的效果比较[J].第三军医大学学报, 2011, 33 (8) :804-808. [9]ZHAN YT, CHEN HS, LIU B, et al.Differentiation of rat bone marrow stem cells in liver regeneration[J].J Chin Hepatol, 2006, 22 (2) :121-122. (in Chinese) 展玉涛, 陈红松, 刘宾, 等.骨髓干细胞在大鼠肝再生环境中的分化[J].临床肝胆病杂志, 2006, 22 (2) :121-122. [10]SHEN TG, LI Q, LI J, et al.The influence of fibrin glue on the adhesiveness of human mesenchymal stem cells[J].Chin J Anat Clin, 2009, 14 (3) :171-173. (in Chinese) 申屠刚, 李强, 李骥, 等.纤维蛋白胶对骨髓间充质干细胞黏附作用的影响[J].中华解剖与临床杂志, 2009, 14 (3) :171-173. [11]XIE FR, LIN CB, YAN DX, et al.Fibrin glue combined with bone marrow mesenchymal stem cells to repair multi-layer soft tissue defects in rabbits[J].Chin J Tissue Engineer Res, 2011, 15 (47) :8773-8776. (in Chinese) 谢富荣, 林春博, 严冬雪, 等.生物蛋白胶复合骨髓间充质干细胞修复兔多层软组织缺损[J].中国组织工程研究, 2011, 15 (47) :8773-8776. [12]WU ZM.Preparation and clinic application of cryoprecipitate[J].Chin J Trauma Disabil Med, 2012, 20 (4) :86, 124. (in Chinese) 吴志民.冷沉淀的制备和临床应用[J].中国伤残医学, 2012, 20 (4) :86, 124. [13]JIANG YH.Quality control of preparation process of cold precipitation and its clinical application effect[J].Syst Med, 2016, 1 (11) :22-24. (in Chinese) 蒋永红.冷沉淀制备过程质控及其临床应用效果探讨[J].系统医学, 2016, 1 (11) :22-24. [14]TAN P, TAI JQ, ZHAO LZ.Application of cryoprecipitate in liver transplantation operation[J].J Clinic Transfus Lab Med, 2010, 12 (1) :52-53. (in Chinese) 谭萍, 太继琼, 赵丽芝.冷沉淀在肝移植手术中的应用[J].临床输血与检验, 2010, 12 (1) :52-53. [15]GUO HD, TAN YZ, SHAO SJ, et al.Effects of fibrin glue on prolifiration, migration ischeminc hypoxia protection of marrow derived cardiac stem cell[J].Chin J Integr Med Cardio/Cerebrovasc Dis, 2012, 10 (8) :961-964. (in Chinese) 国海东, 谭玉珍, 邵水金, 等.纤维蛋白胶对骨髓源性心肌干细胞增殖、迁移和缺血缺氧后的保护作用[J].中西医结合心脑血管病杂志, 2012, 10 (8) :961-964. [16]XU B, HUANG SQ, WU LL, et al.Preliminary study on saltingout and frozen-thaw method for exracting cryoprecipitate from plasma in hepatitis patients[J].Chin Hepatol, 2017, 22 (11) :1005-1007. (in Chinese) 徐兵, 黄素钦, 吴林岚, 等.盐析冻融法提取肝炎患者血浆冷沉淀的初步研究[J].肝脏, 2017, 22 (11) :1005-1007. [17]WANG YQ, YANG YJ, GONG JC, et al.Quality analysis of four kinds of cryoprecipitate prepared with different materials[J].J Clin Hematol, 2016, 29 (12) :972-974. (in Chinese) 王艳清, 杨颜菁, 龚嘉聪, 等.4种原料血浆制备的冷沉淀质量分析[J].临床血液学杂志, 2016, 29 (12) :972-974. [18]CHEN JF, PENG J, ZHANG L, et al.An in vitro study on biocompatibility of bioprotein glue used as carrier for BMSCs transplantation[J].Orthopedic J China, 2012, 20 (15) :1424-1427. (in Chinese) 陈继凤, 彭江, 张莉, 等.生物蛋白胶作为骨髓间充质干细胞移植载体的体外相容性研究[J].中国矫形外科杂志, 2012, 20 (15) :1424-1427.
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