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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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