【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(P值均>0.05)。体外培养4 d后,大鼠FG-MSCs的细胞增殖率均略高于单纯MSCs,但差异均无统计学意义(P值均>0.05)。肝损伤大鼠肝组织内注射FG-MSCs或MSCs治疗后,大鼠血浆ALT、TBil浓度明显降低(P值均<0.05),Alb浓度均明显增高(P值均<0.05)。结论将FG-MSCs或MSCs注入肝实质组织内均可明显改善肝损伤大鼠的肝功能,FG-MSCs兼可防止注射出血。更多还原

【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.更多还原