不同方式注射骨髓间充质干细胞对溃疡性结肠炎大鼠模型的疗效

Comparison of therapeutic effects in different ways of bone marrow mesenchymal stromal cells injection in rats with ulcerative colitis

目的对比心脏注射和鼠尾静脉注射骨髓间充质干细胞(bone marrow mesenchymal stromal cells,BMSCs)对溃疡性结肠炎(ulcerative colitis,UC)大鼠模型的疗效,并分析可能的治疗机制。方法体外扩增培养雄性SD大鼠的BMSCs,用5%三硝基苯磺酸(trinitrobenzenesulfonic acid,TNBS)/乙醇灌肠建立UC大鼠模型,并建立正常对照组D。造模后24 h,A组由心脏注射BMSCs悬液0.5 ml(含1×106个干细胞),B组由鼠尾静脉注射BMSCs悬液0.5 ml(含1×106个干细胞),C组由鼠尾静脉注射0.5 ml 0.9%氯化钠注射液,计算各组大鼠的疾病活动指数(disease activity index,DAI)。移植后7 d处死大鼠,光镜下观察结肠病理变化,及人性别确定区Y蛋白(human sex-determining region Y protein,SRY蛋白)、自杀相关因子(factor associated suicide,FAS)、G-蛋白偶联受体5(leucine-rich repeat containing g protein coupled receptor 5,Lgr5)的表达。结果 A组较B、C组DAI评分低(F=90.603,P<0.05),但A、B组间差异无统计学意义。干细胞移植组大鼠均检测到SRY蛋白,A组优于B组;C组Fas表达高于D组(t=9.494,P<0.05),A、B组Fas表达较C组下调,且A组低于B组,3组差异有统计学意义(F=38.6,P<0.05);A、B组LGR-5较C组表达明显增加,且A组较B组高,3组差异有统计学意义(F=24.7,P<0.05)。结论心脏注射BMSCs对UC的疗效优于鼠尾静脉注射;BMSCs可通过减轻肠道细胞凋亡、促进肠道干细胞再生起到对UC的治疗作用。

Objective To compare the efficacy of intracardiac injection and intravenous injection of bone marrow mesenchymal stromal cells （BMSCs） in rats with ulcerative colitis （UC） and analyze the therapeutic mechanism.Methods BMSCs from SD male rats were in vitro cultured.The rat model of UC was established by 5％ trinitrobenzenesulfonic acid （TNBS）/ethanol enema.Group D was the control group.And 24 hours later,rats of group A were injected with 0.5 ml of BMSCs suspension via the heart （1 × 106）.Rats of group B were injected with 0.5 ml of BMSCs suspension via tail vein （1 × 106）.Rats of group C were injected with equal volume of normal saline via tail vein.Disease activity index （DA1） of rats in every group was observed and calculated.The rats were sacrificed at day 7 after injection.The pathological changes of the colon and the expression of human sex-determining region Y protein （SRY protein）,factor associated suicide （FAS）,leucine-rich repeat containing g protein coupled receptor 5 （Lgr5） were observed under microscope.Results DAI of group A was lower than that of group B and C （F=90.603,P ＜ 0.05） but there were no differences between group A and B.The repairing of injured colon in group A was much better than in group B.The SRY protein could be detected in transplantation group.Compared with the control group,the expression of Fas in group C increased （t=-9.494,P ＜ 0.05）.Compared with group C,the expression of Fas in treatment group decreased.And the expression in group A was lower than in group B.There were significant difference among the three groups （F=38.6,P ＜ 0.05）.Compared with group C,the expression of LGR-5 in treatment group was significantly increased.And the expression in group A was higher than in group B.There were significant difference among the three groups （F=24.7,P ＜ 0.05）.Conclusion The efficacy of intracardiac injection of BMSCs is much better than the efficacy of intravenous injection in rats with UC.BMSCs plays a therapeutic role in rats with UC by reducing intestinal cells apoptosis and promoting intestinal stem cell regeneration.