不同途径移植间充质干细胞对大鼠急性肝损伤的修复作用

Repair effect of mensenchymal stem cells transplantation by different routes on acute liver injury of rats

目的:评价骨髓间充质干细胞(MSCs)移植对大鼠急性肝损伤的修复作用,探讨其可行性和有效性。方法:四氯化碳(CCl4)腹腔注射复制Wistar大鼠急性肝损伤模型,造模后随机分为实验组及对照组,实验组分别通过尾静脉、门静脉、肝内注射移植经Hochest33342标记的大鼠骨髓MSCs悬液0.5mL(约1×106个细胞),对照组分别经相同途径注射等量的PBS。移植后3和7d检测血清生化指标,观察Hochest33342标记细胞的定位及肝脏组织病理学变化。结果:细胞移植术后3和7d,尾静脉注射组丙氨酸氨基转移酶(ALT)及天冬氨酸氨基转移酶(AST)水平与对照组比较差异无统计学意义(P>0.05),门静脉、肝内注射组与相应的对照组比较显著下降(P<0.01),肝内注射组与门静脉组比较显著下降(P<0.05)。尾静脉组各时间点肝脏内均未见Hochest33342标记细胞;门静脉组3d时肝脏内可见散在亮蓝色荧光的细胞,数量较少,7d时数量稍多;肝内注射组3及7d肝脏内均可见较多蓝色荧光的细胞;对照组各时间点均未见Hochest33342标记细胞。大鼠CCl4诱导急性肝损伤24h后肝组织结构紊乱,肝小叶中央区呈弥漫性多发性片状坏死,肝细胞呈弥漫性脂肪变性,汇管区及中央静脉附近炎性细胞浸润明显。尾静脉注射组各时间点与对照组比较无明显形态学改变;门静脉、肝内注射组3d时大鼠肝小叶中央区仍有小部分肝细胞坏死,肝细胞脂肪变性、炎性细胞浸润等,但较相应对照组有改善;门静脉、肝内注射组7d时肝细胞坏死、充血、炎性细胞浸润继续好转,肝内注射组病理组织学改善情况较门静脉组更明显。结论:MSCs经门静脉、肝内注射移植于急性肝损伤大鼠体内,明显改善了受损肝脏的功能,对急性肝损伤具有修复作用,肝内注射途径移植优于门静脉途径。

Objective To evaluate the repair effect of bone marrow mensenchymal stem cells（MSCs） transplantation on acute liver injury of rats and to explore the feasibility and validity.Methods The Wistar rats were treated with 10% CCl4 olive oil mixture by intraperitoneal injection to copy acute liver injury model.The model rats were randomly divided into treatment and control groups.Hochest33342-labelled rat bone marrow MSCs 0.5 mL（1×106 cells） were transplanted into the rats by tail vein,portal vein and liver injection in treament groups,respectively.The same volume PBS was transfused into the rats in various control groups.On day 3 and 7 after transplantation,the liver function indexes of rats were detected,the distribution of Hochest33342-labelled cells and the pathological changes of liver tissue were observed by fluorescence microscopy at same time.Results Compared with control group,the levels of serum ALT and AST were decreased on day 3 and day 7 after transplantation in intraportal vein and intraliver injection groups（P〈0.01）,but there were no statistically significant differences in tail vein group（P〉0.05）;and they were decreased in intraliver injection group compared with intraportal vein group（P〈0.05）.The Hochest33342 labelled cells were not found in vein group and control groups at different time,but they were found in intraportal vein and intraliver injection groups;the days were more,the cells were more.The pathological structure of liver displayed multiple necrosis of hepatocyte,diffuse degeneration of hepatocyte and infiltration of inflammatory cells in rats treated with CCl4 for 24h.The severity of pathological structure of liver wasn’t alleviated in tail vein group at different time after transplantation,but it was alleviated in intraportal vein and intraliver injection groups on day 3 and day 7;the improvement of pathohistology intraliver injection group was better than that in intraportal vein group.Conclusion Intraportal vein,intraliver injection transplantation of bone marrow MSCs can ameliorate and repair liver function of rats with acute liver injury.The route of intraliver injection transplantation is better than intraportal vein transplantation.