骨髓间充质干细胞在大鼠肝纤维化模型中的作用

Effect of bone marrow mesenchymal stem cells on liver fibrosis of rats

目的观察骨髓间充质干细胞（BMSCs）在大鼠肝纤维化模型中（HF）对肝纤维化发展的影响。方法选取雌性Wistar大鼠20只作为研究对象，并随机分成间充质干细胞干预组、单纯造模组各10只。干细胞干预组和单纯造模组以含0．01％浓度的二乙基亚硝胺（DEN）饮水为纤维化诱导剂，干细胞干预组在造模过程中第4、8、12周时经尾静脉输注1×10^6个雄性大鼠BMSCs，单纯造模组经尾静脉注射等量生理盐水作对照。大鼠在同样条件下饲养21周观察下列指标：（1）苏木素．伊红（HE）染色观察肝脏组织的病理形态学变化；（2）原位杂交技术检测实验组大鼠肝脏组织中性别决定基因Y（srY），观察BMSCs移植是否成功；（3）VG特殊染色和免疫组织化学技术检测肝脏组织中的仪．平滑肌肌动蛋白（Ot—SMA），观察BMSCs在肝纤维化模型中的作用。结果病理证实单纯造模组有7只大鼠形成肝纤维化，干细胞干预组有9只大鼠形成肝纤维化；干细胞干预组大鼠肝脏检测到携带SrY的阳性细胞；干细胞干预组大鼠α-SMA的平均阳性表达率为（21．97±11．00）％，高于单纯造模（10．30±3．36）％，两组差异有统计学意义（t=2．69，P〈0．05）；VG特殊染色中，于细胞干预组的纤维化程度（20．63±10．02）％明显高于单纯造模组（11．24±3．S0）％，两组差异有统计学意义（t=2．34，P〈0．05）。结论在DEN制备的大鼠肝纤维化模型中，BMSCs可促进肝纤维化的发展。

Objective To observe bone mesenchymal stem cells （BMSCs） influence on the devel- opment of liver fibrosis. Methods We choose 20 female Wistar rats which randomly divided into interven- tion group （ 10 rats） and control group （ 10 rats）. Interventional group and control group were accepted water which involve consistency as 0.01% diethylnitrosamine. In the 4, 8, 12 week we transplant 1 × 10^6 BMSCs suspension of male rat to interventional group through caudal vein. Control group were injected equal normal saline through caudal vein. The rats were breed in same condition for 21 weeks and observed the following index ： （ 1 ） By the hematoxylin and eosin （HE） staining, observe the pathomorphological change of liver. （2） Detect the existence of SrY by hybridization in situ to confirm successful migrate of BMSCs. （3） VG special coloration and detect the expression level of or-smooth muscle actin （a-SMA） by the immunohistochemistry to observe the differences of two groups. Results There are 7 rats in control group and 9 rats in interventional group which have developed liver fibrosis by HE staining; we discover the existence of SrY in the liver of interventional group, the positive expression rate of ot-SMA in the interven- tional group was （21.97 ＋ 11.00）% ; higher than control group which was （10. 30 ± 3. 36）%, there is significant difference （ t = 2. 69, P 〈 0. 05 ）. The fibrosis in the interventional group （20. 63 ±10. 02） % is apparently higher than control group （ 11.24 ± 3.80 ） %, there is significant difference （ F = 2. 34, P 〈 0.05）. Conclusion In the rat model of liver fibrosis, BMSCs has function of promote to liver fibrosis of rat.