阿魏酸钠对肝纤维化大鼠脂肪干细胞移植存活率及治疗作用的影响

Influence of sodium ferulate on survival rate of adipose-derived stem cell transplantation and therapeutic effect in mice with hepatic fibrosis

目的观察阿魏酸钠对肝纤维化大鼠脂肪干细胞(ASCs)移植存活率及治疗作用的影响。方法选取小鼠50只,予CCl4腹腔注射8周制作肝纤维化模型,将造模成功的40只随机分为A、B、C、D组各10只。A组生理盐水灌胃并经尾静脉注射细胞培养基,B、D组予阿魏酸钠100 mg/kg灌胃1次/d,C、D组经尾静脉行绿色荧光蛋白标记的ASCs移植5×105/只,B、C组分别给予尾静脉注射细胞培养基、生理盐水灌胃,另取10只(E组)不造模且不做任何处理。4周后,检测血清肝功能指标,行Masson染色观察肝组织病理,免疫组化法检测肝组织中的转化生长因子β1(TGF-β1)、α肌动蛋白(α-SMA),荧光显微镜下计数肝组织内存活ASCs数量。结果血清ALT、AST水平A组>B、C、D组>E组,ALB水平E组>A^D组、B组>A组,P均<0.05;A组胶原沉积最多,B、C组较多,D组次之,E组最少;肝组织TGF-β1和α-SMA表达量A组>B组>C组>D组>E组,P均<0.05;C、D组ASCs存活数量比较,P>0.05。结论 ASCs移植联合阿魏酸钠不能明显增加肝内ASCs存活数量,但较单一治疗肝纤维化效果更好。

Objective To observe the influence of sodium ferulate on the survival rate of the transplanted adipose tissue-derived stem cells（ ASCs） and the therapeutic effect in mice with hepatic fibrosis. Methods Fifty mice were selected to make the liver fibrosis models through CCl4 injection intraperitoneally for 8 weeks,and then 40 mice with hepatic fibrosis were randomly divided into groups A,B,C and D. Mice in the group A were administered with saline daily by gavage and then injected with cell culture medium by tail vein. Mice in the groups B and D were given 100 mg / kg sodium ferulate by lavage. Mice in the groups C and D received the GFP labeled ASCs（ 5 × 105） injection. The groups B and D were injected to the caudal veins of mice with cell culture medium and were also gavaged with normal saline. Ten mice（ group E） were selected as the control group without any treatment. After 4 weeks,the liver function index was detected,the pathology of liver tissues was observed by Masson staining,the expression of transforming growth factor-β1（ TGF-β1）,α-smooth muscle actin（ α-SMA） in the liver tissues was determined by immunohistochemistry,and the number of ASCs in liver were detected by fluorescence microscope. Results ALT and AST serum levels： group A groups B,C and D group E（ P 〈0. 05）. ALB serum levels： group E groups A,B,C and D,group B group A（ P 〈0. 05）. Collagen deposition：group A groups B and C group D group E. Expression of TGF-β1and α-SMA： group A group B group C group D group E（ P 〈0. 05）. There was no significant difference in survival rate between group C and group D（ P 〈0. 05）. Conclusion The combined application of sodium ferulate and ASCs can not significantly increase the survival rate of transplanted ASCs in liver,but has a better therapeutic effect as compared with the separated application.