鳖甲煎丸含药血清诱导大鼠骨髓间充质干细胞转化为肝细胞的实验研究

Research on the Transformation of Hepatocyte from Bone Marrow Mesenchymal Stem Cells by Biejiajianwan(鳖甲煎丸) Drug Serum

目的:探讨鳖甲煎丸含药血清对大鼠骨髓间质干细胞(BMSC)向肝细胞分化的影响。方法:体外分离培养大鼠BMSC,取第3代BMSC随机分为空白对照组、肝细胞生长因子联合成纤维生长因子-4(HGF加FGF-4)组、鳖甲煎丸组、HGF加FGF-4+鳖甲煎丸联合组,分别于诱导第7、14、21、28天免疫细胞化学法检测肝细胞标志物甲AFP、ALB及CK18阳性细胞率。结果:各组AFP阳性细胞率均于21 d达到峰值,28 d表达逐渐下降,各组ALB、CK18阳性细胞率随着时间的延长,其阳性比率逐渐升高;在相同时间点,各给药组的ALB、AFP、CK18的阳性细胞率明显高于空白对照组,差异有统计学意义(P<0.05或P<0.01);在第14天至第28天,鳖甲煎丸组、HGF加FGF-4+鳖甲煎丸联合组AFP、CK18阳性细胞率高于HGF加FGF-4组(P<0.05或P<0.01);鳖甲煎丸组在诱导后21、28 d的ALB阳性细胞率明显高于HGF加FGF-4组(P<0.05或P<0.01),而HGF加FGF-4+鳖甲煎丸联合组在诱导后第14天开始,其ALB阳性细胞率均高于HGF加FGF-4组(P<0.05或P<0.01)。结论:鳖甲煎丸含药血清可提高BMSCs向肝细胞定向分化的能力。

Objective: To investigate the effect of Biejiajianwan drug serum on rat bone marrow mesenchymal stem cells to differentiate into liver cells. Methods: The third generation BMSC was isolated and culture in vitro and the third generation BMSC was randomly divided into blank control group, HGF plus FGF-4 group,Biejiajianwan group and HGF plus FGF-4 combined with Biejiajianwan group. Then immunocytochemistry was used to detect the expression of liver cell markers(AFP ALB and CK18) in 7, 14, 21 and 28 days. Results:The positive rate of AFP positive cells reached the peak in each group in 21 days, and the expression rate of ALB and CK18 positive cells increased with the prolongation of time. All medication groups showed higher ALB and CK18 positive rate than blank control group at the same time point(P<0.05 or P<0.01). Biejiajianwan group and HGF plus FGF-4 combined with Biejiajianwan group showed higher positive rate of AFP and CK18 than HGF plus FGF-group at 14 thday and 28 thday(P<0.05 or P<0.01). Biejiajianwan group showed higher rate of ALB positive cells than HGF plus FGF-4 group in 21 days and 28 days after induction(P<0.05 or P<0.01),and HGF plus FGF-4 combined with Biejiajianwan group showed higher ALB positive cells than HGF plus FGF-4 group(P<0.05 or P<0.01) since the 14 thday. Conclusion: The serum of Biejiajianwan can improve the ability of BMSC to differentiate into hepatocytes.