目的 研究中药复方牛黄、西洋参(简称牛黄参)含药血清对大鼠肝干细胞系WB-F344增殖、分化的影响,探讨益气解毒法体外诱导肝干细胞分化为肝细胞的内在机制.方法 采用中药血清药理学方法,以不同浓度的牛黄参含药血清(5%、10%、20%)作用于WB-F344,以正常大鼠血清(5%、10%、20%)处理的WB-F344为空白对照组,以单纯培养体系处理的WB-F344为正常对照组;噻唑兰(MTT)比色法检测不同浓度含药血清对WB-F344增殖的影响;取增殖的最佳浓度,蛋白印迹法(western blot)检测含药血清对WB-F344细胞HGFR、EGF、EGFR、TGFβ1R、TGFβ2R表达的影响;逆转录-聚合酶链反应(RT-PCR)检测含药血清对WB-F344肝细胞表面标志物白蛋白(ALB)的mRNA表达的影响.结果 牛黄参含药血清能促进WB-F344的增殖(P ＜ 0.01),且10%为增殖的最佳浓度梯度;能刺激WB-F344细胞HGFR、EGF、EGFR、TGF-β1R、TGF-β2R的表达量上调(P ＜ 0.01);能促进WB-F344肝细胞表面标志物ALB的mRNA表达(P ＜ 0.01).结论 牛黄参含药血清可以通过细胞因子体系对肝干细胞WB-F344的增殖、分化进行调节,为利用中药复方制剂调控肝干细胞,促进肝再生和肝损伤修复提供实验与理论基础.%Objective To study the effects of NHS participation drug-containing serum with Chinese herbal compound Taurine and Ginseng on the differentiation and proliferation of rat hepatic oval cell line WB-F344 and explore intrinsic mechanism of the method of tonifyingqi and detoxication on hepatic stem cells differentiate into liver cells. Methods Traditional Chinese medicine serum pharmacology method were used, WB-F344 cells were treated with NHS participation drug-containing serum of different concentrations (5%, 10%, 20%), the blank control group was treated with the normal rat serum of different concentrations (5%, 10%, 20%), and normal control group was treated with simple culture system. MTT colorimetry was used to detect the effects of different concentrations of drug serum on the proliferation of WB-F344; the optimal concentration of proliferation was taken, western blotting was used to detect the effects of drug serum on the expressions of WB-F344 cells'HGFR, EGF, EGFR, TGFβ1R and TGFβ2R; and reverse transcriptase polymerase chain reaction (RT-PCR) method was used to detect the effects of drug serum on the mRNA expression of WB-F344 cells'liver cell surface markers ALB. Results NHS participation drug-containing serum could promote the proliferation of WB-F344 (P < 0.01), 10% for the proliferation of optimal concentration gradient; and could stimulate the rising of expression of WB-F344 cells'HGFR, EGF, EGFR, TGFβ1R and TGFβ2R (P < 0.01), which could promote the mRNA expression of WB-F344 cells'liver cell surface markers ALB (P < 0.01). Conclusion NHS drug serum may regulate and control the proliferation and differentiation of liver stem cell line WB-F344 by cytokine system, which may provide the experimental and theoretical basis for using chinese medicine compound to regulate and control hepatic stem cells and promote liver regeneration and repair of liver injury.
展开▼
