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Effect of drug-containing serum of Si-Ni-San on inducing differentiation of rat′s hepatic stem cells and its regulatory mechanism |
XU Wei-dong1, QIAN Yuan-xia1, GAO Jing1, ZHAO Ying-qian2 |
(1. School of Pharmacy, Jiangsu University, Zhenjiang Jiangsu 212013; 2. Clinical School of Traditional Chinese Medicine, Hubei University of TCM, Wuhan Hubei 430065, China) |
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Abstract Objective: To study the effect of drug-containing serum of Si-Ni-San on differentiation of rat′s hepatic stem cells and its mechanism, and explore the scientific basis for treatment of liver disease. Methods: HPLC-MS-MS method was used to detect drugcontaining serum of Si-Ni-San; WB-F344 cells were treated with drugcontaining serum, the activity of cells was detected by CCK-8 assay, the glucose consumption was assessed by glucometer, generation of lactic acid was determined by LA kit, ATP content was measured by luciferin luciferase method,and the expressions of epidermal growth factor receptor(EGFR), fibroblast growth factor receptor(FGFR),transforming growth factor beta receptor type I(TGF-βRⅠ),hepatocyte growth factor receptor(HGFR) were observed by western blotting. Results: In drug-containing serum of Si-Ni-San, some ingredients in prototype like paeoniflorin and aurantiamarin, others in metabolite type such as naringenin- glucuronic acid and glycyrrhetinic acid. Different concentrations of drug-containing serum showed no toxic effects on WB-F344 cells after been incubated with drug-containing serum, 10% concentration of drug-containing serum could significantly promote proliferation of WB-F344 cells. The levels of glucose consumption and lactic acid decreased with the time of differentiation, but ATP took on a up-regulating trend. the expressions of EGFR, FGFR, TGFβRI and HGFR also increased as drug-containing serum incubation. Conclusion: The Si-Ni-San containing serum could promote hepatic stem cells′ proliferation and induce their differentiation, and maybe the regulatory mechanism is related to cytokine receptors, such as EGFR,HGFR.
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Received: 11 June 2014
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