目的:研究索拉非尼(sorafenib)对人肝星状细胞胶原合成的影响.方法:应用人肝星状细胞株LX-2进行体外研究,采用[3H]-脯氨酸掺入法测定胶原的合成,采用免疫细胞化学法检测I型胶原蛋白表达,采用real-time PCR法测定I型胶原α1 mRNA表达.结果:免疫细胞化学研究显示血小板源性生长因子(PDGF)刺激可引起LX-2细胞胶原合成增加,10.0 μmol·L-1索拉非尼作用于LX-2细胞 24 h能明显抑制I型胶原蛋白的合成.无论有无PDGF的刺激,索拉非尼均呈剂量与时间依赖性地抑制LX-2细胞胶原合成(P<0.01);在10.0 μmol·L-1浓度下,索拉非尼作用于LX-2细胞 12 h、24 h和48 h对胶原合成的抑制率为22.69%、37.52%和71.74%.索拉非尼剂量依赖性地抑制PDGF诱导的I型胶原α1 mRNA表达上调;在2.5 μmol·L-1、5.0 μmol·L-1和10.0 μmol·L-1 索拉非尼作用下,I型胶原α1 mRNA表达较PDGF刺激组分别下调58.66%、67.06%和81.64%.结论:索拉非尼在体外能抑制人肝星状细胞胶原的合成,抑制I型胶原的表达,有可能成为一种新型的治疗肝纤维化药物.%AIM:To investigate the effects of sorafenib on collagen synthesis in human hepatic stellate cells ( HSCs ). METHODS : HSC cell line LX - 2 was used in vitro in this study. [3 H ] - proline incorporation assay was performed to measure the collagen synthesis. Immunocytochemistry was applied to detect type I collagen and real -time PCR was used to determine the mRNA expression of collagen al ( I). RESULTS: Stimulation with platelet - derived growth factor ( PDGF ) induced the increase in type I collagen synthesis, while treatment with sorafenib ( 10. 0μmol/L) for 24 h markedly decreased the collagen synthesis. Sorafenib resulted in dose - dependent and time - dependent decrease in collagen synthesis in LX -2 cells in the absence or presence of PDGF by [3 H ] - proline incorporation assay. The inhibition rates were 22. 69% , 37. 52% and 71.74% , respectively, when LX -2 cells was treated with sorafenib at 10.0 (xmol/L for 12 h, 24 h and 48 h. Sorafenib dose - dependency blocked the mRNA expression of collagen al (I) in LX -2 cells stimulated with PDGF. Sorafenib at the concentrations of 2. 5μmol/L, 5. 0 (xmol/L and 10. 0μmol/L down - regulated the mRNA expression of collagen al ( I) inLX-2 cells by 58. 66%, 67.06% and 81. 64%, respectively. CONCLUSION: Sorafenib inhibits the collagen synthesis and blocks the expression of type I collagen at mRNA and protein levels in vitro in LX - 2 cells.Therefore, sorafenib may be a potential therapeutic agent in the treatment of liver fibrosis.
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