摘要
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,10.0μmol.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10.0μmol.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2.5μmol.L-1、5.0μmol.L-1和10.0μmol.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.-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 α1(I).RESULTS: Stimulation with platelet-derived growth factor(PDGF) induced the increase in type I collagen synthesis,while treatment with sorafenib(10.0 μmol/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 -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 μmol/L for 12 h,24 h and 48 h.Sorafenib dose-dependently blocked the mRNA expression of collagen α1(I) in LX-2 cells stimulated with PDGF.Sorafenib at the concentrations of 2.5 μmol/L,5.0 μmol/L and 10.0 μmol/L down-regulated the mRNA expression of collagen α1(I) in LX-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.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2012年第1期85-89,共5页
Chinese Journal of Pathophysiology
基金
河北省自然科学基金资助项目(No.C2007000843
No.C2010000530)
关键词
肝纤维化
肝星状细胞
索拉非尼
胶原
Ⅰ型
血小板源性生长因子
Liver fibrosis
Hepatic stellate cells
Sorafenib
Collagen type Ⅰ
Platelet-derived growth factor