法尼酯衍生物X受体活化对肝星状细胞TIMP-1、TIMP-2及MMP-2表达的调节作用

陈科全; 周碧瑶; 陈雅莹; 邹原方; 周宇

法尼酯衍生物X受体活化对肝星状细胞TIMP-1、TIMP-2及MMP-2表达的调节作用

1. 广州医学院第一附属医院消化内科
2. 中山大学附属东华医院肾内风湿科
3. 广东医学院附属医院消化内科

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中图分类号: R575.2
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出版历程
- 出版日期: 2013-04-20

Regulatory effeots of FXR activationon expression of TIMP-1, TIMP-2 and MMP-2 in hepatic stellate cells

摘要

摘要:
目的研究法尼酯衍生物X受体（FXR）对肝星状细胞基质金属蛋白酶组织抑制因子-1（TIMP-1）和基质金属蛋白酶组织抑制因子-2（TIMP-2）及基质金属蛋白酶-2（MMP-2）表达的影响。方法应用FXR人工合成配体GW4064（0.01、0.1、1μmol/L）处理大鼠肝星状细胞株HSC-T6后,应用实时荧光定量PCR法检测FXR、TIMP-1、TIMP-2及MMP-2 mRNA表达变化,应用Western Blot法检测TIMP-1、TIMP-2及MMP-2蛋白表达的变化。结果 GW4064处理HSC-T6后,FXR mRNA相对表达量明显增加（P=0.000）,TIMP-1及TIMP-2 mRNA及蛋白相对表达量明显减少（P<0.01）,GW4064浓度在1μmol/L时MMP-2mRNA及蛋白相对表达量则明显增加（P=0.000）。结论 GW4064通过激活FXR下调TIMP-1和TIMP-2表达及上调MMP-2的表达来调控细胞外基质的合成和降解的平衡,提示FXR配体可能可以治疗肝纤维化。
- 星形细胞 /
- 肝细胞 /
- 肝硬化 /
- 基质金属蛋白酶类
Abstract:
Objective To determine whether the regulator of bile acid and carbohydrate metabolism in hepatic stellate cells (HSCs) , Farnesoid X receptor (FXR) , mediates the expression of fibrosis-related genes tissue inhibitor of matrix metalloproteinase (TIMP) -1, TIMP-2, and matrix metalloproteinase-2 (MMP-2) .Methods An in vitro cell culture system with the rat HSC-T6 line was used to evaluate the effects of FXR by treating with the synthetic FXR agonist GW4064 at various concentrations (0.01, 0.1 and 1 μmol/L) for 18 h.Untreated cells served as controls.The mRNA levels of FXR, TIMP-1, TIMP-2, and MMP-2 were measured by real-time reverse transcription PCR.The protein levels of TIMP-1, TIMP-2, and MMP-2 were determined by western blotting.The significance of intergroup differences was assessed by single-factor one-way ANOVA statistical analysis.Results Treatment with GW4064 led to significantly increased mRNA expression of FXR (0.01 μmol/L vs.control, P<0.01) .While the 0.1 μmol/L concentration of GW4064 stimulated a significantly higher level of FXR mRNA expression than the 0.01 μmol/L concentration (P<0.01) 1="" the="" level="" was="" not="" significantly="" different="" from="" that="" stimulated="" by="" l="" concentration="" p="">0.05) .Unlike the 0.01 μmol/L concentration of GW4064, the 0.1 and 1 μmol/L concentrations reduced the TIMP-1 and TIMP-2 mRNA and protein expressions to levels significantly lower than that in the controls (all P<0.05) .GW4064 treatment increased MMP-2 mRNA and protein expressions and the 1 μmol/L mediated increase was significantly higher than that of the control (P<0.01) .Conclusion Activation of FXR on HSCs may contribute to fibrosis by down-regulating TIMP-1 and TIMP-2 and up-regulating MMP-2, which mediate the balance of extracellular matrix synthesis and degradation;thus, FXR ligands may represent useful therapeutic targets of liver fibrosis.
- astrocytes /
- hepatocytes /
- liver cirrhosis /
- matrix metalloproteinases

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参考文献(11)

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