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

详细信息

中图分类号: R575.2
计量
- 文章访问数: 2286
- HTML全文浏览量: 12
- PDF下载量: 611
- 被引次数: 0
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

HTML全文

参考文献(11)

[1]Urtasun R, Lopategi A, George J, et al.Osteopontin an oxi-dant stress sensitive cytokine, up-regulates collagen-I viaintegrinα (V) β (3) engagement and PI3K/pAkt/NFκB signa-ling[J].Hepatology, 2012, 55 (2) :594-608.

[2]Yang J, Zheng J, Wu L, et al.NDRG2 ameliorates hepaticfibrosis by inhibiting the TGF-β1/Smad pathway and alteringthe MMP2/TIMP2 ratio in rats[J].PLoS One, 2011, 6 (11) :e27710.

[3]Fiorucci S, Rizzo G, Antonelli E, et al.Cross-talk betweenfarnesoid-X-receptor (FXR) and peroxisome proliferator-activated receptorγcontributes to the antifibrotic activity ofFXR ligands in rodent models of liver cirrhosis[J].JPET, 2005, 315 (1) :58-68.

[4]Yoshida K, Matsuzaki K.Differential regulation of TGF-β/Smad signaling in hepatic stellate cells between acute andchronic liver injuries[J].Front Physiol, 2012, 3:53.

[5]Jin XZ, Chen YP, Chen Y, et al.The study of Co-culturingCD4+CD25+CD127low/-T cell from patients with HBV-relat-ed liver fibrosis and hepatic stellate cells in vitro[J].J MedRes, 2011, 40 (8) :70-73. (in Chinese) 金晓芝, 陈永平, 程瑗, 等.慢性乙型肝炎肝纤维化患者外周血CD4+CD25+CD127low/-T细胞与人肝星状细胞共培养的实验研究[J].医学研究杂志, 2011, 40 (8) :70-73.

[6]Rath T, Menendez KM, Kügler M, et al.TIMP-1/-2 andtransient elastography allow non invasive diagnosis of cysticfibrosis associated liver disease[J].Dig Liver Dis, 2012, 44 (9) :780-787.

[7]Yang CQ, Hu GL, Tan DM, et al.Relativity of expression ofMMP-1、TIMP-1and variability of typeⅠ、Ⅲcollagen dur-ing experimental liver fibrosis[J].J Clin Hepatol, 2000, 6 (4) :222-224. (in Chinese) 杨长青, 胡国龄, 谭德明, 等.实验性肝纤维化时MMP-1、TIMP-1的表达与Ⅰ、Ⅲ型胶原含量变化的关系[J].临床肝胆病杂志, 2000, 6 (4) :222-224.

[8]Zhang YF, Nie QH, Xu H, et al.Effect of TIMP-2 targetingantisense oligonucleotide on liver function and fibrosis profilesin rat model of liver fibrosis[J].Chin Hepatol, 2005, 10 (4) :291-293. (in Chinese) 张亚飞, 聂青和, 徐辉, 等.以TIMP一2为靶基因的反义寡核苷酸对肝纤维化大鼠肝功能生化和肝纤维化指标的影响[J].肝脏, 2005, 10 (4) :291-293.

[9]Lee J, Seok S, Yu P, et al.Genomic analysis of hepatic far-nesoid X receptor binding sites reveals altered binding in obe-sity and direct gene repression by farnesoid X receptor inmice[J].Hepatology, 2012, 56 (1) :108-117.

[10]Fiorucci S, Rizzo G, Antonelli E, et al.A farnesoid x receptor-small heterodimer partner regulatory cascade modulatestissue metalloproteinase inhibitor-1 and matrix metallopro-tease expression in hepatic stellate cells and promotes reso-lution of liver fibrosis[J].J Pharmacol Exp Ther, 2005, 314 (2) :584-595.

[11]Chen CL, Zhen Z, Liu XP, et al.Relationship between fibro-sis and MMP-2/TIMP-1 in liver tissues from patients withchronic hepatitis B[J].J Clin Hepatol, 2006, 22 (6) :419-421. (in Chinese) 陈超丽, 甄真, 刘晓平, 等.慢性乙型肝炎肝组织中MMP-2、TIMP-l与肝纤维化关系的研究[J].临床肝胆病杂志, 2006, 22 (6) :419-421.

施引文献

资源附件(0)

访问统计