中文摘要：

目的探讨改变组蛋白乙酰化修饰水平对转化生长因子β1（TGF-β1）调控系膜细胞（MC）内纤溶酶原激活物抑制物1（PAI-1）基因表达的影响。方法利用细胞转染技术使MC内高表达组蛋白乙酰化酶（HAT）的CREB结合蛋白（CBP）、P300或去乙酰化酶（HDAC）1、3、5，通过荧光素酶检测、实时定量PCR和Western印迹法，观察TGF-β1（5tzg／L）刺激下MC内PAI-1基因转录活性、mRNA及蛋白水平的改变；并利用组蛋白去乙酰化酶抑制剂曲古抑菌素A（TSA），观察其对PAI-I基因表达及TGF-β1-Smad蛋白信号途径的活性影响。结果TGF-β1刺激增加了PAI-1基因的转录活性和mRNA水平（P〈0．05）；高表达CBP和P300显著促进了TGF-β1对PAI-1基因转录活化和mRNA及蛋白表达的调控（P〈0．05）；而高表达HDAC1、3、5及突变型CBP、P300则明显抑制了TGF-β1诱导PAI-1基因转录表达的调节（P〈0．05）。改变细胞内HAT／HDAC水平没有影响TGF-β1诱导Smad2／3蛋白磷酸化的作用。HDAC抑制剂TSA显著增加了TGF-β1诱导PAI-1基因转录表达的调节，但对TGF-β1-Smad2／3信号途径的活化作用没有影响。结论改变细胞内组蛋白乙酰化修饰水平能够影响TGF-β1对PAI-1基因的表达调控。

英文摘要：

Objective To investigate the effect of histone acetylation change on the transforming growth factor -β1 （TGF-β1）-associated plasminogen activator inhibitor 1 （PAI-1） regulation in mesangial cells （MCs）. Methods MCs were transfected with histone acetyltransferase （HAT） CREB-binding protein（CBP）, P300 or histone deacetylase （HDAC） 1, 3, 5 expression vectors, followed by luciferase assay, real- time PCR and Western blotting to see the change of PAI- 1 gene＇s transcriptional activity, mRNA and protein in response to TGF-β1. HDAC inhibitor trichostatin A（TSA） was used and TGF-β1-Smad signaling activity was detected also in this experiment. Results TGF-β1 enhanced PAI- 1 transcriptional activity and mRNA expression （P 〈 0.05）. Over- expression of CBP or P300 significantly increased TGF-β1- associated PAI- 1 transcriptional activity, mRNA and protein expression （P 〈 0.05）. On the contrary, over-expression of HDAC1, 3, 5 or dominant negative CBP or dominant negative P300 obviously reduced PAI- 1 gene＇s expression induced by TGF-β1 （P 〈 0.05）.Change of HAT/HDAC did not affect TGF-β1-associated Smad2/3 phosphorylation. TSA enhanced TGF -β1 induced PAl-1 gene＇s regulation markedly, but did not change TGF-β1-Smad2/3 signaling activity. Conclusion Change of histone acetylation can affect TGF-β1 associated PAI- 1 gene＇s regulation in MCs.