过氧化物酶体增殖物激活受体γ1基因转染诱导肝星状细胞表型及功能的变化  

Effects of lentiviral vectors-mediated overexpression of PPARγ1 on the biochemical features of hepatic stellate cells

作　　者：熊伍军[1] 刘菲[1] 何益[1] 江鸣[1] 刘雁冰[1] 赵中辛[2] 

机构地区：[1]同济大学附属东方医院消化科,上海200120 [2]同济大学附属东方医院普外科,上海200120

出　　处：《中华实验外科杂志》2008年第11期1412-1414,共3页Chinese Journal of Experimental Surgery

基　　金：上海市青年科技启明星计划资助项目（05QMX1444）

摘　　要：目的用慢病毒介导PPARγ1基因在HSC中过表达，观察对HSC的增殖及细胞外基质合成的影响。方法用慢病毒质粒系统在293T细胞中包装出重组慢病毒Lent／PPARγ1并感染HSC，用RT—PCR及WB检测目的基因及目的蛋白的表达，用噻唑蓝（MTT）比色法观察对HSC增殖的影响，用RT-PCR观察对Ⅰ型胶原、TGFβ1表达的影响。结果成功构建Lent／PPARγ1并感染HSC，RT-PCR及WB检测到目的基因及蛋白的表达，MTT检测表明Lent／PPAR3,1组12、96h的A值分别为0．420±0．031、0．638±0．040，与对照组0．448±0．019、0．810±0．070比较差异有统计学意义（P〈0．05）；RT—PCR检测表明Lent／PPARTI组Ⅰ型胶原、TGFβ1的2^△△Ct值分别为1．496±0．359、0．667±0．153，与对照组2．602±0．301、1．008±0．102比较，差异有统计学意义（P〈0．01）。结论成功构建携带PPARγ1的重组慢病毒载体并感染HSC，PPARγ1基因过表达可抑制HSC的增殖和活化。Objective To study the effects of overexpression of PPARγ1 gene via a lentiviral vector on biochemical features of culture-activated hepatic stellate ceils （HSC）. Methods Lent/PPARγ1 was constructed from 293T cells and transfected into HSC. RT-PCR and Western blot were performed to detect the expression of PPARγ1. The effects of Lent/PPARγ1 on the proliferation of HSC was examined by using MTT. The expression of collagen type Ⅰ and TGFβ1 were examined by using RT-PCR. Results Lent/PPARγ1 was constructed and transfected into HSC successfully. RT-PCR and Western blot detected the expression of the aimed gene and protein. MTT showed in the Lent/PPARγ1 group the A values of 12 h （0. 420 ± 0.031 ） and 96 h （ 0. 638 ±0. 040 ） were significantly lower than in the control group [ （0.448 ±0.019 ） and （ 0.810± 0. 070 ）, respectively ] （ P 〈 0.05 ）. RT-PCR revealed in the Lent/ PPAR＇yl group the 2^△△Ct, values of type Ⅰ collagen [ （ 1. 496 ± 0. 359 ） ] and TGFβ1 [ （ 0. 667 ± 0. 153 ）] were significantly lower than in the control group [ （ 2. 602 ± 0. 301 ） and （ 1. 008 ± 0. 102 ）, respectively ] （P 〈 0. 01 ）. Conclusion Lent/PPARγ1 was constructed and transfected into HSC saccessfully. The overexpression of PPARγ1 could inhibit the proliferation and activation of HSC.

关 键 词：慢病毒 肝星状细胞 基因转染 

分 类 号：R575.2[医药卫生—消化系统]