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

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(MIT)比色法观察对HSC增殖的影响,用RT-PCR观察对Ⅰ型胶原、TCFβ1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/PPARγ1组12、96 h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/PPARγ1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.

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

Objective To study the effects of overexpression of PPARγ1 gene via a lentiviral vec-tor on biochemical features of culture-activated hepatic stellate cells (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/ PPARγ1 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 successfully. The overexpression of PPARγ1 could inhibit the proliferation and activation of HSC.