腺病毒介导的心房利钠肽基因在大鼠肺血管平滑肌细胞中的转移导致细胞凋亡。

I. Déprez, M. Darmon, M. Hira, M. Adam, S. Sanquer, E. Teiger, V. Chetboul, M. Eloit, S. Adnot, I. Pham
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引用次数: 3

摘要

心房利钠肽(ANP)对血管平滑肌细胞(VSMCs)具有松弛和抑制生长的作用。为了获得ANP基因在VSMCs中的表达,我们构建了由腺病毒主要晚期启动子(AdMLP-ANP)控制的含有ANP cDNA的重组腺病毒。AdMLP-ANP在5 ~ 100 TCID /细胞的感染倍数范围内治疗肺VSMC后,2天后在细胞培养基中检测到免疫反应性ANP,水平达到101 +/- 27 pmol/well。新表达的ANP具有生物活性,其能够诱导环鸟苷单磷酸在靶细胞中积累,并模仿外源性ANP(10(-8)至10(-7)mol/L)的作用。admlp - anp感染细胞的细胞生长和存活降低,并与VSMC凋亡的促进有关。这些效应发生在10至100 TCID(50)/细胞的多重感染下,在用对照腺病毒构建体(AdMLP-betaGAL和AdMLP-gD)感染的细胞和用外源性ANP(10(-7)至10(-6)mol/L)处理的细胞中都没有观察到。这些结果表明VSMC凋亡对ANP基因表达的响应可能对基因治疗预防血管重塑具有重要意义。
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Adenovirus-mediated transfer of the atrial natriuretic peptide gene in rat pulmonary vascular smooth muscle cells leads to apoptosis.
Atrial natriuretic peptide (ANP) exhibits relaxant and growth-inhibiting effects on vascular smooth muscle cells (VSMCs). To obtain ANP gene expression in VSMCs, we built a recombinant adenovirus containing the ANP cDNA controlled by the adenovirus major late promotor (AdMLP-ANP). After pulmonary VSMC treatment with AdMLP-ANP at a multiplicity of infection ranging from 5 to 100 TCID(50)/cell, immunoreactive ANP was detectable in the cell culture medium at a level that reached 101 +/- 27 pmol/well after 2 days. The newly expressed ANP was biologically active, as evidenced by its ability to induce cyclic guanosine monophosphate accumulation in target cells and to mimic the effect of exogenous ANP (10(-8) to 10(-7) mol/L). Cell growth and survival of AdMLP-ANP-infected cells were decreased and were associated with the promotion of VSMC apoptosis. These effects, which occurred at a multiplicity of infection of 10 to 100 TCID(50)/cell, were observed neither in cells infected with the control adenoviral constructs (AdMLP-betaGAL and AdMLP-gD) nor in cells treated with exogenous ANP (10(-7) to 10(-6) mol/L). These results showing VSMC apoptosis in response to ANP gene expression may have important implications for the prevention of vascular remodeling by gene therapy.
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