Beyea J A, Olson D M, Harvey S
Department of Physiology, University of Alberta, AB, Canada.
Mol Cell Biochem. 2009 Jan;321(1-2):197-204. doi: 10.1007/s11010-008-9933-z. Epub 2008 Nov 5.
Growth hormone (GH) mRNA and protein have recently been demonstrated in the rat lung throughout the period of alveolarization (day 4-14 postnatally). The functional significance of this finding was therefore assessed, by determining the effects of GH mRNA knockout using aerosolized antisense oligodeoxynucleotides (ODN) directed against the GH gene. In a preliminary experiment, the effectiveness of the antisense GH ODN was demonstrated in a lung Type II epithelial cell line (L2 cells), in which constitutive GH mRNA expression was completely abolished by GH ODN transfection. Administration of the aerosolized GH ODN to 4-day-old rats for 10 days was accompanied by a widespread presence of its delivery liposomes within lung cells. Aerosolized GH ODN treatment decreased lung concentrations of IGF (insulin-like growth factor)-1 and increased concentrations of albumin, calcyclin binding protein, superoxide dismutase, RNA binding protein motif 3, and the alpha- and beta-subunits of ATP synthase and electron transfer flavoprotein. At least 32 other proteins (identified by 2D gel electrophoresis) were also significantly affected by the antisense GH ODN treatment. By changing the lung proteome, these results indicate hitherto unsuspected autocrine/paracrine actions of GH in developmental lung function.
最近已证实在大鼠肺泡形成期(出生后第4 - 14天)整个阶段,肺中均有生长激素(GH)mRNA和蛋白质存在。因此,通过使用针对GH基因的雾化反义寡脱氧核苷酸(ODN)来确定GH mRNA敲除的影响,从而评估这一发现的功能意义。在一项初步实验中,反义GH ODN在肺II型上皮细胞系(L2细胞)中显示出有效性,通过GH ODN转染可完全消除其组成型GH mRNA表达。给4日龄大鼠雾化给予GH ODN 10天,其递送脂质体广泛存在于肺细胞中。雾化GH ODN治疗降低了肺中胰岛素样生长因子(IGF)-1的浓度,并增加了白蛋白、钙结合蛋白、超氧化物歧化酶、RNA结合蛋白基序3以及ATP合酶和电子传递黄素蛋白的α和β亚基的浓度。至少32种其他蛋白质(通过二维凝胶电泳鉴定)也受到反义GH ODN治疗的显著影响。通过改变肺蛋白质组,这些结果表明GH在发育中的肺功能中存在迄今未被怀疑的自分泌/旁分泌作用。